Here - OAK

Transcription

Here - OAK

Ninth Annual OAK Meeting
Danish Brain Research Laboratories Meeting
Copenhagen 11-12 June 2010
BRIC - Biotech Research & Innovation Center
University of Copenhagen
Ole Maaløes Vej 5
2200 Copenhagen N
P R O G R A M
Friday 11th of June 2010
10:55
Welcome - Flemming Fryd Johansen
BRIC SEMINAR ROOM 1.2.03
11:00-12:00 SCHIZOPHRENIA
Chair: Bente Finsen
11:00
Celia Kjærby Hansen : Determination of GABAergic functioning in prefrontal cortex of
early postnatal PCP rats
11:15
Jesper Riise: Stereological quantification of parvalbumin positive interneurons in prefrontal cortex of two animal models of schizophrenia
11:30
Mark Holm Christensen : Immunohistochemical investigations of parvalbumin, GAD65
and GAD67 protein levels in a sub-chronic PCP model
11:45
Katrine Fabricius : Social Isolation of rats as a possible animal model of some traits of
schizophrenia
12:00 - 12:45
LUNCH
BRIC LUNDBECK AUDITORIUM
12:45-14:00 PATHOPHYSIOLOGY OF BRAIN INJURY
Chair: Bente Pakkenberg
12:45
Reza Khorooshi : Type I interferons signaling in axonal injury-induced glial response
13:00
Ditte Elmann : The role of neuronal nuclear factor-kappa B after moderate spinal cord
injury
13:15
Nina Eriksen : The influence of parenchymal damage on cortical spreading
13:30
Kim Anker Kristiansen : Expressional change in calcitonin gene-related peptide, receptor activity modifying protein 1 (RAMP1) and bradykinin 1 receptor after experimental inflammation in rat trigeminal ganglia
13:45
Zindy Raida : Application of a transient middle cerebral artery occlusion stroke model
in mice
14:00-14:15
COFEE BREAK
14:15-14:45 NEUROTRANSMISSION I
Chair: Flemming Fryd Johansen
14:15
Anne M Landau : Noradrenaline release: a potential antidepressant mechanism of
brain stimulation therapies
14:30
Casper Gøtzsche : Viral vector-induced overexpression of neuropeptide Y receptors in
treatment of seizures
14:45-15:30
COFFEE BREAK & LAB-TOUR
P R O G R A M
Friday 11th of June 2010
15:30-16:00 STEREOLOGY
Chair:
Arne Møller
15:30
Anna Schou Karlsen: Comparative study of the human brain using histology, stereology and MRI
15:45
Allan Rasmusson : Virtual slides: Development of automatic neuro-analysis tools and
verifying them using stereology
COFFEE BREAK
16:15-17:30 NEUROINFLAMMATION
Chair: Yawei Liu
16:00-16:15
16:15
Mohammed Salem : The effect of Interferon regulatory factor 7 on the development of
experimental autoimmune encephalomyelitis in mice
16:30
Christina Fenger : Transcriptional regulation of myelination/remyelination processes in
the mouse central nervous system
16:45
Christian Berg : Cell Cycle Signaling in Interferon mediated Neuroprotection
17:00
Martin Kuss : Interferon-β in CNS homeostasis and neurodegeneration : A matter of
impaired autophagy
17:15
Jeanette Svensson : CD1-deficient mice develop spontaneous gait ataxia and depressive symptoms
17:30-17:45
COFEE BREAK
17:45-18:30 PLENARY LECTURE by Roger Pocock
Chair: Yawei Liu
17:45
Roger Pocock : Neuronal responses to hypoxic insults - lessons from the worm
18:30-19:30
Hotel check-in & OAK board meeting
20:00-23:00
C o n f e r e n c e G a l l a D i n n e r, R E S TA U R A N T R O M A R I N ,
Ryesgade 90, Tlf. 35432928
P R O G R A M
Saturday 12th of June 2010
09:00-09:45DOPAMINE
Chair: Trevor Owens
09:00
Aage K Olsen Alstrup : Pigs in PET: Differing effects of propofol versus isoflurane on
dopamine D1 receptor binding in Göttingen minipig brain
09:15
Line Gebauer : The story of music and dopamine - a pleasurable surprise
09:30
Gunnar Sørensen : Subtle C-terminus mutation of the dopamine transporter leads to
attenuated psychostimulant response
09:45-10:15
COFEE BREAK
10:15-10:45 NEUROTRANSMISSION II
Chair: Uffe Kristiansen
10:15
Nadia Fredsø Andersen : A prospective study of epileptogenesis in dogs
10:30
Ditte Dencker Nielsen : Role of M4 muscarinic receptors in psychostimulant-mediated
effects
10:45-11:00
COFEE BREAK
11:00-11:45 AMYLOIDOSIS
11:00
Rebecca Radde : Microglial Response in a Mouse Model for Cerebral Amyloidosis
11:15
Laura Ilkjaer : Expression of IL-1β and TNF-α in Alzheimer’s pathology in mice before
and after endotoxin challenge
11:30
Louise T Jensen : Epi-1 - a novel gene regulating amyloid-beta toxicity in a worm
model of Alzheimer's disease
11:45-12:00
COFEE BREAK
12:00-12:15 ANIMAL MODELS
Chair: Albert Gjedde
12:00
Susanne Kaae : Hippocampal neuroplasticity in a genetic rat model of depression
12:15-13.00 PLENARY LECTURE by Albert Gjedde
12:15
Albert Gjedde : Using yohimbine to map alpha-2 adrenoceptor occupancy in pig cerebral cortex in vivo: Theory, method, and preliminary binding potentials
13.00
Farewell by OAK board committee
A B S T R A C T S
Determination of GABAergic functioning in prefrontal cortex of early postnatal PCP rats
Hansen,C.K.1,2, Broberg,B.V.1,3, Kristiansen,U1 and Dalby,N.O.2
1.
2.
Department of Pharmacology and Pharmacotherapy, University of Copenhagen, Universitetsparken 2, 2100 Copenhagen.
Department of In Vivo Neuropharmacology, H. Lundbeck A/S, Otiiliavej 7-9, 2500 Valby.
3.
Center for Neuropsychiatric Schizophrenia Research, Copenhagen University Hospital, Psychiatric Center Glostrup.
Rats exposed to early postnatal treatment with the NMDA antagonist, phencyclidine (PCP), have been proposed as a
neurodevelopmental model of the deficits in executive function observed in schizophrenic subjects. Executive function
can be assessed by the attentional set-shifting task and after reaching adulthood the rats show selective impairment
in their extra dimensional shift performance. Among the strongest anatomical findings in adult rat is a reduction of parvalbumin (PA) in prefrontal cortex (PFC) suggesting a decline in the number of PA-positive interneurons. The aim of
the present project is to link these anatomical and behavioural findings. We hypothesize that PCP through a decrease
in PA-positive interneurons leads to a dysfunctional cortical network affecting the ability to perform executive function.
Functional characterisation of the inhibitory input to pyramidal neurons in PFC has been performed in adult Lister
Hooded rats (postnatal day 56-95) after early postnatal treatment with PCP (20 mg/kg, s.c.) or vehicle on postnatal day
7, 9, and 11. Coronal prefrontal cortical slices were superfused with ACSF containing 3 mM kynurenic acid. Whole-cell
voltage-clamp recordings were performed from pyramidal neurons located in layer II or V of medial PFC. Evoked inhibitory postsynaptic currents (IPSC) were recorded following a repeated paired pulse protocol with the stimulation electrode inserted in layer I of PFC. Inter-stimuli intervals were 100, 500 and 1000 ms. After application of 1 uM tetrodotoxin
(TTX) miniature IPSCs were measured for 8 min in the same cell.
