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Genetic and experiential influences on CSF monoamine metabolites in a primate model of temperament and psychopathology
46A
BIOL PSYCHIATRY 1989;25:144A-148A
Genetics
from embryonic mice telencephalons at different stages of development and performing subtractive hybridizations between these libraries in order to isolate genes that are expressed at specific times of development. Using this paradigm we expect to isolate genes that are involved in controlling cell migration, cell fate, axon pathway choice, and synapse formation. We are performing the subtractive hybridization using specially designed vectors. The basis for the technique is asymmetric insertion of cDNA into double-stranded plasmids which have the capability of also growing in a single-stranded form. The single-stranded form of the DNA will be used in the hybridization. These experiments will serve as an early step in our understanding of the genes which regulate development of the brain, and should eventually aid in the understanding of such complex developmental disorders as autism.
294 GENE TRANSFER AND EXPRESSION IN MAMMALIAN CELL LINES: A METHOD TO STUDY REGULATION OF NEUROPEPTIDE PROCESSING James H. Meador-Woodruff, Benedetto Pellerito, Mary Hoversten, Donald Martin, Huda Akil Ann Arbor,
MI
Many neuropeptides undergo tissue-specific processing, resulting in the production of various bioactive domains. Brain region-specific processing differences may have physiological relevance, due to differences in bioactivity of the proteolytic products. The regulation of the processing of neuropeptides has been difficult to study in the brain, however, due in part to extremely low concentrations of peptides and processing enzymes, regional heterogeneity, and the complexity of neuronal circuitry and feedback. One powerful tool that has recently become available to study cellular regulatory mechanisms underlying cell-specific processing differences is gene transfer and expression in cultured clonal cell lines. We have begun to study the regulation of posttranslational modification of neuropeptides using this technique, concentrating on the ACTH/beta-endorphin precursor, proopiomelanocortin (POMC). We have constructed an expression vector containing monkey POMC cDNA, and have propagated multiple stable transfected lines, including neuronal, glial, and endocrine cell types. These cell lines are able to express POMC, and some are able to process this propeptide into functional domains. The development of these cell lines transfected with POMC (or other) cDNA will allow us to address questions of regulation of cell-specific processing, utilizing techniques such as site-directed mutagenesis.
295 GENETIC AND EXPERIENTIAL INFLUENCES ON CSF MONOAMINE METABOLITES IN A PRIMATE MODEL OF TEMPERAMENT AND PSYCHOPATHOLOGY J.D. Higley, M. Linnoila, S.J. Suomi Bethesda,
MD
Studies of nonhuman primates have shown that individual differences (ID) in the personality trait reactivity (timidity, behavioral inhibition, etc.) are related to anxiety and alcohol consumption. This trait is exacerbated by early developmental experiences such as peer-only rearing, and is heritable. This paper is an investigation of the
BIOL PSYCHIATRY 1989;25:144A-148A
Genetics
147A
contributions of CNS systems to ID in reactivity, assessed via CSF monoamine metabolites. Twenty Rhesus monkeys were reared for their first six months either with only peers (N = 8) or as controls with their mothers (N = 12). When subjects were six and 18 months of age, they underwent a series of four 4-day social separations. Thereafter, both groups were placed together and received identical treatment. In both years, prior to and during separations, CSF was obtained via cistemal puncture. The more reactive peer-only reared subjects had higher levels of MHPG across both years and in the second year, higher levels of 5-HIAA. Within group ID were remarkably stable: Mean separation levels of MHPG, HVA, and 5-HIAA correlated at r = .47, .71, and .5 1 respectively. When these subjects and an additional 20 were grouped according to their unfamiliar biological sire, CSF 5-HIAA and HVA concentration were heritable.
