- Email: [email protected]
PROTEOMIC AND WESTERN BLOT ANALYSIS OF HIPPOCAMPAL SUBREGIONS IN SCHIZOPHRENIA AND BIPOLAR DISORDER
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Abstracts / Schizophrenia Research 102/1–3, Supplement 2 (2008) 1–279
Methods: Dorsolateral prefrontal grey and white matter was obtained from the Stanley Medical Research Institute Brain Collection (schizophrenia, bipolar disorder and controls; all n=35). Cholesterol levels were quantified using HPLC with ELSD. DNA was extracted and ApoE genotyping performed using PCR-RFLP. Results: In grey matter, cholesterol levels were 10% and 6% lower in schizophrenia and bipolar disorder respectively, relative to the control group. In white matter cholesterol levels were unchanged in both disorders. Repeated measures ANOVA revealed an effect of region, but no significant effect of diagnosis or region by diagnosis interaction. Preliminary analyses of the first 60 cases indicate that overall e4 alleles are associated with lower cholesterol levels in white but not grey matter. Conclusions: We were not able to identify deficits in cholesterol in the dorsolateral prefrontal region in schizophrenia or bipolar disorder. ApoE genotype may influence regulation of brain cholesterol. References [1] Beasley CL et al (2005). Reductions in cholesterol and synaptic markers in association cortex in mood disorders. [2] Bipolar Disord. 7:449-455.
495 – PROTEOMIC AND WESTERN BLOT ANALYSIS OF HIPPOCAMPAL SUBREGIONS IN SCHIZOPHRENIA AND BIPOLAR DISORDER Melanie Focking 1 , Renata Schoeman 2 , Patrick Dicker 3 , Lance Hudson 1 , Michael J Dunn 4 , David R Cotter 1 1 Department of Psychiatry, Royal College of Surgeons in Ireland, Dublin; 2 Department of Psychiatry, University of Stellenbosch, Cape Town; 3 Department of Molecular and Cellular Therapeutics, Royal College of Surgeons in, Dublin; 4 CIPRC, University College Dublin, Dublin [email protected] Introduction: The hippocampus has critical roles in learning and memory and is centrally implicated in many neuropsychiatric disorders. It is divided into subregions, and these differ with regards to morphology, connectivity, electrophysiology and susceptibility to insults. There is evidence that hippocampal changes are amongst the central components of schizophrenia and precede the onset of the illness and that changes involve the hippocampal subregions differentially. These different subregions have distinct roles in regulation of hippocampal circuitry and alterations within them are likely to contribute in a primary way to the clinical presentation. In a non-hypothesis driven approach we aim to characterize the differential protein expression in each of the hippocampal subregions in schizophrenia and bipolar disorder compared to control tissue. Methods: We used laser-assisted microdissection, and Difference-inGel-Electrophoresis to enrich for these tissues and to compare protein profiles. Image analysis was carried out using Progenesis-SameSpots. Samples were grouped according to the different disease/control groups. Results: We found 213 spots to be differentially expressed within both diseases and over all four subgroups. Spots are currently being identified by mass spectrometry. In a different approach, hypothesizing that glutamatergic synapses, particularly those regulated by the N-methyl-D-aspartate receptor (NMDAR), play a central role in the disease we have assessed proteins in the hippocampus (CA2/3) of schizophrenic patients, using 2D Western blots. We assessed the differential hippocampal expression of NR3A, NF-L, PACSIN-1, DYN-1 and septin-5. Conclusions: We hypothesize that differential expression of these proteins will be observed in the hippocampus of schizophrenia. This study will provide novel data clarifying the nature of the NMDA-receptor related pathology in schizophrenia. References [1] Pennington K et al., Mol Psychiatry 2007 Oct: Epub ahead of print
496 – THE SANT JOAN DE DEU BRAIN BANK IN SCHIZOPHRENIA RESEARCH Josep Maria Haro 1 , Mercedes Roca 2 , Ana Escanilla 2 , Alfonso Monge 2 , Vicente Baño 2 , Lluis Maria Planchat 3 , Juan Costa 2 1 Fundacio Sant Joan de Déu, Esplugues de Ll. (Barcelona); 2 Sant Joan de Deú-SSM, Sant Boi de Llobregat; 3 Institut Anatomic Forense, Sant Boi de Llobregat, Spain [email protected] Introduction: Central Nervous System Tissue Banks (CNSTB, Brain Banks) are collections of samples of brains and other neurological tissues from donors who are either healthy or affected by some sort of neurological or mental illness. Brain banks are useful in the analyses of the genomic and proteomic analysis of neuropsychiatric diseases. Methods: The Sant Joan de Déu Brain Bank (SJDBB) was created in 2004 and is an archive of tissue samples affected by mental illnesses. Donors, or their legal representatives, provide informed consent, and relatives are also informed. A complete psychiatric and physical examination is carried out with all donors. Diagnoses are confirmed by means of the Structured Clinical Interview (SCID) for DSM-IV, and the both severity of symptoms and neuropsychological functioning are assessed. Samples are processed following the protocols established by Brain Net Europe. Results: The number of donors currently stands at 105. Of the donors, 23 have died, providing 23 frozen samples of CSF, cerebral sections. Although all cases had a primary diagnosis of psychotic disorder, the first cases have a high prevalence of neurodegenerative diseases and vascular diseases, which is not infrequent in newly created brain banks. Conclusions: This brain bank opens the opportunity to conduct research on ageing in psychiatric patients, the co-morbidity between mental and neurological disorders and the possible role of treatments and lifestyles in cardiovascular diseases. References [1] Garrick, T., Howell, S., Terwee, P., Redenbach, J., Blake, H., & Harper, C. 2006, "Brain donation for research: who donates and why?", J. Clin. Neurosci., vol. 13, no. 5, pp. 524-528. [2] Bell, J.E., Bogdanovic, N., Bruck, W., Budka, H., Ferrer, I., Giaccone, G., Kovacs, G.G., Meyronet, D., Palkovits, M., Parchi, P., Patsouris, E., Ravid, R., Reynolds, R., Riederer, P., Roggendorf, W., Schwalber, A., Seilhean, D., & Kretzschmar, H. 2007, "How a neuropsychiatric brain bank should be run: a consensus paper of Brainnet Europe II", J. Neural Transm., vol. 114, no. 5, pp. 527-537.