A decrease in the frequency of miniature IPSC in pyramidal neurons of layer II of early postnatal PCP treated animals
(n=17) in relation to vehicle (n=13) were observed. Amplitude, rise time and decay time for the average event were
similar between the two groups. No difference in miniature IPSCs in layer V was evident. Paired pulse facilitation was
observed with 100 ms inter-stimulus interval (peak ratio~1.4) and paired pulse inhibition (peak ratio~0.9) with the 500
and 1000 ms inter-stimulus interval. There was no difference in the response between PCP and vehicle animals. The reduction in miniature IPSC frequency can indicate a decreased number of inhibitory synapses which correlates well with
a decreased number of PA-positive interneurons. It can also point towards a reduced presynaptic release probability
which indicate a functional deficit of existing interneurons. Both scenarios will have consequences for the functioning of
cortical networks which could lead to the observed behavioural changes.
A B S T R A C T S
Stereological quantification of parvalbumin positive interneurons in pre frontal cortex of
two animal models of schizophrenia
Riise J.L. 1, Kaalund S.1,3, Fabricius K. 2, Broberg B.V. 2,4, Owczarek S.1,3, Plath N. 2, Pakkenberg B1.
1.
Research Laboratory for Stereology and Neuroscience, Bispebjerg University Hospital, Bispebjerg Bakke 23, 2400 Copenhagen, NV Den
mark
2.
H. Lundbeck A/S. In vivo Neuropharmacology -. Ottilavej 7-9, 2500 Valby –DK.
3.
Protein Laboratory, Institute of Neuroscience and Pharmacology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark
4.
Center for Neuropsychiatric Schizophrenia Research, Faculty of health sciences, Copenhagen University Hospital, Psychiatric Center Glostrup, Denmark.
Schizophrenia is a complex and severe psychiatric disorder characterized by positive, negative and cognitive symptoms. These symptoms have been the basis in setting up animal models that mimic schizophrenia. However, as no
single animal model can represent the broad range of schizophrenic symptoms, several models have been developed.
Studies indicate a dysfunction of many neurotransmitters in the prefrontal regions of schizophrenia including glutamate
and GABA. Considerable attention has been drawn to the interaction of GABAergic interneurons and pyramidal cells
responsible for the synchronized gamma oscillations, which regulate working memory and information transmission
between cortical areas. In particular alterations related to fast spiking GABAergic interneurons containing the calcium
binding protein parvalbumin (PV) have been suggested as a core feature of the disorder that may account for a disruption of gamma oscillations and cognitive deficit found in schizophrenic patients.
In this study the optical fractionator design has been applied for stereological quantification of the PV positive cells as
well as the total number of neurons in the prefrontal cortex of two rat models of schizophrenia. These models include
the social isolated (SI)- and the neonatal phencyclidine (neoPCP) model that through environmental disturbances and
chemically induced alterations mimic the cognitive deficit associated with schizophrenia. Our preliminary results indicate
a tendency for a difference in the quantity of PV positive cells between control- and neoPCP-rats, which may suggest
PCP induced changes.
The current study validates the social isolation-and neoPCP model as models of schizophrenia and will provide useful
data for planning future cognitive translational studies.
A B S T R A C T S
Immunohistochemical investigations of parvalbumin, GAD65 and GAD67 protein levels in
a sub-chronic PCP model
Mark Holm Christensen1, Flemming F. Johansen2 and Uffe Kristiansen1
1.
2.
Faculty of pharmaceutical science, University of Copenhagen
Molecular Pathology Section, Department of Biomedical Sciences, BRIC, University of Copenhagen
Background and Purpose: Schizophrenia is a psychiatric disease affecting approximately 0.5 % of the worlds population. It consists of positive, negative and cognitive symptoms. The psychomimetic drug phencyclidine (PCP) can mimic
the symptoms of schizophrenia and sub-chronic PCP administrations to rats have shown to produce lasting cognitive deficit. In schizophrenic patients a decrease of parvalbumin (PV) expression in the dorsolateral prefrontal cortex
and hippocampus has been observed and associated with cognitive deficits. PV is a calcium binding protein found in
GABAergic fast spiking interneurons. There is also evidence of a decrease in GAD67 levels in the prefrontal cortex in
schizophrenics. Immunohistochemical investigations of PV and GAD levels were performed to assess the effects of
the sub-chronic PCP model.
Methods: 19 Female Lister Hooded rats were subjected to a sub-chronic administration of saline vehicle or PCP (2
mg/kg i.p. twice daily for seven days). Following a seven day wash out period brain sections were analyzed using immunohistochemistry. Stainings were made against PV and GAD65 in both the frontal cortex and the hippocampus and
stainings for GAD 67 were made in the hippocampus.
Results and Discussion: The density of PV immunoreactive (IR) interneurons were significantly increased in the
Cg1 (cingulate cortex area 1), Cg2 (cingulate cortex area 2), M1 (motor cortex area 1) and M2 (motor cortex area
2) of PCP treated rats compared to control rats. GAD65 (glutamic acid decarboxylase) IR neurons were significantly
increased in the Cg1, Cg2 and M1 in PCP treated rats. In the hippocampus an overall significant increase in PV IR
interneurons was also observed in the PCP treated group. Furthermore the colocalization of PV and GAD65 was also
significantly increased in the hippocampus of PCP treated rats. The results obtained in this study differ from the post
mortem findings in schizophrenic patients and behavioral changes seen in the sub-chronic PCP model one week post
PCP injections will not be the result of decreased PV interneurons.
A B S T R A C T S
Social Isolation of rats as a possible animal model of some traits of schizophrenia
Katrine Fabricius1, Lone Helboe2, Florence Sotty1, Anders Fink-Jensen, Gitta Wörtwein3, Bente Pakkenberg4 and B. Steiniger-Brach 1.
1.
2.
3.
H. Lundbeck A/S, Otiliavej 7-9, 2500 Valby, DK, Discovery Pharmacology Research and Mental Health Services Copenhagen, the Capi
tal Region of Denmark, University of Copenhagen.
H. Lundbeck A/S, Ottiliavej 7-9, 2500 Valby, DK, Discovery Biology Research.
Laboratory of Neuropsychiatry, Psychiatric Centre Rigshospitalet, University of Copenhagen, 2100 DK
4.
Research Laboratory for Stereology and Neuroscience, Bispebjerg University Hospital
Validation of animal models of psychiatric disorders is challenging due to the human nature of these disorders. The
isolation rearing model is based on the neurodevelopmental hypothesis of schizophrenia and has been used by others
as a model to mimic some of the deficits seen in schizophrenic patients. The aim of our studies was to validate the
isolation rearing model in Lister-hooded rats, using a combination of behavioural, stereological, neurochemical and
electrophysiological approaches. We isolated or group-housed (n = 5/cage) the animals on postnatal day 25 where
they remained isolated for a minimum of 8 -12 weeks.
With regards to mesolimbic dopamine neurotransmission, we report that isolated rats exhibited a spontaneous locomotor hyperactivity which could be reversed with antipsychotic. In vivo microdialysis studies showed unchanged
basal dopamine levels in the nucleaus accumbens (NAcc) but reduced in the prefrontal cortex (PFC) of isolated rats
versus group-housed rats. Further, isolated animals had an increased dopamine efflux in both the NAcc and the PFC
to an amphetamine challenge. We applied systematic uniformly random sampling (SURS) combined with the Cavalieri
principle to estimate the volume of different brain structures in the isolated and group-housed animals. We found that
isolated males had significantly smaller brain volumes and enlarged ventricles compared to group-housed controls,
and a reduced total hippocampal volume.
Taken together, the behavioural, electrophysiological and stereological findings are indicative of an overactive mesocorticolimbic system, show that the isolation rearing in Lister Hooded rats, at least to some extent, resembles schizophrenia -like abnormalities.
Type I interferons signaling in axonal injury-induced glial response
Khorooshi R and Owens T.
Institute of Molecular Medicine, University of Southern Denmark, Odense
Upon damage to the CNS, glial cells become activated. Such innate immune response is critical for the induction of
cytokines and chemokines that direct leukocytes entry to the injury site. These responses may contribute to repair processes in the damaged CNS, but may also exacerbate neurodegeneration. To better understand these processes, it
is important to study the mechanism of signaling in glial responses to injury. The glial response may involve cytokines,
such as type I interferons (IFNα/β), which are known to regulate innate immune responses against viral infections.
