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Used Polymerase Chain Reaction (used + polymerase_chain_reaction)
Selected AbstractsAssociation of vasopressin 1a receptor levels with a regulatory microsatellite and behaviorGENES, BRAIN AND BEHAVIOR, Issue 5 2005E. A. D. Hammock Vasopressin regulates complex behaviors such as anxiety, parenting, social engagement and attachment and aggression in a species-specific manner. The capacity of vasopressin to modulate these behaviors is thought to depend on the species-specific distribution patterns of vasopressin 1a receptors (V1aRs) in the brain. There is considerable individual variation in the pattern of V1aR binding in the brains of the prairie vole species, Microtus ochrogaster. We hypothesize that this individual variability in V1aR expression levels is associated with individual variation in a polymorphic microsatellite in the 5, regulatory region of the prairie vole v1ar gene. Additionally, we hypothesize that individual variation in V1aR expression contributes to individual variation in vasopressin-dependent behaviors. To test these hypotheses, we first screened 20 adult male prairie voles for behavioral variation using tests that measure anxiety-related and social behaviors. We then assessed the brains of those animals for V1aR variability with receptor autoradiography and used polymerase chain reaction to genotype the same animals for the length of their 5, microsatellite polymorphism in the v1ar gene. In this report, we describe the results of this discovery-based experimental approach to identify potential gene, brain and behavior interrelationships. The analysis reveals that V1aR levels, in some but not all brain regions, are associated with microsatellite length and that V1aR levels in those and other brain regions correlate with anxiety-related and social behaviors. These results generate novel hypotheses regarding neural control of anxiety-related and social behaviors and yield insight into potential mechanisms by which non-coding gene polymorphisms may influence behavioral traits. [source] Immune activation upregulates lysozyme gene expression in Aedesaegypti mosquito cell cultureINSECT MOLECULAR BIOLOGY, Issue 6 2000Y. Gao Abstract After stimulation with heat-killed bacteria, cultured cells from the mosquito Aedesaegypti (Aag-2 cells) secreted an induced protein with a mass of , 16 kDa that cross-reacted with antibody to chicken egg lysozyme. To investigate whether lysozyme messenger RNA is induced in bacteria-treated cells, we used polymerase chain reaction-based approaches to obtain the complete lysozyme cDNA from Aag-2 cells. The deduced protein contained 148 amino acids, including a 23 amino acid signal sequence. The calculated mass of the precursor protein is 16 965 Da, which is processed to yield a mature lysozyme of 14 471 Da with a calculated pI of 10.1. The lysozyme from Ae. aegypti shared 50% amino acid identity with lysozymes from Anophelesgambiae and Anophelesdarlingi, which in turn shared 70% identity between each other. Northern analysis with the lysozyme cDNA probe showed induction of a 1.3 kb messenger RNA during the first 3 h after treatment of Aag-2 cells with heat-killed bacteria, followed by maximal expression 12,36 h after treatment. Southern analysis suggested that the gene likely occurs as a single copy in the genome of Aag-2 cells. [source] Molecular analysis of thiopurine S -methyltransferase alleles in Taiwan aborigines and TaiwaneseJOURNAL OF CLINICAL PHARMACY & THERAPEUTICS, Issue 1 2006H-F. Lu MS Summary Background:, Thiopurine S -methyltransferase (TPMT) is a cytosolic enzyme involved in the metabolism of these thiopurine drugs. Methylation of thiopurine drugs by TPMT competes with the formation of their active 6-thioguanine nucleotide metabolite, thereby potentially modulating the therapeutic and toxic effects of these drugs. Objective:, To analyze the thiopurine S -methyltransferase allelic frequencies in Taiwan aborigines and Taiwanese. Methods:, We used polymerase chain reaction,restriction fragment length polymorphism method to determine the allelic frequencies of TPMT variants (TPMT*1,TPMT*8) in 409 Taiwan aborigines and 117 Taiwanese. Results and discussion:, The results showed that the allelic frequencies of TPMT*1 were 99·88% and 98·72% for Taiwan aborigines and Taiwanese respectively. The allelic frequencies of TPMT*3C were 0·12% and 1·28% for Taiwan aborigines and Taiwanese respectively. No TPMT*2, 3A, 3B, 3D and 4,8 were found in these populations. Conclusion:, Our results provide useful information for using thiopurine drugs in these populations. [source] Developmental and adult expression of semaphorin 2a in the cricket Gryllus bimaculatus,THE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 1 2007Kristen R. Maynard Abstract Developmental guidance cues act to direct growth cones to their correct targets in the nervous system. Recent experiments also demonstrate that developmental cues are expressed in the adult mammalian nervous system, although their function in the brain is not yet clear. The semaphorin gene family has been implicated in the growth of dendrites and axons in a number of different species. While the expression of semaphorin and its influence on tibial pioneer neurons in the developing limb bud have been well characterized in the grasshopper, the expression of semaphorin 2a (sema2a) has not been explored in the adult insect. In this study we used polymerase chain reaction (PCR) with degenerate and gene-specific primers to clone part of the secreted form of sema2a from Gryllus bimaculatus. Using in situ hybridization and immunohistochemistry, we confirmed that sema2a mRNA and protein expression patterns in the embryonic cricket were similar to that seen in the grasshopper. We also showed that tibial neuron development in crickets was comparable to that described in grasshopper. An examination of both developing and adult cricket brains showed that sema2a mRNA and protein were expressed in the Kenyon cells in mushroom bodies, an area involved in learning and memory. Sema2a expression was most obvious near the apex of the mushroom body in a region surrounding the neurogenic tip, which produces neurons throughout the life of the cricket. We discuss the role of neurogenesis in learning and memory and the potential involvement of semaphorin in this process. J. Comp. Neurol. 503:169,181, 2007. © 2007 Wiley-Liss, Inc. [source] | ||||||||||