497 – EXPRESSION OF DOUBLECORTIN DURING HUMAN CORTICAL DEVELOPMENT Sinthuja Sivagnanasundaram 1 , Sabine Bahn 2 , Michael Elashoff 3 , Maree Webster 4 , Cynthia Weickert 5 1 Prince of Wales Medical Research Institute, Sydney; 2 University of Cambridge, Cambridge; 3 Cardiodx, California; 4 Stanley Medical Research Institute, Maryland; 5 Schizophrenia Research Institute, University of New South Wales, POWMRI, Sydney, Australia [email protected] Introduction: Altered neuronal density in the white matter of the prefrontal cortex (PFC) of schizophrenia patients [1] has implicated abnormal neuronal migration in early brain development as a possible mechanism in the pathogenesis of schizophrenia. Doublecortin (DCX), highly expressed in migrating and differentiating neurons [2,3], is involved in neuronal migration [4]. As an initial step in our investigation of the possible role of DCX in the pathogenesis of schizophrenia we have examined the levels of DCX mRNA in the PFC of the developing human brain. Methods: Microarray methodology was used to assess DCX mRNA levels between developmental time points of 6 weeks to 49 years. Total RNA was extracted from the PFC of 45 individuals and hybridized to HG-U133 version 2.0+ GeneChips (Affymetrix CA, USA).
Abstracts / Schizophrenia Research 102/1–3, Supplement 2 (2008) 1–279
Methods: Dorsolateral prefrontal grey and white matter was obtained from the Stanley Medical Research Institute Brain Collection (schizophrenia, bipolar disorder and controls; all n=35). Cholesterol levels were quantified using HPLC with ELSD. DNA was extracted and ApoE genotyping performed using PCR-RFLP. Results: In grey matter, cholesterol levels were 10% and 6% lower in schizophrenia and bipolar disorder respectively, relative to the control group. In white matter cholesterol levels were unchanged in both disorders. Repeated measures ANOVA revealed an effect of region, but no significant effect of diagnosis or region by diagnosis interaction. Preliminary analyses of the first 60 cases indicate that overall e4 alleles are associated with lower cholesterol levels in white but not grey matter. Conclusions: We were not able to identify deficits in cholesterol in the dorsolateral prefrontal region in schizophrenia or bipolar disorder. ApoE genotype may influence regulation of brain cholesterol. References [1] Beasley CL et al (2005). Reductions in cholesterol and synaptic markers in association cortex in mood disorders. [2] Bipolar Disord. 7:449-455.