IFNα/β signal through a receptor (IFNAR), which involves activation of STAT1/2 and interferon regulatory factor-9
(IRF9), leading to activation of interferon stimulated genes including IRF7. The induction of IFNα/β is enhanced by
IRF7. We have examined the involvement of IFNα/β signaling in hippocampus after axonal transection. Axonal injury
induced up-regulation of IFNα/β receptor associated signaling in hippocampus. Double immunofluorescent co-localization studies showed that IRF7 was induced in Mac-1/CD11b positive macrophages/microglia in the denervated
molecular layer of dentate gyrus. In addition, IRF7 mRNA was detected in FACS-sorted microglia from lesion-reactive
hippocampi. The induction of IRF7 mRNAs was IFNAR-dependent. Furthermore, lack of IFNα/β signalling resulted in
increased leukocyte infiltration into the lesion-reactive hippocampus. Unlike WT mice, axonal lesion did not induce an
increase in CXCL10, but it induced an increase in matrix metalloproteinase 9 gene expression in IFNAR-KO mice. Our
findings point to a role for type I IFN signalling in regulation of innate immune response to sterile injury.
A B S T R A C T S
The role of neuronal nuclear factor-kappa B after moderate spinal cord injury.
Ellman D1, Bracchi-Ricard V2, Bethea JR2 and Lambertsen KL1
1.
2.
Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, 5000 Odense C, Denmark and
The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL, 33136, USA.
The goal of our study is to investigate the contribution of neuronal nuclear factor-kappa B (NF-κB) signaling to both the
secondary injury process and the functional recovery following spinal cord injury (SCI). NF-kB is a family of transcription factors kept inactive in the cytoplasm as dimers bound to an inhibitor of kB (IkB) molecule. When NF-kB signaling
pathway is activated (i.e. by cytokines), phosphorylation of the IkB Kinases (IKK) complex (consisting of IKK1, IKK2 and
NEMO) occurs, which in turns phosphorylates IkB targeting it for ubiquitination and degradation via the proteasome.
This leads to the release and translocation of the NF-kB dimer to the nucleus where it can activate gene transcription.
NF-κB signaling is critical for the regulation of immune responses, inflammatory reactions and apoptosis and is also
involved in some of the pathophysiological changes observed following brain injury. Selective inhibition of NF-κB in
neurons, but not astrocytes, leads to reduced infarct volume in a mouse model of focal cerebral ischemia. This indicates
that neuronal NF-κB activation can be damaging. We have shown that selective inhibition of astroglial NF-kB activation
leads to reduced inflammation, smaller lesion volume and improved functional recovery 8 weeks following SCI. However
the role of NF-kB signaling in neurons following SCI has not yet been studied.
Our hypothesis is that inhibition of neuronal NF-κB signaling will lead to a reduced neuroinflammatory response, which
will result in reduced secondary injury and increased functional recovery after SCI. To test this hypothesis we will delete
the IKK2 subunit of the IKK complex specifically in neurons by crossing IKK2FL/FL mice with mice expressing the Cre
recombinase enzyme under the control of the synapsin promoter. While it isn’t possible to intervene against the primary,
mechanically conditioned tissue damage after SCI, animal studies show that it is possible to obtain functional recovery
after moderate SCI by intervention against the secondary injury mechanism, which is crucial for the long-term survival
of the nerve cells some distance away from the epicenter.
A B S T R A C T S
The influence of parenchymal damage on cortical spreading depression in patients with
severe brain injury.
Nina Eriksen1, Egill Rostrup2, Kjeld Benda1, Martin J. Lauritzen3, Martin E. Fabricius3, Vibeke A. Larsen4, Jed A. Hartings5, Jens P. Dreier6, Anthony J.
Strong7, and Bente Pakkenberg1
1.
2.
3.
4.
5.
6.
Research Laboratory for Stereology and Neuroscience, Copenhagen University Hospital Bispebjerg, Denmark
Department of Physiology and Nuclear Medicine, Copenhagen University Hospital Glostrup, Denmark
Department of Neurophysiology, Copenhagen University Hospital Glostrup, Denmark
Department of Radiology, Copenhagen University Hospital Glostrup, Denmark
Department of Neurosurgery, University of Cincinnati, OH, USA
Charité University Medicine Berlin, Germany
7.
King’s College Hospital, London, United Kingdom
Severe brain damage is often followed by serious complications for the patient. In traumatic and ischaemic brain injury, a phase of delayed deterioration often associated with severe and refractory brain swelling sometimes develops
between 2 and 5 days after the initial ictus, and is associated with poor or fatal outcome. It has become evident mainly
from experimental work that permanent and transient depolarisations (peri-infarct depolarisations and cortical spreading depression) of grey matter of the central nervous system may be major constituents of progressive deterioration in
border zones of traumatic or ischaemic foci. Quantitative measurements, such as regional volumes and surface areas
under various conditions are essential for understanding functional changes in the brain, and assessing prognosis. As a
part of a larger study, COSBID – CoOperative Study on Brain Injury Depolarizations on traumatic and ischaemic injury
to the human brain, we focus on volume estimation of traumatic regions as a prognostic tool.
The affected brain tissue in patients suffering from subarachnoid haemorrhage (SAH) or traumatic brain injury (TBI)
is very inhomogeneous; hence traditional methods used in imaging are not useful. Therefore, volumetry of the injured
regions during acute, subacute and follow up stages was obtained in-vivo from CT and MR images by Cavalieri method,
and correlated to the events of depolarizations.
There was a tendency for slightly larger damaged area in the sub-acute phase, which supports the theory of the progressive deterioration in the traumatic areas. The accuracy of the stereological methods shows that they are highly reliable
tools for quantification, and more precise information from head trauma patients can be obtained from stereological
measurements and used for evaluation of treatment and prognostic value.
Coupling migraine and inflammation? A study in rat trigeminal ganglia
Kim Anker Kristiansen and Lars Edvinsson
Glostrup Research Institute, Glostrup Hospital, Ndr Ringvej 69, DK-2600 Glostrup, Denmark
There is a growing body of evidence that migraine and inflammation are linked together, often termed as neurogenic
inflammation. Activation of the trigeminovascular system has been shown to lead to the release of vasoactive neuropeptides such as calcitonin gene-related peptide (CGRP) and substance P (SP), while acute migraine attacks only results
in CGRP release
Since previous studies mainly involved either culture of TG neurons or SGC our aim of this study was to study the effects of culture of intact trigeminal ganglia sections in relation to regulation of pro- and anti-inflammatory cytokines on
mRNA level. Furthermore, since some SGC and neurons have CGRP receptors the influence of CGRP and its inhibitor
CGRP7-38, and several MAPK pathway blockers, have been assessed. Finally, activated MAP kinases, cytokines and
cytokine receptors were studied at protein level using immunohistochemistry.
A B S T R A C T S
Do neuroglobin protect neurons during focal brain ischemia?
Zindy Raida
Since the mid 80´s the endovascular filament middle cerebral artery occlusion (MCAo) approach has been widely used
in studies of transient brain ischemia in rats. Because of a very small vessel size the MCAo approach has not been
frequently used in mice, but due to an increased usage of genetic modified mice in stroke research there is now a proliferated demand to adopt and refine the MCAo approach for mice. I will describe a simple method for intratracheal intubation in mice for mechanical ventilation without use of injection anesthesia for immobilization during intubation. Describe
the MCAo operational approach in mice and briefly address the critical factors causing outcome variation. These critical
factors include anesthetics, maintenance of animal physiology and model specific factors that affect success rate and
variation.