296 Xq28 DNA MARKERS AND INHERITANCE OF MANICDEPRESSIVE ILLNESS: REJECTION OF LINKAGE Wade Berrettini, Elliot Gershon, Lynn Goldin Bethesda, MD
Close linkage of manic-depressive illness (MDI) to the classical Xq28 markers, color blindness (CB), and glucose-6-phosphate dehydrogenase (G6PD) deficiency, has been reported (Baron et al, Nature 326:289). We attempted to confirm these results in nine pedigrees (in which there was no male-to-male transmission of any form of MDI), using two Xq28 DNA markers (DXS52 and DSX13) which are tightly linked (within 3 CM) to the CB and G6PD loci. Assumptions concerning disease classification, mode of inheritance and gene frequency were similar to those of Baron et al. Linkage could be rejected (LOD score < - 2) up to 20 CM from the DXS52 locus with a LOD score < - 9 at that locus. When these nine families are combined with our previous series of 5 MD1 pedigrees (Arch Gen Psychiatry 36: 1423), linkage would have been detected if only 25% of the families studied were linked. We are unable to reconcile these results with previous reports that Xq28-linkage in MD1 is a common phenomenon.
297 ADRENERGIC RECEPTOR GENES AND THEIR ROLE IN MANIC-DEPRESSIVE ILLNESS Margaret R. Hoehe, Wade H. Berrettini, Elliot S. Gershon Bethesda, MD
Several lines of evidence suggest that an abnormality of alpha (AAR) and/or beta adrenergic receptor (BAR) function may be involved in a genetic pathophysiologic vulnerability to manic-depressive illness (MDI). Using molecular genetic techniques we studied the genes encoding AARs and BARS for polymorphisms and linkage to MD1 in bipolar pedigrees. LOD scores were calculated assuming various disease classifications and various models of inheritance and a gene frequency of 1%. LOD scores for a DRA I RFLP of the A2AR located at 1Oq 24 ranged between - 3 and - 1 under various models. Further investigations with this A2AR as well as with the A2AR located on chromosome 4 are being performed. LOD scores for a BGL I RFLP of the B 1AR (1Oq 24) and for a BAN I RFLP of the B2AR (5q 32) excluded linkage for most models of transmission at theta = 0. The present results exclude abnormalities in the coding regions and their adjacent regulatory sequences of these genes as being
BIOL PSYCHIATRY 1989;25:144A-148A
Genetics
from embryonic mice telencephalons at different stages of development and performing subtractive hybridizations between these libraries in order to isolate genes that are expressed at specific times of development. Using this paradigm we expect to isolate genes that are involved in controlling cell migration, cell fate, axon pathway choice, and synapse formation. We are performing the subtractive hybridization using specially designed vectors. The basis for the technique is asymmetric insertion of cDNA into double-stranded plasmids which have the capability of also growing in a single-stranded form. The single-stranded form of the DNA will be used in the hybridization. These experiments will serve as an early step in our understanding of the genes which regulate development of the brain, and should eventually aid in the understanding of such complex developmental disorders as autism.
294 GENE TRANSFER AND EXPRESSION IN MAMMALIAN CELL LINES: A METHOD TO STUDY REGULATION OF NEUROPEPTIDE PROCESSING James H. Meador-Woodruff, Benedetto Pellerito, Mary Hoversten, Donald Martin, Huda Akil Ann Arbor,
MI
Many neuropeptides undergo tissue-specific processing, resulting in the production of various bioactive domains. Brain region-specific processing differences may have physiological relevance, due to differences in bioactivity of the proteolytic products. The regulation of the processing of neuropeptides has been difficult to study in the brain, however, due in part to extremely low concentrations of peptides and processing enzymes, regional heterogeneity, and the complexity of neuronal circuitry and feedback. One powerful tool that has recently become available to study cellular regulatory mechanisms underlying cell-specific processing differences is gene transfer and expression in cultured clonal cell lines. We have begun to study the regulation of posttranslational modification of neuropeptides using this technique, concentrating on the ACTH/beta-endorphin precursor, proopiomelanocortin (POMC). We have constructed an expression vector containing monkey POMC cDNA, and have propagated multiple stable transfected lines, including neuronal, glial, and endocrine cell types. These cell lines are able to express POMC, and some are able to process this propeptide into functional domains. The development of these cell lines transfected with POMC (or other) cDNA will allow us to address questions of regulation of cell-specific processing, utilizing techniques such as site-directed mutagenesis.
