Roles of the amygdala and bed nucleus of the stria terminalis in fear and anxiety measured with the acoustic startle reflex. Possible relevance to PTSD.
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Biomedical subjects
Publications and source records attributed to Y Lee.
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Bcl-2 is an oncogene that confers deregulated growth potential to B lymphocytes through its ability to inhibit apoptotic cell death. A specific molecular activity for the Bcl-2 protein has not been identified, but several lines of evidence have supported a role in protection of cells from oxidative stress. We investigated whether there is a correlation between expression of high levels of Bcl-2 and susceptibility of human Burkitt's lymphoma cell lines to H2O2-induced killing. The amount of H2O2 required to kill 50% of cells in 24 hours varied widely in the seven different lymphoma cell lines that were tested, ranging from 35 to 500 micromol/L H2O2. However, expression of high levels of endogenous Bcl-2 did not protect the cells from H2O2-induced killing, even though it was effective in protecting the cells from apoptosis induced by agents such as A23187. Thus, Bcl-2 was functional in preventing apoptosis but did not act in an antioxidant capacity. The results were confirmed using a Burkitt's lymphoma cell line overexpressing transfected bcl-2. The results may be explained by the observation that H2O2 was inefficient at inducing apoptosis in these mature B-cell lines. Nonapoptotic death induced by H2O2 was not prevented by Bcl-2.
The maturation of the ribosomal 5 S RNA in Saccharomyces cerevisiae is examined based on the expression of mutant 5 S rRNA genes, in vivo, and a parallel analysis of RNA processing, in vitro. Both types of analysis indicate that 5 S rRNA processing is not dependent on the nucleotide sequence of either the external transcribed spacer or the mature 5 S rRNA. The results further indicate the RNA is processed by an exonuclease activity which is limited primarily or entirely by helix I, the secondary structure formed between the mature and interacting termini. The 5 S RNA binding protein (YL3) also appears not to influence directly the maturation process, but rather to play a role in protecting the rRNA from further degradation by "housekeeping" nucleases. Taken together, the results continue to support a "quality control" function which helps to ensure that during maturation only normal precursors are processed and assembled into active ribosomes.
We have studied mechanisms by which leptin overexpression, which reduces body weight via anorexic and thermogenic actions, induces triglyceride depletion in adipocytes and nonadipocytes. Here we show that leptin alters in pancreatic islets the mRNA of the genes encoding enzymes of free fatty acid metabolism and uncoupling protein-2 (UCP-2). In animals infused with a recombinant adenovirus containing the leptin cDNA, the levels of mRNAs encoding enzymes of mitochondrial and peroxisomal oxidation rose 2- to 3-fold, whereas mRNA encoding an enzyme of esterification declined in islets from hyperleptinemic rats. Islet UCP-2 mRNA rose 6-fold. All in vivo changes occurred in vitro in normal islets cultured with recombinant leptin, indicating direct extraneural effects. Leptin overexpression increased UCP-2 mRNA by more than 10-fold in epididymal, retroperitoneal, and subcutaneous fat tissue of normal, but not of leptin-receptor-defective obese rats. By directly regulating the expression of enzymes of free fatty acid metabolism and of UCP-2, leptin controls intracellular triglyceride content of certain nonadipocytes, as well as adipocytes.
The azapirones, which are partial agonists of the serotonin (5-HT)1A receptor, possess anxiolytic activity. These agents may act at the pre- or postsynaptic 5-HT1A receptors, and involve the noradrenergic system. To determine whether these drugs activate noradrenergic neurons via 5-HT1A receptors, we have evaluated the expression of the immediate early gene c-fos in the locus coeruleus. Tandospirone and ipsapirone each induced expression of Fos protein in the noradrenergic neurons of the locus coeruleus of conscious rats. This effect was reversed by pretreatment with (+)-WAY100135, a specific 5-HT1A antagonist. These results clearly demonstrate that azapirones activate noradrenergic neurons via 5-HT1A receptors.
The 3' flanking region of the Escherichia coli rnpB gene-encoding M1 RNA, the RNA component of RNase P, contains a 113 bp repeated sequence. This sequence, successively reiterating 3.5 times, includes the region for intrinsic termination. In vivo termination of transcription occurs mostly at the first terminator (T1). Analysis of deletions at the 3' flanking region revealed that the second terminator (T2) and third (T3) are functional in vivo and that the sequences preceding the region coding for an RNA-terminator hairpin and U-rich 3' tail are essential for efficient termination. Transcripts terminating at T2 and T3 were also processed at the 3' end in a manner similar to those terminating at T1.