Noradrenaline release: a potential antidepressant mechanism of brain stimulation therapies
Anne M Landau, Suzan Dyve, Aage Kristian Olsen Alstrup, Arne Møller, Mette Simonsen, Steen Jakobsen, Poul Videbech, Gregers Wegener, Albert
Gjedde, and Doris J Doudet
Brain stimulation therapies are currently being used to treat depression in drug-resistant patients. Vagal nerve stimulation (VNS) involves direct stimulation of the vagus nerve, and although it first emerged as a treatment for intractable
epilepsy, it is currently recognized as a potentially useful mood elevation treatment. Electroconvulsive therapy (ECT),
a direct form of electrical brain stimulation, is the most effective antidepressant therapy known. We tested the hypothesis that the beneficial effects of brain stimulation therapies are mediated, at least in part, through augmentation of the
inhibitory effects of cortical monoaminergic neurotransmission, specifically noradrenaline. We established two novel
models of brain stimulation in Göttingen minipigs, based on human treatment protocols. Using the positron emission
tomography (PET) ligand [11C]-yohimbine, an alpha2 adrenoreceptor antagonist, we determined the changes in adrenergic neurotransmission associated with VNS and ECT brain stimulation. In anesthetized minipigs, PET scans were first
acquired 4-6 weeks after the implant of a VNS stimulator before (OFF condition, baseline), within 30 minutes after the
initiation of stimulation at a current intensity of 1mA (ON condition, acute), and after 3 months of ramped up stimulation
to 2mA (ON condition, chronic). Similarly, we acquired baseline scans in naïve animals and 24-48 hours after the end of
a clinical course of ECT (10 ECT sessions in anesthetized animals over a 3 week period). Preliminary data demonstrate
decreased volumes of distribution of [11C]yohimbine to alpha2 adrenoceptors compared with the baseline scans in both
stimulation models. Kinetic analysis using the Logan multilinear graphical analysis method in the first three pigs of the
VNS study, resulted in a decrease in volume of distribution in the frontal cortex (7% acute, 22% chronic), temporal cortex
(12% acute, 22% chronic) and striatum (15% acute, 29% chronic). Data from the first three pigs in the ECT study show a
decrease in volume of distribution in the frontal cortex (29%), temporal cortex (25%) and striatum (31%) in ECT treated
pigs, 24-48 hours after the final ECT treatment. Taken together with published evidence in rodents, these findings suggest that acute and chronic brain stimulation may affect the cortico-striatal noradrenaline system and suggest a possible
general mechanism of action for brain stimulation therapies such as ECT and VNS.
A B S T R A C T S
Viral vector-induced overexpression of neuropeptide Y receptors in treatment of seizures
Casper R Gøtzsche, Mikkel V Olesen, Søren H Christiansen, David PD Woldbye
Laboratory of Neuropsychiatry, Department of Neuroscience and Pharmacology and Psychiatric Centre Copenhagen – University of Copenhagen,
Denmark
Gene therapy with adeno-associated viral (AAV) vectors inducing overexpression of neuropeptide Y (NPY) in the hippocampus leads to seizure-suppressant effects. This approach might be used in future treatment of temporal lobe
epilepsy. NPY Y2 and Y5 receptors play a major role at mediating antiepileptic effects of NPY in the hippocampus and
overexpression of Y2 or Y5 receptors could be a more specific alternative to overexpressing NPY alone. Using an AAV
vector, long-term overexpression of functional Y2 and Y5 receptors was induced in the hippocampus of adult rats, and
the effects were studied in temporal lobe epilepsy models: rapid electrical kindling and/or kainate-induced seizures.
Moreover, it was also tested whether combined AAV-induced overexpression of an NPY receptor and NPY could result
in a better antiepileptic therapeutic response than with the single gene approach. Y2 overexpression resulted in seizuresuppressant effects in both models. Inhibitory effects were also found after NPY-AAV treatment, consistent with previous
studies. However, combined overexpression of Y2 and NPY enhanced the seizure-suppressant effect as compared to
that of single transgenes with regard to several seizure parameters. In contrast, transgene Y5 overexpression alone
exerted no effect on seizures while combined overexpression of Y5 and NPY caused antiepileptic effect. These results
show for the first time that Y2 receptor-based gene therapy could be a novel treatment strategy in epilepsy and that
combining Y2 or Y5 with NPY overexpression could improve the seizure-suppressant response.
Comparison study of the human brain using histology, stereology and MRI
Anna S. Karlsen1, Tim B. Dyrby2 & Bente Pakkenberg1
1.
2.
Research Laboratory for Stereology and Neuroscience, Copenhagen University Hospital, Bispebjerg, Denmark
Danish Research Centre for Magnetic Resonance, Copenhagen University Hospital, Hvidovre, Denmark
Mental retardation is a consistent phenotype in many developmental disorders. Down syndrome (DS) or trisomy 21
involves mental impairment, delayed growth, accelerated aging and dementia and is a model for investigating the physical consequences of genetic anomalies. Volume and cell density were estimated in the neocortex and its four major
sub-divisions using the stereological tools - the Cavalieri estimator of volume and the optical disector in adult female DS
subjects (mean age 69.3 years, N = 5) and compare with female controls (mean age 70.0 years, N = 6).
A total number of 11.1 × 109 neocortical neurons (coefficient of error = CE = 0.04, range 9.55 - 12.0 × 109) was estimated and compared to controls having 17.8 × 109 neurons (CE = 0.04, range 14.4 - 21.1 × 109); showing a global
difference of almost 40 %, which was highly statistically significant (2p = <0.001). This lowered number of neocortical
cells was the result of a diminished neocortical volume with no changes in cell densities.
Despite of a general agreement on total brain volume in normal brains when comparing estimates from stereology
and MRI, an overestimation of cortical volume by 15 - 20 % was observed in structural MRI (submitted work, personal
communication). Motivated by this observation, we investigated differences in image information in structural MRI and
physical sectioning. Eight of the formalin-fixated brains (four controls and four DS) were scanned before sectioning in a
clinical 3T Siemens TRIO MR scanner using a 3D T1-weighted sequence. These scans revealed structural aberrations
such as subcortical hypo-intense areas as well as cortical atrophy, which were structurally analyzed. The outcome of the
volumetric comparison study is presented and will indicate the sensitivity of 3T MRI to detect abnormalities.
A B S T R A C T S
Virtual slides: Development of automatic neuro-analysis tools and verifying them using stereology
Allan Rasmusson1, Paulette Herlin2, Benoir Plancoulaine2, Nicolas Petit2, and Jens Randel Nyengaard1
1.
Stereology and EM Laboratory, Aarhus Hospital NBG, Aarhus
2.
GRECAN, Centre Francois Baclesse, Caen, France
Abstract: The advent of scanners and microscopes able to virtualize entire slides of tissue samples at high magni_cation
brings forth the possibility to develop software that automatically performs analyses and classi_cation of tissue. This
talk will enlighten the steps needed in development of tools for automatic analysis of blood vessels in brain tumors. The
steps are: 1.Handling large images: The recorded virtual slides often reach sizes in excess of the memory available to
a workstation. To overcome this they can be split into smaller manageable parts or be analyzed at lower magni_cation.
2. Developing tools for analysis: Using digital image processing programs for identifying neurons, blood vessels etc. on
the slides can be developed. 3. Veri_cation of the developed tools: It is of utmost importance to verify that the programs
processing the virtual images yield correct results. Essentially, an expert must evaluate the performance at
every part of the virtual slide. This is a time very consuming task, but fortunately existing stereological tools can decrease the amount of work.
The effect of Interferon regulatory factor 7 on the development of experimental autoimmune encephalomyelitis in mice
Mohammad Salem, Reza Khorooshi, Trevor Owens
Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense
Interferon regulatory factor 7 (IRF7) is critical for the induction and positive feedback regulation of type I interferon
(IFNα/β) responses. IFNβ is used as a therapeutic for multiple sclerosis, but little is known about the basic mechanism
behind the action of IFNα/β and IRF7 in CNS physiology or pathology. The aim of the present study was to examine the
impact of IRF7 on the development of experimental autoimmune encephalomyelitis (EAE) in mice. IRF7 gene expression increased in the CNS of C57BL/6 (wild-type) mice with EAE. IRF7-deficient mice developed more severe EAE.
FACS analysis showed that both WT and IRF7-KO mice had increased leukocyte infiltration into the CNS. However,
IRF7-deficient mice had significantly higher number of infiltrating macrophages and T cells in the CNS compared with
WT. This was further confirmed by immunohistochemistry which also showed altered distribution of infiltrates within the
spinal cord. Analysis of chemokine gene expression by quantitative real-time PCR showed that both WT and IRF7deficient mice had increased CCL2 and CXCL10 gene expression in the CNS. However, the level of these chemokines
was higher in IRF7-deficient mice compared to WT mice. Together, our findings suggest the involvement of IRF7 in
autoimmune inflammation in the CNS.