295 GENETIC AND EXPERIENTIAL INFLUENCES ON CSF MONOAMINE METABOLITES IN A PRIMATE MODEL OF TEMPERAMENT AND PSYCHOPATHOLOGY J.D. Higley, M. Linnoila, S.J. Suomi Bethesda,
MD
Studies of nonhuman primates have shown that individual differences (ID) in the personality trait reactivity (timidity, behavioral inhibition, etc.) are related to anxiety and alcohol consumption. This trait is exacerbated by early developmental experiences such as peer-only rearing, and is heritable. This paper is an investigation of the
BIOL PSYCHIATRY 1989;25:144A-148A
Genetics
147A
contributions of CNS systems to ID in reactivity, assessed via CSF monoamine metabolites. Twenty Rhesus monkeys were reared for their first six months either with only peers (N = 8) or as controls with their mothers (N = 12). When subjects were six and 18 months of age, they underwent a series of four 4-day social separations. Thereafter, both groups were placed together and received identical treatment. In both years, prior to and during separations, CSF was obtained via cistemal puncture. The more reactive peer-only reared subjects had higher levels of MHPG across both years and in the second year, higher levels of 5-HIAA. Within group ID were remarkably stable: Mean separation levels of MHPG, HVA, and 5-HIAA correlated at r = .47, .71, and .5 1 respectively. When these subjects and an additional 20 were grouped according to their unfamiliar biological sire, CSF 5-HIAA and HVA concentration were heritable.
296 Xq28 DNA MARKERS AND INHERITANCE OF MANICDEPRESSIVE ILLNESS: REJECTION OF LINKAGE Wade Berrettini, Elliot Gershon, Lynn Goldin Bethesda, MD
Close linkage of manic-depressive illness (MDI) to the classical Xq28 markers, color blindness (CB), and glucose-6-phosphate dehydrogenase (G6PD) deficiency, has been reported (Baron et al, Nature 326:289). We attempted to confirm these results in nine pedigrees (in which there was no male-to-male transmission of any form of MDI), using two Xq28 DNA markers (DXS52 and DSX13) which are tightly linked (within 3 CM) to the CB and G6PD loci. Assumptions concerning disease classification, mode of inheritance and gene frequency were similar to those of Baron et al. Linkage could be rejected (LOD score < - 2) up to 20 CM from the DXS52 locus with a LOD score < - 9 at that locus. When these nine families are combined with our previous series of 5 MD1 pedigrees (Arch Gen Psychiatry 36: 1423), linkage would have been detected if only 25% of the families studied were linked. We are unable to reconcile these results with previous reports that Xq28-linkage in MD1 is a common phenomenon.
297 ADRENERGIC RECEPTOR GENES AND THEIR ROLE IN MANIC-DEPRESSIVE ILLNESS Margaret R. Hoehe, Wade H. Berrettini, Elliot S. Gershon Bethesda, MD
Several lines of evidence suggest that an abnormality of alpha (AAR) and/or beta adrenergic receptor (BAR) function may be involved in a genetic pathophysiologic vulnerability to manic-depressive illness (MDI). Using molecular genetic techniques we studied the genes encoding AARs and BARS for polymorphisms and linkage to MD1 in bipolar pedigrees. LOD scores were calculated assuming various disease classifications and various models of inheritance and a gene frequency of 1%. LOD scores for a DRA I RFLP of the A2AR located at 1Oq 24 ranged between - 3 and - 1 under various models. Further investigations with this A2AR as well as with the A2AR located on chromosome 4 are being performed. LOD scores for a BGL I RFLP of the B 1AR (1Oq 24) and for a BAN I RFLP of the B2AR (5q 32) excluded linkage for most models of transmission at theta = 0. The present results exclude abnormalities in the coding regions and their adjacent regulatory sequences of these genes as being