495 – PROTEOMIC AND WESTERN BLOT ANALYSIS OF HIPPOCAMPAL SUBREGIONS IN SCHIZOPHRENIA AND BIPOLAR DISORDER Melanie Focking 1 , Renata Schoeman 2 , Patrick Dicker 3 , Lance Hudson 1 , Michael J Dunn 4 , David R Cotter 1 1 Department of Psychiatry, Royal College of Surgeons in Ireland, Dublin; 2 Department of Psychiatry, University of Stellenbosch, Cape Town; 3 Department of Molecular and Cellular Therapeutics, Royal College of Surgeons in, Dublin; 4 CIPRC, University College Dublin, Dublin [email protected] Introduction: The hippocampus has critical roles in learning and memory and is centrally implicated in many neuropsychiatric disorders. It is divided into subregions, and these differ with regards to morphology, connectivity, electrophysiology and susceptibility to insults. There is evidence that hippocampal changes are amongst the central components of schizophrenia and precede the onset of the illness and that changes involve the hippocampal subregions differentially. These different subregions have distinct roles in regulation of hippocampal circuitry and alterations within them are likely to contribute in a primary way to the clinical presentation. In a non-hypothesis driven approach we aim to characterize the differential protein expression in each of the hippocampal subregions in schizophrenia and bipolar disorder compared to control tissue. Methods: We used laser-assisted microdissection, and Difference-inGel-Electrophoresis to enrich for these tissues and to compare protein profiles. Image analysis was carried out using Progenesis-SameSpots. Samples were grouped according to the different disease/control groups. Results: We found 213 spots to be differentially expressed within both diseases and over all four subgroups. Spots are currently being identified by mass spectrometry. In a different approach, hypothesizing that glutamatergic synapses, particularly those regulated by the N-methyl-D-aspartate receptor (NMDAR), play a central role in the disease we have assessed proteins in the hippocampus (CA2/3) of schizophrenic patients, using 2D Western blots. We assessed the differential hippocampal expression of NR3A, NF-L, PACSIN-1, DYN-1 and septin-5. Conclusions: We hypothesize that differential expression of these proteins will be observed in the hippocampus of schizophrenia. This study will provide novel data clarifying the nature of the NMDA-receptor related pathology in schizophrenia. References [1] Pennington K et al., Mol Psychiatry 2007 Oct: Epub ahead of print
496 – THE SANT JOAN DE DEU BRAIN BANK IN SCHIZOPHRENIA RESEARCH Josep Maria Haro 1 , Mercedes Roca 2 , Ana Escanilla 2 , Alfonso Monge 2 , Vicente Baño 2 , Lluis Maria Planchat 3 , Juan Costa 2 1 Fundacio Sant Joan de Déu, Esplugues de Ll. (Barcelona); 2 Sant Joan de Deú-SSM, Sant Boi de Llobregat; 3 Institut Anatomic Forense, Sant Boi de Llobregat, Spain [email protected] Introduction: Central Nervous System Tissue Banks (CNSTB, Brain Banks) are collections of samples of brains and other neurological tissues from donors who are either healthy or affected by some sort of neurological or mental illness. Brain banks are useful in the analyses of the genomic and proteomic analysis of neuropsychiatric diseases. Methods: The Sant Joan de Déu Brain Bank (SJDBB) was created in 2004 and is an archive of tissue samples affected by mental illnesses. Donors, or their legal representatives, provide informed consent, and relatives are also informed. A complete psychiatric and physical examination is carried out with all donors. Diagnoses are confirmed by means of the Structured Clinical Interview (SCID) for DSM-IV, and the both severity of symptoms and neuropsychological functioning are assessed. Samples are processed following the protocols established by Brain Net Europe. Results: The number of donors currently stands at 105. Of the donors, 23 have died, providing 23 frozen samples of CSF, cerebral sections. Although all cases had a primary diagnosis of psychotic disorder, the first cases have a high prevalence of neurodegenerative diseases and vascular diseases, which is not infrequent in newly created brain banks. Conclusions: This brain bank opens the opportunity to conduct research on ageing in psychiatric patients, the co-morbidity between mental and neurological disorders and the possible role of treatments and lifestyles in cardiovascular diseases. References [1] Garrick, T., Howell, S., Terwee, P., Redenbach, J., Blake, H., & Harper, C. 2006, "Brain donation for research: who donates and why?", J. Clin. Neurosci., vol. 13, no. 5, pp. 524-528. [2] Bell, J.E., Bogdanovic, N., Bruck, W., Budka, H., Ferrer, I., Giaccone, G., Kovacs, G.G., Meyronet, D., Palkovits, M., Parchi, P., Patsouris, E., Ravid, R., Reynolds, R., Riederer, P., Roggendorf, W., Schwalber, A., Seilhean, D., & Kretzschmar, H. 2007, "How a neuropsychiatric brain bank should be run: a consensus paper of Brainnet Europe II", J. Neural Transm., vol. 114, no. 5, pp. 527-537.
497 – EXPRESSION OF DOUBLECORTIN DURING HUMAN CORTICAL DEVELOPMENT Sinthuja Sivagnanasundaram 1 , Sabine Bahn 2 , Michael Elashoff 3 , Maree Webster 4 , Cynthia Weickert 5 1 Prince of Wales Medical Research Institute, Sydney; 2 University of Cambridge, Cambridge; 3 Cardiodx, California; 4 Stanley Medical Research Institute, Maryland; 5 Schizophrenia Research Institute, University of New South Wales, POWMRI, Sydney, Australia [email protected] Introduction: Altered neuronal density in the white matter of the prefrontal cortex (PFC) of schizophrenia patients [1] has implicated abnormal neuronal migration in early brain development as a possible mechanism in the pathogenesis of schizophrenia. Doublecortin (DCX), highly expressed in migrating and differentiating neurons [2,3], is involved in neuronal migration [4]. As an initial step in our investigation of the possible role of DCX in the pathogenesis of schizophrenia we have examined the levels of DCX mRNA in the PFC of the developing human brain. Methods: Microarray methodology was used to assess DCX mRNA levels between developmental time points of 6 weeks to 49 years. Total RNA was extracted from the PFC of 45 individuals and hybridized to HG-U133 version 2.0+ GeneChips (Affymetrix CA, USA).