A B S T R A C T S
ThTranscriptional regulation of myelin formation in adult CNS
Christina Fenger1, Mads Thomassen2, Torben Kruse2 og Bente Finsen1.
1.
2.
Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense,
Human Microarray Center, Department of Clinical Genetics, Odense University Hospital.
A number of neurological diseases involve acquired or congenital defects in CNS myelin. The formation of functional
CNS myelin depends on successful differentiation of oligodendrocyte progenitor cells (OPCs) into myelinating oligodendrocytes (OLs) which is under strict genetic regulation. In vitro studies have shown that the Nkx2.2 transcription factor
activates the expression of proteolipid protein (PLP), but represses the expression myelin basic protein (MBP) which
both are myelin structural proteins essential to the formation of functional myelin. Since Nkx2.2 is up-regulated during
remyelination, it is crucial to determine, if Nkx2.2 inhibits or promotes remye-lination in vivo, and to know the Nkx2.2
target genes implicated in OPC differentiation and myelination/remyelination.
We initially examined the effect of Nkx2.2 remyelination in vivo. Loss of myelin and OLs was induced by treatment of
young adult Nkx2.2+/- mice and littermate wild type (Wt) mice with a diet containing 0.4% cuprizone for 6 weeks. Remyelination was spontaneously induced by 2 additional weeks of recovery on standard diet. Nkx2.2-/- mice die early
postnatally and could therefore not be used for the study. We found that Nkx2.2+/- mice showed a delayed, possibly
generally reduced, remyelination response which was manifested by fewer NogoA+ OLs in the corpus callosum of
Nkx2.2+/- mice compared to Wt mice after 6 weeks of cuprizone treatment, and lower LFB+ myelin score after additional
2 weeks of recovery on standard diet.
We next investigated the potential Nkx2.2 target genes by comparison of the mRNA profile in brainstem in newborn
Nkx2.2-/- and Wt mice using microarray technology followed by QPCR based validation of results. We have identified
17 putative Nkx2.2 target genes, most of which are OL specific genes, including the two previously discovered Nkx2.2
target genes, PLP and MBP. The results indicate that Nkx2.2 promotes the expression of all the putative Nkx2.2 target
genes, since these are all down-regulated in Nkx2.2-/- mice compared to Wt mice. Our next task is to identify the primary Nkx2.2 target genes by ChIP-on-chip technique and to confirm that the primary target genes co-localize to Nkx2.2
expressing OL lineage cells both during myelination of mouse brainstem and during remyelination of the adult mouse
brain.
So far, we can conclude that Nkx2.2 promotes the formation of OLs in both the developing and adult CNS, and we have
identified 17 potential Nkx2.2 target genes. We hope that the project will contribute to greater molecular understanding
of the myelination/remyelination processes with the prospect to improve the capacity of the CNS to remyelinate.
Interferon-β in CNS homeostasis and neurodegeneration: A matter of impaired autophagy
Christian Berg
Interferon-β (IFNβ) is a pleiotrophic cytokine secreted by most cells in response to recognition of foreign antigens.
Recombinant IFNβ is at present an important treatment modality for multiple sclerosis (MS), yet only a subset of patients
respond to the treatment. To develop more efficient drugs it will be necessary to understand the mechanisms behind the
clinical effect of IFNβ.
We have promising data suggesting that in addition to immune modulation, IFNβ also acts as a neuroprotectant. Accordingly, the lack of IFNβ results in spontaneous neurodegeneration with pathology similar to neurodegenerative diseases
with Lewy Body Dementia such as Parkinson’s disease (PD) and Alzheimer’s disease (AD).
Here I will present my working model where IFNβ mediated induction of Cyclin Kinase Inhibitors (CKI), such as p16, p21, p27
promotes survival in neurons through inhibition of the E2F pathway. CKIs serve to restrict cell proliferation and are crucial for
keeping neurons in a post-mitotic state. In vivo and in vitro experiments have established that deregulation of the E2F pathway
lead to neuronal cell cycle re-entry and subsequently apoptosis. Interestingly, several pharmacological cell cycle blockers are
neuroprotective mice challenged with acute injury.
In order to address this hypothesis, I will use cultured mouse cerebellar granule cells to answer whether IFNβ promote neurons
survival through induction of CKI’s.
A B S T R A C T S
The role of Interferon-β in CNS homeostasis and neurodegeneration:
A matter of impaired autophagy?
Martin Kuss, Malene Ambjørn, Shohreh Issazadeh-Navikas
BRIC, Copenhagen Biocenter, Copenhagen, Denmark
Interferon β (IFN-β) is a pleiotropic cytokine which is secreted by many cells during pathogeninfection as well as in
response to inflammatory stimuli propagating anti-inflammatory properties. Mice lacking expression of IFN-β are more
susceptible to develop a severe experimental autoimmune encephalomyelitis (EAE), a commonly used mouse model of
multiple sclerosis (MS). Moreover, depletion of IFN-β leads to spontaneous signs of neuronal degeneration with striking
similarity to the pathology of Lewy body dementia, Alzheimer’s and Parkinson’s disease. This indicates that endogenous
IFN-β participates in the maintenance of neuronal homeostasis.
Gene expression profiling by micro array analysis of cultured cerebellar granule neurons (CGNs) from IFN-β knock out
(ko) mice proposed a possible involvement of autophagy in IFN-β dependent homeostasis. Autophagy is a lysosomal
bulk degradation pathway originally described as a starvation response. However, recent evidence emphasizes the protective role of autophagy in neuronal homeostasis, probably due to its constitutive clearance of cytosolic proteins which
otherwise would accumulate and/or aggregate, ultimately causing neurodegeneration.
RT-qPCR analysis of neuronal cultures was performed on several genes involved in autophagy and its regulation. Irgm1
and Igtp were found to be highly inducible by IFN-β and decreased in IFN-β ko neurons compared to wildtype (wt).
An enzymatic assay addressing the function of cathepsins, which are lysosomal cysteine proteases and important in
cleavage of autolysosomal contents, showed that IFN-β ko brain tissue display slightly reduced cathepsin B/L activity
compared to wt. Immunohistochemical analysis of IFN-β ko compared to wt brains was pointing towards accumulation
of the autophagy marker p62, yet not significantly.
Together with other data from out lab, this is supporting the notion that IFN-β is playing an important role in CNS homeostasis by regulation of autophagy.
A B S T R A C T S
CD1-deficient mice develop spontaneous gait ataxia and depressive symptoms
Jeanette Svensson, PhD student
Supervisor Professor Shohreh Issazadeh-Navikas
Neuroinflammation Unit, Biotech Research and Innovation Centre (BRIC), Biocentre of Copenhagen, University of Copenhagen, Denmark
The central nervous system (CNS) has been considered to be an immune-privileged site, preventing immune cell
access in order to protect the tissue from irreversible damage. However, almost all aspects of CNS tissue damage,
including autoimmunity, such as classical inflammatory CNS diseases e.g. Multiple sclerosis (MS) as well as classical
neurodegenerative diseases e.g. Alzheimer’s (AD) and Parkinson’s disease (PD) and their animal models, involves a
significant cellular inflammatory component since activated lymphocytes, which have escaped immune tolerance, have
the ability to penetrate the blood-brain barrier.
Cluster of differentiation 1 (CD1) mainly presents glycolipidic antigens. In mice, the CD1 molecule is expressed on
antigen-presenting cells, but its expression has also been observed on T lymphocytes, microglia, astrocytes and possibly even on neurons. CD1 have been shown to be of importance in inducing tolerance and controlling autoimmunity.