Leptin is currently believed to control body composition largely, if not entirely, via hypothalamic receptors that regulate food intake and thermogenesis. Here we demonstrate direct extraneural effects of leptin to deplete fat content of both adipocytes and nonadipocytes to levels far below those of pairfed controls. In cultured pancreatic islets, leptin lowered triglyceride (TG) content by preventing TG formation from free fatty acids (FFA) and by increasing FFA oxidation. In vivo hyperleptinemia, induced in normal rats by adenovirus gene transfer, depleted TG content in liver, skeletal muscle, and pancreas without increasing plasma FFA or ketones, suggesting intracellular oxidation. In islets of obese Zucker Diabetic Fatty rats with leptin receptor mutations, leptin had no effect in vivo or in vitro. The TG content was approximately 20 times normal, and esterification capacity was increased 3- to 4-fold. Thus, in rats with normal leptin receptors but not in Zucker Diabetic Fatty rats, nonadipocytes and adipocytes esterify FFA, store them as TG, and later oxidize them intracellularly via an "indirect pathway" of intracellular fatty acid metabolism controlled by leptin. By maintaining insulin sensitivity and preventing islet lipotoxicity, this activity of leptin may prevent adipogenic diabetes.
The Escherichia coli rnpA gene encodes C5 protein, the protein component of RNase P. The rnpA49 mutation renders the C5 protein thermosensitive, which results in thermosensitivity of RNase P function. The chromosomal DNA region from Brevibacterium albidum that complements the rnpA49 mutation was analysed. The gene capable of complementing the growth defect of an rnpA49 mutant strain at nonpermissive temperature was identified as the gene for an arginine tRNA with anticodon CCG by a deletion analysis combined with complementation assays. Transcription of the arginine tRNA gene carried on a multicopy plasmid was correlated with the complementation of the rnpA49 mutation, indicating that the gene product is indeed responsible for complementation of the rnpA49 mutation.
The region for intrinsic transcription termination of the Escherichia coli rnpB gene coding for M1 RNA is repeated three times. Each region encodes an RNA-terminator hairpin and U-rich 3' tail. Most transcription is terminated at the first terminator (T1), but a complete termination requires the second terminator (T2) and the third (T3). A deletion experiment, where deletions were extended from T3 into T1, was carried out to determine parameters affecting in vivo transcription termination at T1. The deletion of T3, T2 or the downstream sequences up to near the termination site of T1 showed little or no significant effect on transcription termination at T1. When the base corresponding to the acute termination position of T1 was included in the deletion, termination efficiency slightly decreased. For T1 bearing the deletion of the region encoding the RNA-terminator hairpin and U-rich 3' tail, termination was completely abolished. When T1 contained only the region encoding the RNA-termination hairpin without the U-rich 3' tail, termination still occurred even though the efficiency was low. Characteristics of rnpB termination were discussed with these results.
A cDNA clone encoding the Na+, K(+)-Atpase gamma-subunit polypeptide was cloned from a human fetal liver cDNA library. The deduced amino acid sequence of the protein comprised 58 amino acids with a calculated molecular weight of 6400 Da, and showed about 86% homology when compared with those of the bovine, rat and mouse Na+, K(+)-ATPase gamma-subunits published elsewhere. Northern blot analysis showed that the cDNA hybridized to a 0.7 kb mRNA which was expressed in human kidney, pancreas and fetal liver.
The promoter of the rnpB gene (encoding the RNA component of Escherichia coli RNase P) shares a consensus discriminator sequence, located between the -10 hexamer sequence and the transcription start site, with other promoters whose activities are repressed upon stringent condition. Under stringent conditions induced by seryl-tRNA starvation the transcription of the rnpB gene was repressed in wild type E. coli but not in a relaxed strain carrying a relA- mutation. Site-directed mutagenesis was carried out to examine sequences of the rnpB promoter necessary for stringent control. The results indicate that the discriminator region is responsible for the transcription repression of the rnpB gene during the stringent response and that both the content and position of GC pairs in the region determine the strength of negative stringent signals.
Macrobrachium rosenbergii that had been exposed individually for 24 h to 0 (control), 2, 5, 10 mg/L nitrite-N (nitrite as nitrogen) at 4. 3 and 7.7 pH levels were examined for hemolymph nitrite-N, oxyhemocyanin, protein, acid-base balance, ion concentrations, and ammonia-N (ammonia as nitrogen) excretion. Hemolymph oxyhemocyanin, protein, pH, HCO3- , TCO2, osmolality, and ion concentrations were inversely related to ambient nitrite-N concentration and were lower at pH 4.3. However, hemolymph nitrite-N, PO2 and PCO2 levels, and ammonia-N excretion were directly related to ambient nitrite-N, and were higher at pH 4.3. Ambient nitrite-N and pH level interacted to cause changes in hemolymph nitrite-N, oxyhemocyanin, protein, PO2, and pH levels. It is concluded that for M. rosenbergii following nitrite exposure, the incorporated nitrite causes a decrease of pH and an increase of PO2 in the hemolymph where it reduces oxyhemocyanin level; disturbs nitrogen excretion, ion regulation, and respiratory gas exchange; and may lead to a decrease of oxygen-carrying capacity, which are affected more at low pH.
