Molecular Cloning of Crustacean Biogenic Amine Receptors

• Main Authors: Shyh-Chi Chen, 陳仕錤
• Other Authors: Chi-Ying Lee
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/82841277171769358893

碩士 === 國立彰化師範大學 === 生物學系 === 90 === Biogenic amines, functioning as eurotransmitters/neurohormones, are involved in a wide variety of physiological processes. In crustaceans, several biogenic amines, including serotonin (5-HT), dopamine, and octopamine have been found to regulate the release of several peptide hormones from a neuroendocrine tissues, X-organ-sinus gland complex, located within the eyestalks of decapods. In order to advance our knowledge regarding neuronal regulation of hormonal secretion in crustaceans, the objective of this study was to molecularly clone biogenic amine receptors from the eyestalk ganglia, where the X-organ-sinus gland receives neuronal signals that subsequently regulate the secretion of neuropeptides. Primers were designed based on the conserved sequences located in the 2nd, 6th, 7th transmembrane domains of the biogenic amine receptors and used for PCR amplification of eyestalk ganglia cDNA. PCR products of expected size were purified, cloned, and sequenced. Sequence analysis revealed that they contain sequence characteristics of biogenic amine receptor genes, including octopamine/tyramine, dopamine, serotonin, and histamine receptors. Primers were further designed based on some of the amplified sequences and used for RACE amplification of 3’- and 5’-ends. However, sequence analysis of these RACE PCR products indicated that they share high similarity with olfactory receptors, but not biogenic amine receptors. Currently, the amplified sequences showing characteristics of biogenic amine receptors are used as probes in screening a cDNA library in order to obtain full-length sequences of novel crustacean biogenic amine receptor genes.