Results from the group has shown regulatory functions of CD1-restricted NKT (natural killer T cells) cells demonstrating
that EAE (experimental autoimmune encephalomyelitis), an animal model of MS, mice deficient in CD1 were associated
with increased peripheral encephalitogenic T cell cytokine production and diminished levels of the anti-inflammatory
cytokine TGF-β1 in the CNS. Novel reports from the group have also demonstrated spontaneous behaviour alterations
in CD1-deficient mice such as hypoalgesia, ataxia and a depression-like behaviour, symptoms that are commonly seen
in CNS inflammatory and neurodegenerative diseases such as MS, AD, PD and in their animal models.
In addition, the altered behaviour was not correlated an imbalance in peripheral immune cells, indicating that the peripheral immune system presumably play a negligible role in the phenotype. The phenotype could however be correlated
with an CNS increase in quantitative mRNA expression of TNF and TGF-β1 and decreased levels of neurofilament,
ubiquitin, MHC class I and II, and GABA (gamma-aminobutyric acid) A receptors.
The aim of my project is to investigate why the absence of CD1 causes more severe EAE and induce spontaneous
behaviour abnormalities by investigating the role of CD1 immune-regulatory role in spontaneous and induced (EAE)
CNS inflammation and the role of CD1 in neuronal homeostasis.
Pigs in PET: Differing effects of propofol versus isoflurane on dopamine 1 receptor binding
in Göttingen minipigs.
By Aage Kristian Olsen Alstrup & Annie M. Landau. PET-centret i Århus
The dopamine 1 (D1) receptor has in recent years become a site of interest for its involvement in a number of psychiatric conditions. It’s characteristics and modulation are extensively studied in experimental models and pharmacological
challenges using the in vivo capabilities of PET imaging. The role of various anesthetics on binding of PET ligands to
the dopaminergic D2/3 receptors has been carefully examined but little data exist on the effect of anesthesia on binding
parameters of D1 receptors. In this study, we evaluated the role of two routinely used anesthetics on the binding of a
common tracer of the D1 receptor, SCH23390, using Göttingen minipigs, which are attracting increasing interest for in
vivo imaging due to their behavioral characteristics and large brain size. Minipigs were anesthetized with either isoflurane (n=5) or propofol (n=3) during PET studies with the D1 receptor tracer, [11C]SCH23390. Using the Logan reference
tissue model, the SCH23390 binding potential in the isoflurane-anesthetized group was significantly higher than in the
propofol-anesthetized group in the striatum (59 %), putamen (55 %), caudate (63 %), thalamus (39 %), temporal cortex
(53 %), frontal cortex (60 %), and globus pallidus (51 %). The magnitude of the differences in these data enlightens the
importance of the effects of anesthesia during animal experimentation and on the possible interpretation of the data.
It remains to be determined how these responses of the dopamine D1 receptors to various anesthetics may affect the
measurements of D1 binding during pharmacological or therapeutic challenges.
A B S T R A C T S
The Story of Music and Dopamine – A pleasurable surprise
Line Gebauer
Music listening is extremely pleasurable and studies show that music listening activates structures associated with the
mesolimbic dopaminergic reward system. However, how music is capable of inducing pleasure and recruiting the brain’s
reward system is not very well described in the literature. In this presentation, I propose anticipation as a fundamental
mechanism behind musical pleasure, and dopamine as an important mediator between music and pleasure. Dopamine
is critically involved in anticipation, especially with regards to reward anticipation and registration of prediction errors,
which occur when the anticipations are not fully met. Dopamine is likewise suggested to be involved in pleasure, but here
I suggest that dopamine merely mediates musical pleasures through anticipation and hence do not cause the pleasure
per se. Consequently, the presented account of anticipation in music integrates both psychological and neurobiological
mechanisms underlying musical pleasure. A better understanding of the anticipatory interplay between musical structures and expectations in the listener, may bring us closer to an understanding of both how and why music is capable
of inducing strong emotions and pleasure in the listener, and thus why music is an essential part of most people’s lives.
Dopamine Transporter Knock-In Mice with a Disrupted PDZ-Protein Binding Motif Display
Hyperactivity and Impaired Amphetamine Response
G. Sørensen1,2, M. Rickhag1, K. Nørgaard-Strandfelt1, B. Andresen1, E.M. Füchtbauer3, J. Gomeza4, G. Wörtwein2, D. Woldbye2 and U. Gether1
1.
University of Copenhagen, Department of Neuroscience and Pharmacology, Molecular Neuropharmacology Group, Copenhagen, Den
mark.
2.
University of Copenhagen, Department of Neuroscience and Pharmacology & Mental Health Centre Rigshospitalet, Laboratory of Neu
ropsychiatry, Copenhagen, Denmark.
3.
Aarhus University, Department of Molecular Biology, Aarhus, Denmark.
4.
University of Bonn, Institute for Pharmaceutical Biology, Bonn, Germany.
Dopamine has an important modulatory role in the central nervous system and an impairment of function is associated
with several neuropsychiatric diseases. Re-uptake from the extracellular compartment by the dopamine transporter
(DAT) is crucial for homeostatic transmitter levels. Recent data suggest that DAT is part of a highly complex multi-protein
network controlling critical aspects of DAT function including synaptic distribution, compartmentalization and surface
targeting. In an attempt to investigate the role of specific protein-protein interactions for DAT function, we generated
knock-in mice expressing a mutant DAT with perturbed ability to interact with PDZ-domain proteins.
In a general behavioural test, the SHIRPA primary screen, the DAT knock-in mice showed no differences as compared
to wild-type mice. However, the DAT knock-in mice displayed increased spontaneous locomotor activity and showed a
striking absence of locomotor response to amphetamine treatment (2 mg/kg i.p.). The lack of amphetamine response
was found not only after injections immediately prior to testing but also seen when the mice were injected with amphetamine after 2½ hours of habituation to the activity boxes.
Our results suggest that disruption of PDZ-protein interactions has profound influence on DAT function as evidenced by
hyperactive knock-in mice lacking behavioural response to amphetamine.
A B S T R A C T S
A prospective study of epileptogenesis, seizure semiology, therapeutic outcome, mortality
and risk factors in a population of dogs with newly diagnosed epilepsy
Nadia Fredsø Andersen1, Anne Sabers2, Arne Møller3, Mette Berendt1
1.
2.
3.
Department of Small Animal Clinical Sciences, Faculty of Life Sciences, University of Copenhagen
Epilepsy Clinic – The neuroscience Center, Rigshospitalet
PET-centre, Aarhus University Hospital and Center for Functionally Integrative Neuroscience, Aarhus University
In dogs, epilepsy is the most common neurological disorder with an estimated prevalence of around 1%. In dog breeds
with genetic epilepsy the prevalence is however much higher as e.g. in the Belgian Shepherd 9.5%. The severity of the
condition is reflected in the fact that seizure freedom is difficult to achieve medically, and in both humans and animals
there is an increased risk of premature death.
Although “old” drugs (introduced in human medicine in the early 1900s and in the mid 1800s), Phenobarbital and Potassium bromide – because of their long half-lives – are the only drugs suitable for treatment of dogs with epilepsy. It has
been advocated that treatment with Phenobarbital possess a higher risk for serious side-effects than Potassium bromide. A recent pilot study from Department of Small Animal Clinical Sciences at LIFE-KU has confirmed this suspicion.
Unfortunately many myths exist in veterinary medicine with respect to canine epilepsy, and therefore there is a great
need for controlled clinical studies that can document the true nature of the disease.
The aim of the current study is to investigate epileptogensis, seizure phenomenology, risk factors, and mortality as well
as efficacy and side-effects of antiepileptic treatment in a cohorte of drug naïve dogs with newly diagnosed epilepsy.
Dogs will be followed in a prospective longitudinal study for two years. 60 dogs will be included in the study.
The treatment study will be carried out as a prospective randomized double blinded treatment study where a subpopulation of drug naive dogs with newly diagnosed epilepsy will be randomized to treatment with either Phenobarbital
or Potassium bromide. Efficacy and side-effects of the two drugs will be studied and compared over a two year period.
A subgroup of the included dogs will undergo a MR- and PET scan to look for potential morphological and metabolic
brain changes compared to a group of dogs with chronic epilepsy.
Characteristics and phenomenology of epileptic partial seizures in dogs: similarities with human seizure semiology. Epilepsy Research, 2004;61:167173.
