Molecular Analysis of the Ionotropic Glutamate receptor Genes in Vertebrates
| Autor: | Chen, Po Yuan, 陳柏苑 |
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| Rok vydání: | 1996 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 84 Glutamate receptors constitute the excitatory neurotransmitter system in the central nervous system (CNS). Based on the pharmacological and electrophysiological data, the mammalian ionotropic receptors are classified into three subtypes: {SYMBOL 97 \f "Symbol"}-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA), kainate (KA), and N-methyl-D- aspartate (NMDA)-preferring receptors. Eight cDNA clones encoding proteins with high amino acid sequence identities to the four mammalian AMPA-preferring receptor subunits were identified from Oreochromis sp. To study the numbers of vertebrate ionotropic glutamate receptor genes, degenerated primers derived from the most conserved regions of glutamate receptors were designed and used to amplify avian, reptilian, amphibian, and chondrichthyes glutamate receptor cDNAs by reverse transcription-PCR (RT-PCR). With the exception of bony fish (Oreochromis sp. and Tetraodon fluviatilis), there were only four types of cDNA molecules encoding AMPA receptor like subunits found in the brains of representative vertebral animals. PCR primers were designed according to the conserved amino acid sequence of the rat NMDAR1 transmembrane region, then were used to amplify fish brain cDNA encoding the NMDA receptor subunit. The fNR1 cDNA encoding protein with high amino acid sequence identity to rat NMDAR1 was identified by screening Oreochromis sp. brain cDNA library with the RT-PCR product as a probe. RT-PCR with designed degenerated primers was also performed to study the transcripts of NMDAR2-homolog in the brain of Oreochromis sp. A total of 7 different cDNA molecules homologous to the rat NMDAR2 subfamily was idnetified. These results imply that gene duplication of AMPA receptor like subunits and NMDAR2 subunits occurred during the evolution in bony fish. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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