Meiotic pairing of the X and Y chromosomes in Drosophila melanogaster males is mediated by the rDNA repeats, which are present in two tandem clusters, one in the centric X heterochromatin and the other near the base of the short arm of the Y chromosome. Deletion of the X chromosomal rDNA cluster disrupts X-Y pairing and causes high frequences of X-Y nondisjunction. Pairing can be partly restored by insertions of cloned complete rRNA genes or by rDNA fragments that include the intergenic spacer (IGS) region. A 240 bp repeated sequence in the IGS was shown to be effective in promoting pairing when present at copy numbers above five. This study further defines the rDNA sequences involved in mediating pairing. Germline insertions of a P element construct containing most of the rDNA transcription unit but no promoter or IGS region were obtained. Two single-copy insertions and four two-copy insertions proved unable to stimulate X-Y disjunction when located on an rDNA-deficient X chromosome. In addition, three insertions of a P element construct consisting of the IGS and promoter regions of the rDNA were characterized molecularly. These three insertions had previously been shown to range in pairing ability from very weak to quite strong. Molecular analysis revealed that the three insertions also vary in copy number of the 240 bp IGS repeat and that these structural differences correlate with the differences in pairing ability. These data indicate that 240 bp repeats are considerably more effective than other regions of the rDNA in stimulating chromosome pairing.
The whole-body acoustic startle response is a short-latency reflex mediated by a relatively simple neural circuit in the lower brainstem and spinal cord. The amplitude of this reflex is markedly enhanced by moderate fear levels, and less effectively increased by higher fear levels. Extensive evidence indicates that the amygdala plays a key role in the potentiation of startle by moderate fear. More recent evidence suggests that the periaqueductal gray is involved in the loss of potentiated startle at higher levels of fear. The influence of both structures may be mediated by anatomical connections with the acoustic startle circuit, perhaps at the level of the nucleus reticularis pontis caudalis. The present chapter reviews these data.
DNA replication of human papillomavirus type 18 is dependent on viral proteins E1 and E2 and the subsequent interaction of these proteins with the viral origin of replication. Using a site-directed mutagenesis analysis, we examined the sequence requirement for the DNA replication of the human papillomavirus type 18. We showed that both the E1BS palindrome and E2BS are the major determinants of the HPV replication efficiency. In particular, abolishing E2 binding sites demonstrated that E2BS makes a significant contribution towards HPV-18 DNA replication. Each part of the 18-bp inverted repeat sequence of the E1BS motif showed a clear functional difference between two regions: nt 13-21 (3' half segment) is evidently more important for replication than nt 4-12 (5' half segment). Besides E1BS and E2BS, cis-acting elements such as the poly-A6 track, perhaps the YY1 binding site, and the TATA box sequence within the origin region exhibited some contributions to optimum replication. In addition, inserting an enhancer region to the minimum origin DNA derivatives increased replication approximately 2-fold compared with the wild type levels and showed some compensational effects on loss of the cis-element within the HPV-18 minimum origin, suggesting that an enhancer region is required for efficient replication of the papillomavirus origin. These results suggest that the formation of an E1-E2-ori complex is important for replication, and other sequences near the E1 and E2 binding sites assist E1-E2-ori-mediated DNA replication in vivo.
The complete nucleotide sequence of a 1,513 bp fragment of Mycobacterium bovis BCG containing the secY gene homolog and partial adk gene that encodes an adenylate kinase has been determined. The secY gene of BCG has an open reading frame of 441 amino acids with homology to the SecY protein family. Comparative analyses of the deduced amino acid sequence of additional partial ORF revealed strong similarity to the known adenylate kinases.
A cDNA clone encoding human NADH:ubiquinone oxidoreductase (complex I of mitochondrial respiratory chain) MLRQ subunit was isolated from human fetal liver cDNA library. The clone contained an open reading frame of 246 by which predicted a protein comprising 81 amino acids with a calculated molecular weight of 9,370 Da. The deduced amino acid sequence exhibited 95% homology (88% identity and 7% favored substitution) to that of bovine MLRQ subunit. Northern analysis revealed that the cDNA clone hybridized with a 0.7 kb mRNA species which was present in all tissues examined. The expression level of the 0.7 kb mRNA in heart, skeletal muscle, and brain was higher than in other organs. Human MLRQ cDNA could cross-hybridize with the genomic DNAs from various species.
We designed the present study to examine the cross-sectional relation between age-related lens opacities and vitamin C supplement use over a 10-12-y period before assessment of lens status in women without diagnosed cataract or diabetes. This design avoids biased measurement of nutrient intake that results when knowledge of lens opacities influences nutrition-related behavior or its reporting. The participants were 247 Boston-area women aged 56-71 y selected from the Nurses' Health Study cohort with oversampling of women with high or low vitamin C intakes. Lens opacities were graded with the Lens Opacification Classification System II. Use of vitamin C supplements for > or = 10 y (n = 26) was associated with a 77% lower prevalence of early lens opacities (odds ratio: 0.23; 95% CI: 0.09, 0.60) at any lens site and a 83% lower prevalence of moderate lens opacities (odds ratio: 0.17; 95% CI: 0.03, 0.85) at any lens site compared with women who did not use vitamin C supplements (n = 141) after adjustment for age and other potentially confounding variables. Women who consumed vitamin C supplements for < 10 y showed no evidence of a reduced prevalence of early opacities. These data, together with data from earlier experimental and epidemiologic studies, suggest that long-term consumption of vitamin C supplements may substantially reduce the development of age-related lens opacities.