Berendt M, Gredal H, Ersbøll AK, Alving J. Premature death, risk factors and life patterns in dogs with epilepsy. J Vet Int Med 2007; 21:754-759.
Berendt M, Gulløv CH, Christensen SLK, Gudmundsdottir H, Gredal H, Fredholm M, Alban L. Prevalence and characteristics of epilepsy in the Belgian
shepherd variants Groenendael and Tervueren born in Denmark 1995-2004. Acta Vet Scand 2008; 50:51.
Berendt M, Gulløv CH, Fredholm M. Focal epilepsy in the Belgian shepherd: Evidence for Mendelian inheritance. J Small Anim Pract 2009;50:655661.
A B S T R A C T S
Role of M4 muscarinic receptors in psychostimulant-mediated effects
Ditte Nielsen1, Pia Weikop1, David P.D. Woldbye1, Jürgen Wess2, Gitta Wörtwein1 and Anders Fink-Jensen1
1.
2.
Laboratory of Neuropsychiatry, Department of Neuroscience and Pharmacology and Psychiatric Centre Copenhagen – University of Copenhagen, Denmark
Laboratory of Bioorganic Chemistry, National Institutes of Health, NIH-NIDDK, Bethesda, USA
Central actions of acetylcholine are mostly mediated by muscarinic (M1–M5) receptors and disturbances in the central
muscarinic cholinergic system have been implicated in several pathophysiological conditions, including drug addiction,
and schizophrenia, conditions that involve the dopaminergic system. Behavioral and neurochemical studies of M4 receptor knock out mice have shown that the M4 receptor subtype plays an important role in the regulation of dopamine
neurotransmission.
We wanted to further investigate the involvement of muscarinic M4 receptors in the behavioural and neurochemical effects of psychostimulants that release dopamine. To this end we investigated cocaine-induced hyperlocomotor activity
and cocaine-induced striatal dopamine release in fully back-crossed M4 receptor knockout mice. We also investigated
the effect of a potent and selective allosteric potentiator of M4 receptors (VU0152100) in these models in wild type mice.
M4 receptor deletion enhanced cocaine-induced locomotor activity and cocaine-induced dopamine release in the nucleus accumbens. In accordance with this observation, the selective allosteric potentiator of M4 receptors VU0152100
inhibited cocaine-induced hyperactivity and striatal dopamine release in wild type mice. The present data suggest the
M4 muscarinic receptor as a possible new target in the treatment of drug addiction and psychosis.
Microglial Response in a Mouse Model for Cerebral Amyloidosis
Rebecca Radde1,2
1.
Institute for Molecular Medicine, SDU, Odense,
2.
Hertie-Institute for Clinical Brain Research, Tuebingen, Germany
A hallmark lesion in Alzheimer’s disease pathology is the aggregation of Aβ peptides into extracellular amyloid deposits.
To model this cerebral amyloidosis we have generated a transgenic mouse model by the overexpression of mutated
amyloid precursor protein (APP) bearing the “Swedish” double mutation KM670/671NL and L166P mutated presenilin
1 (PS1) under control of the neuron-specific Thy1.2 promoter element. Resulting APPPS1 mice exhibit high fibrillogenic
Aβ42 levels, and dense congophilic amyloid assemblies from an age of six weeks on. Even newly emerging deposits are
associated with dystrophic neurites, reactive microglia and astrocytes. Amyloid burden progressively aggravates with
aging accompanied by exacerbated neuroinflammatory response.
Our work sought to investigate the microglial response after amyloid lesioning. We used stereological methods to
quantify the microglial cell number in APPPS1 mice. Microglial cells were triplicated in 8 months old amyloid depositing
mice compared to 1 month old pre-depositing animals. In vivo imaging techniques reveal a rapid movement of microglia
processes towards the amyloid insult, followed by the migration of the microglia cell body to the plaque surface where it
remains stable over weeks. The plaque volumen interrelates to the microglial cell size and reveals signs of phagocytotic
activity. Along this line, we found an incorporation of congophilic dye into microglial cells indicating signs of amyloid
uptake. Crossing APPPS1 on an CD11b-HSKtk background allows the selective ablation of microglia by ganciclovir application. Microglial depletion in APPPS1 mice for four weeks did not change amyloid plaque formation. Together, these
findings indicate the inefficient clearance of entire amyloid deposits through microglia in this model, although amyloid
incorporation can be observed on a cellular level.
A B S T R A C T S
Expression of IL-1β and TNF-α in Alzheimer’s pathology in mice before and after endotoxin
challenge
Ilkjær L1, Babcock A1, Skovgård M2, West M2, and Finsen B1.
1.
2.
Institute for Molecular Medicine, University of Southern Denmark, Odense.
Institute of Anatomy, University of Aarhus, Aarhus C.
Chronic activation of microglia, possibly by amyloid beta (Aβ) plaques, is significant in Alzheimer’s disease (AD), and
might play a central role in its pathogenesis. Activated microglia produce potentially neurotoxic substances including the
cytokines tumour necrosis factor α (TNF-α) and interleukin-1β (IL-1β). Here, we investigated the mRNA levels of TNF-α
and IL-1β in an APP/PS1 double transgenic (Tg) mouse model for AD in the age groups 3, 6, 9, 12, 15, 18, 21 and 24
months. Since inflammation is believed to play a damaging role in AD we also investigated the effect of 1 and 3 months
treatment with an endotoxin on TNF-α and IL-1β mRNA levels in 11 and 9 months old mice, respectively. Age dependent
amyloid plaque accumulation in APP/PS1 Tg mice was estimated by stereology in 3, 6, 9, 12, and 15 month old mice
and microglial response was investigated by immunohistochemistry for CD11b and by stereological estimation of the
number of Iba1+ cells in 4, 8 and 18 months old mice.
Stainings of CD11b+ microglia/macrophages showed an age- and transgene-dependent activation of microglia which
were observed to cluster around Aβ plaques in Tg mice. This correlated well with numbers of Iba1+ cells in young vs.
old Tg mice and wildtype (WT) mice, with an age-dependent increase in Aβ plaque burden. The mRNA levels of TNF-α
increased significantly with age in APP/PS1 Tg mice, however, we saw close to baseline levels of IL-1β mRNA. Following treatment with an endotoxin, levels of IL-1β mRNA in Tg mice rose significantly compared to PBS treated mice
(~2 fold) and also compared to WT mice receiving treatment. TNF-α mRNA levels were also increased after endotoxin
treatment but not to the same extent.
In conclusion, the results point to that the continuous Aβ deposition induces a chronic low-grade microglial-driven inflammatory reaction in the aged APP/PS1 Tg mice and that a long-term inflammatory challenge might induce and enhance
the production of potentially neurotoxic substances such as TNF-α and IL-1β.
epi-1 – a novel gene for modulation of amyloid-beta toxicity in a worm model of Alzheimer’s disease
Louise Toft Jensen and Anders Olsen
Department of Molecular Biology, Aarhus University
The nematode Caenorhabditis elegans is an excellent model for studying aging due to its short lifespan and powerful genetics. Since most nematode genes and molecular mechanisms have human homologs, results obtained in C.elegans
can often be extrapolated to humans. Using the genetic “toolbox” available in C. elegans, several elegant disease models have been established to study neurodegenerative diseases such as Alzheimer’s and Parkonson’s disease in vivo.
EPI-1 is a laminin homolog that constitutes a major component of the basal lamina. We have studied the role of EPI-1 in
a C. elegans model of Alzheimer’s disease, where human Aβ42 is expressed in the body wall muscle of the worm resulting in an age-related increase in Aβ-aggregation and paralysis. When epi-1 is knocked down using RNAi in this model,
we see a dramatically increase in paralysis compared to control RNAi treated animals. Complementing this analysis,
Western blots probed with anti-Aβ42-ab revealed large differences in the distribution of Aβ oligomers in epi-1 RNAi
treated animals compared to controls. Interestingly, epi-1 also seems to play a role in the toxicity of protein aggregation
in general, as is the case for other Aβ-toxicity modulating genes, e.g. hsf-1 and hsp-70.
