This year we have collected 58 brains? ? To date we have collected: 1,017 brains? ? Diagnosis: Schizophrenia 147; Bipolar 40; Major Depressive Disorder 76; Normal Control 271; Affective Disorder 49; Substance Abuse 161; Fetal Brain 44; Others 171; and 58 to be characterized? ? The major focus of our studies is schizophrenia, a syndrome involving cognitive deficits, negative symptoms, and psychosis, usually starting in late adolescence or early adulthood. The recent discovery of variations (single nucleotide polymorphisms (SNPs)) in a number of genes associated with schizophrenia not only speaks to a significant component of the etiology, but allows for increased understanding of the neuropsychology, neuroimaging, and neuropathology of the disorder. Postmortem human brain studies guided by genetic advances are a valuable, if not essential component, in elucidating the cellular and molecular pathophysiology of schizophrenia. Moreover, improved understanding of a genetic component in schizophrenia may enhance diagnostic abilities and, hopefully, lead to new treatments. ? Postmortem human brain studies are limited by the quality and quantity of the specimens. ? ? Brain Procurement: Although the number of brains collected per year has remained relatively constant for the past ten years, we have become increasingly selective over the past four years in choosing our cases. Moreover, the entire collection has undergone a case-by-case review leading to a reduction in the number of brains for study. These selected cases all have informed consent from the next of kin at the time of donation, many with extensive medical records available for review. In addition, whenever possible, we have conducted interviews with the next of kin for additional clinical information. The tissue itself is screened for macro- and microscopic neuropathology, and multiple screens are performed for RNA and protein integrity. Toxicological screens are conducted for substances of abuse, as well as for prescribed psychotropic medications. ? ? Clinical Review and Characterization: Informed consent for research on the brain and related tissue samples from the next of kin is obtained at or before the time of autopsy including the brain, blood, and a hair specimen (for toxicology) as well as permission to obtain medical records. Next of kin are interviewed with a 14-item telephone-screening questionnaire. The next of kin are advised that they will receive a follow-up interview in greater depth within several months. This in-depth interview relies upon three screening devices: a SCID (First et al., 1997), a modified psychological autopsy (Kelly and Mann, 1996) and a modified version of the DEAD (Diagnostic Evaluation after Death) (Zalcman et al., 1983) in order to collect relevant clinical information from the next of kin in order to enhance our diagnostic classification.? ? The medical examiners offices provide demographic information (age, gender, race, etc.), a police report, interviews with the next of kin, and/or relevant medical personnel and witnesses, toxicology screens, and a general autopsy report that includes the cause and manner of death. The medical examiners offices perform toxicology screens in the vast majority of cases. If toxicology screens are not performed we have contracted a laboratory to complete the screens.? ? An extensive review of clinical records and all other available information is conducted by trained personnel in the Section on Neuropathology, and then independently reviewed by two board certified psychiatrists. If the latter agree on diagnosis, the case is then formally reviewed at a diagnostic conference attended by the two psychiatrists, our neuropathologist, the clinical interviewing team, and the data base specialist where a DSM-IVR (APA, 2000) final diagnosis is recorded. In the event that the two psychiatrists disagree, a third board certified psychiatrist reviews the case independently prior to the diagnostic conference. No final DSM-IVR diagnosis is assigned at the diagnostic conference unless two psychiatrists concur in the diagnoses. Similar rigor is used in designating normal control subjects who must have no history of psychiatric or neurological disorders, no history of significant substance abuse or dependence, a negative toxicology screen at autopsy, and a neuropathological screen free from abnormalities. Nicotine use has been carefully reviewed and is not an exclusionary criterion for normal control subjects.? ? The neuropathological screening involves two components, a macroscopic examination and a microscopic inspection. A board certified neuropathologist conducts the macroscopic examination at the time of tissue collection, weighing and inspecting the whole brain, upper cervical spinal cord, pituitary and pineal glands, dura, and intracranial vasculature. The microscopic examination involves sectioning and staining tissue from the frontal, temporal, and occipital poles, the posterior hippocampus, and the cerebellar vermis, looking for evidence of neurodegeneration or other pathological processes. Sections are routinely stained to reveal neurofibrillary tangles and neuritic plaques that are counted in order to apply Khachaturian criteria to establish the neuropathological diagnosis of Alzheimers disease (Khachaturian, 1985). In addition, sections are taken from any region that shows signs of macroscopic pathology, such as an apparent infarct or abscess, for additional neuropathological analysis. A formal macroscopic and microscopic neuropathology report is written from the information obtained from these examinations.? ? Screening for Molecular Biology Studies: The following screening procedures have been performed on every brain prior to inclusion in our studies:? a. Measurement of tissue pH from homogenized cerebellar hemispheric tissue.? b. Measurement of mRNA expression of a panel of putatively constitutively expressed genes (such as actin and cyclophilin) using in situ hybridization histochemistry on 14 micron cerebellar hemispheric sections.? c. Extraction of total RNA by Qiagen and separation of total RNA using agarose gels to qualitatively determine the integrity and intensity of the 28S and 18S ribosomal RNA bands.? d. Quantitative analysis of total RNA integrity and 28S to 18S ratios using capillary electrophoresis on a Bioanalyzer 2100 (Agilent, Inc.).? e. Acceptable postmortem intervals (PMI) in human brain for each protein studied with Westerns or ELISAs are inferred from rat studies investigating the PMI when protein levels for that protein significantly decline (Halim et al., 2003).? f.. Similarly, in order to determine the effects of medications in the human studies of mRNA and protein expression, we are using brain tissue from rats chronically treated with various doses of antipsychotic drugs (haloperidol and clozapine) (Lipska et al., 2001, 2003).? g. From these screening measurements, we have developed exclusionary criteria for the use of human brain tissue in our experiments: tissue pH < 6.00, PMI > 60 hours; abnormal cerebellar actin and cyclophilin expression levels in cerebellar in situ hybridization (greater than two standard deviations from the mean); and a 28S/18S ribosomal RNA ratio < 1.2.? h. In so far as the vast majority of our cases (over 90%) come from medical examiner offices, few, if any, of our brains come from patients who have died with prolonged agonal states. The latter is clearly associated with low pH and increased RNA degradation (Tomita et al., 2004).
| Kaalund, S S; Newburn, E N; Ye, T et al. (2014) Contrasting changes in DRD1 and DRD2 splice variant expression in schizophrenia and affective disorders, and associations with SNPs in postmortem brain. Mol Psychiatry 19:1258-66 |
| Bigos, Kristin L; Mattay, Venkata S; Callicott, Joseph H et al. (2010) Genetic variation in CACNA1C affects brain circuitries related to mental illness. Arch Gen Psychiatry 67:939-45 |
