Determination of Reverse Transcriptase (RT) Activity in Clinical and Post-Mortem Samples From Individuals with Schizophrenia and Bipolar Disorder

DETERMINATION OF REVERSE TRANSCRIPTASE (RT)

ACTIVITY IN CLINICAL AND POST-MORTEM SAMPLES OBTAINED FROM

INDIVIDUALS WITH SCHIZOPHRENIA AND BIPOLAR DISORDER

F. Yee*, L.

Jones-Brando, E.F. Torrey, R.H. Yolken and the Stanley

Neuropathology Consortium. Stanley Division of Developmental

Neurovirology, Johns Hopkins University School of Medicine,

Baltimore, MD

Our laboratory is interested

in studying the role of retroviruses in the etiology of

neuropsychiatric illnesses such as schizophrenia and bipolar

disorder. Retroviruses may represent an important link

between genetic and environmental factors, as this would

account for horizontal and vertical transmission, and these

viruses may be involved with the disease process in

subpopulations affected with these serious mental illnesses.

An important component of infectious retroviruses is that

they contain the reverse transcriptase (RT) enzyme, and

consequently they can be detected by assays for RT activity.

In this study, we set out to

examine the levels of RT activity in various samples, e.g.

cerebrospinal fluid (CSF) from first episode patients with

schizophrenia, post-mortem cisternal fluids and various brain

regions from individuals with schizophrenia, bipolar

disorder, depression without psychosis, as well as unaffected

individuals. Before proceeding with these experiments, it is

important to determine the baseline RT activity present in

these samples, which may be due to the presence of endogenous

retroviruses. We have modified an ultrasensitive RT assay

called product-enhanced RT (PERT; Pyra el al., PNAS

91:1544-1548, 1994), by adding a second round of polymerase

chain reaction (PCR) amplification. When the resulting PCR

products are visualized with the nucleic acid dye SYBR-Green

(Molecular Probes) and analyzed with the Fluorimager Sl

(Molecular Dynamics) we are able to detect as little as 10-7

units of Moloney murine leukemia virus-reverse transcriptase

(MMLV-RT) activity in the PERT assay. Our preliminary PERT

data indicate that there is a non-significant trend of

increased RT activity in the cerebellum of groups with

schizophrenia (n=11) and bipolar disorder (n=13) compared

with the non-psychotic depression (n=12) and unaffected

(n=11) groups.