A B S T R A C T S
Hippocampal Neuroplasticity in a Genetic Rat Model of Depression
Susanne Kaae1,2, Fenghua Chen1,2, Gregers Wegener2, Jens Randel Nyengaard1
1.
2.
Stereology and Electron Microscopy Lab., Aarhus University Hospital NBG
Center for Basic Psychiatric Research, Psychiatric Hospital Riiskov
Despite the fact that medical treatment of depressive disorder is possible, there is still a lack of knowledge about the
etiology and pathophysiology of the disease, and side effects and relapses are still a big problem.
Recent hypothesis have suggested that depressive disorders may be related to impairments of structural plasticity and
neural cellular resilience in hippocampus and other brain regions. The clinical effect of antidepressant treatment may be
due to alterations in neurogenesis and synaptogenesis in these regions.
The aim of the current study is to investigate whether antidepressant treatment increases the rate of neurogenesis in the
hippocampus. Two sub-studies are performed, one investigating the structural effects of imipramine (1) (published) and
the other (ongoing) investigating the effects of electroconvulsive seizure (ECS), which is considered the most effective
treatment for severe depression.
The Flinders Sensitive Line (FSL) rats were used as a genetic animal model of depression, and the Flinders Resistant
Line (FRL) rats are used as controls. FSL and FRL rats received either imipramine (15 mg/kg) or saline daily for 25 days,
or they were given ECS or sham treatment daily for 10 days. The behavioral response was measured as immobility in
the forced swim test. The volume of hippocampus as well as the number of neuron and synapse numbers in hippocampus were estimated using design based stereological methods.
Results from the imipramine study show a significantly smaller baseline level of the volume and the number of neurons
and spine synapses in the hippocampus of the untreated FSL rats, correlating to a decrease in immobility in the forced
swim test. After treatment the number of neurons in the granule cell layer (GCL) of hippocampus significantly increases
in the FSL rats and the behavior changes to significantly less immobility. Also the number of neurons in the hilus is increased, although not significant.
The number of neurons in GCL and hilus is not different comparing treated FSL rats to the untreated FRL rats.
These findings indicate that chronic imipramine treatment reverses the suppression of neurogenesis in the hippocampus of the “depressed” FSL rats, and this occurs in correlation with the behavioral response.
Results from the ECS study are still being collected, but the conclusion so far is that depressive disorders may be related
to impairments of structural plasticity in the hippocampus, and furthermore antidepressant treatment counteracts the
structural impairments.
1. Chen F, Madsen TM, Wegener G, Nyengaard JR. Imipramine treatment increases the number of hippocampal synapses and neurons in a genetic
animal model of depression. Hippocampus. 17, 2009 Nov.
P A R T I C I P A N T S
O D E N S E
N AM E
LAB
EMAIL
Trevor Owens
Owens lab, Odense University
towens@health.sdu.dk
Rebecca Radde
rebecca.radde@web.de
Reza Khorooshi
rkhorooshi@health.sdu.dk
Mohammed Salem
moalz04@student.sdu.dk
Christina Fenger
Neurobiology Research, IMM,SDU
cfenger@health.sdu.dk
Laura Ilkjaer
lilkjaer@health.sdu.dk
Morten Kobaek Larsen
mkobaek@hotmail.com
Kate Lambertsen
klambertsen@health.sdu.dk
Matilda Degn Winter
Glostrup Research Institute
matildadegn@gmail.com
Bente Finsen
bfinsen@health.sdu.dk
Ditte Elmann
dellman@health.sdu.dk
A A R H U S
N AM E
LAB
EMAIL
Aage K Olsen Alstrup
PET-centre, AU Hospital
aage@pet.auh.dk
Anne M. Landau
PET-centre & CFIN, AU Hospital
annie@pet.auh.dk
Line Gebauer
CFIN, AU
linegebauer@me.com
Mette Simonsen
PET-centre, AU Hospital
simonsen@pet.auh.dk
Arne Møller
PET-centre & CFIN, AU and University Hospital
arne@pet.auh.dk
Jørgen Scheel-Krüger
kruger@cfin.dk
Allan Rasmusson
Stereology and EM Res Lab, AU
alras@ki.au.dk
Fenghua Chen
fenghua.chen@ki.au.dk
Susanne Kaae
SUSANNEKAAE@STUDMED.
AU.DK
Anders Olsen
Dept of Molecular Biology, AU
ano@mb.au.dk
Louise T. Jensen
ltj@mb.au.dk
Kim Ryun Drasbek
ryun@mb.au.dk
Mette Berendt
Dept of Small Animal Clin Sci, LIFE, KU
mbe@life.ku.dk
Nadia Fredsø Andersen
Dept of Small Animal Clin Sci, LIFE, KU
nadiaf@life.ku.dk
Adjmal Nahim
adjmalnahimi@gmail.com
Jesper Fontain
fontain@studmed.au.dk
Anne Sabers
Epilepsy Clinic – The neuroscience Center,
Rigshospitalet
anne.sabers@rh.regionh.dk
C O P E N H A G E N
N AM E
LAB
EMAIL
Uffe Kristiansen
University of Copenhagen, FARMA
uk@farma.ku.dk
Celia Kjærby Hansen
CELH@Lundbeck.com
Mark Holm Christensen
mhch@farma.ku.dk
Charlotte Simonsen
chs@farma.ku.dk
Jessica Klein
jkl@farma.ku.dk
Kristi Kohlmeier
kak1@farma.ku.dk
Morten P. Kristensen
mpk@farma.ku.dk
Albert Gjedde
INF, University of Copenhagen
gjedde@sund.ku.dk
David Woldbye
Neuropsykiatrisk Laboratorium, INF
woldbye@sund.ku.dk
Ditte Dencker Nielsen
dittedencker@hotmail.com
Casper Goetzsche
cr@nplab.dk
Pia Weikop
piw@nplab.dk
Gunnar Soerensen
gs@nplab.dk
Zindy Raida
University of Copenhagen, INF
zindy@sund.ku.dk
Bente Pakkenberg
Research Laboratory for Stereology and Neuroscience
bentepakkenberg@hotmail.
com
Katrine Fabricius
katrinefabricius@hotmail.com
S. Kaalund
sanne.kaalund@gmail.com
N. Eriksen
nina.eriksen1@gmail.com
Anna Karlsen
annakarlsen@gmail.com
J. Riise
drivmiddel@hotmail.com
Flemming Fryd Johansen
University of Copenhagen, BRIC
flemming.johansen@bric.dk
Mark Holm Christensen
mach@farma.ku.dk
Kim Anker Kristiansen
Research unit, Glostrup Hospital
KIANKR01@glo.regionh.dk
Rune Skovgaard
University of Copenhagen
RSR@sund.ku.dk
Henrik Hasseldam
henrik.hasseldam@bric.dk
Ida S. Rasmussen
University of Copenhagen, SUND
idasr.patologi@gmail.com
Anders Hay-Schmidt
C O P E N H A G E N
N AM E
LAB
EMAIL
Issazadeh Shohreh
shohreh.issazadeh@bric.dk
Christian Berg
christian.berg@bric.dk
Martin Kuss
martin.kuss@bric.dk
Jeanette Svensson
jeanette.svensson@bric.ku.dk
Yawei Liu
yawei.liu@bric.dk
Hilmar van Weering
hilmar.vanweering@bric.ku.dk
Kim Noerring
kim.noerring@bric.ku.dk
Robert Bockermann
Robert.bockermann@bric.
ku.dk
P A T I E N T
Patienthotellet
Afsnit 52
indgang 52
Blegdamsvej 9
2100 København Ø
H O T E L
G A L A
D I N N E R
Friday 11 th of June 2010 at 20:00 the OAK Meeting speakers and participants will have a gala dinner.
The dinner will take place at restaurant
Romarin - where we look forward to spend a nice evening with you.
Romarin is located at:
Ryegade 90, 2100 København
B R I C
BRIC is situated at Ole Maaløes vej 5 (A on map)
Bus 6A goes from Nørreport to BRIC
N O T E S

Here - OAK

Transcription

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