Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea
碩士 === 國立彰化師範大學 === 生物學系 === 93 === Crustacean hyperglycemic hormone (CHH) is a neuropeptide original identified from the X-organ (XO)-sinus gland (SG) complex in the crustacean eyestalk. In the present work, three full-length CHH cDNAs were cloned from the mud crab Scylla olivacea eyestalk ganglia...
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ndltd-TW-093NCUE51120162016-06-03T04:13:56Z http://ndltd.ncl.edu.tw/handle/13309963145382498785 Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea 紅腳蟳(Scyllaolivacea)甲殼類升血糖荷爾蒙異構型之分子特徵與生理功能之研究 Kuo-Wei Tsai 蔡國瑋 碩士 國立彰化師範大學 生物學系 93 Crustacean hyperglycemic hormone (CHH) is a neuropeptide original identified from the X-organ (XO)-sinus gland (SG) complex in the crustacean eyestalk. In the present work, three full-length CHH cDNAs were cloned from the mud crab Scylla olivacea eyestalk ganglia (ES), thoracic ganglia (TG), and per- icardial organ (PO) by RT-PCR and rapid amplification of cDNA end (RACE). PO-CHH and TG-CHH have the same deduced amino acid sequence. TG/PO-CHH and sinus gland CHH (SG-CHH) share an identical N-terminal sequence (amino acids 1-40), but the remaining sequences (amino acids 41-73 or 41-75), differ considerably. The sequence evidence suggests that these two isoforms are derived a Chh gene transcribed in an alternative splicing manner. With the use of a two-step HPLC purification procedure and a CHH isoform- specific ELISA, the two CHH isoforms were isolated from sinus gland and pericardial organ, respectively. Mass spectrometric analysis indicates that the molecular masses for the native SG-CHH and TG/PO-CHH are 8379 Da and 8478 Da, respectively, which are consistent with the estimated values based on the deduced sequences. It also suggests that the two CHHs have a blocked N- terminus, and the C-terminus of SG-CHH is amidated, but TG/PO-CHH has a free C-terminus. The partial amino acid sequences of SG-CHH and TG/PO- CHH were determined by MALDI MS/MS analysis of trypsin-digested peptide fragments. The results show that the amino acid sequences of these peptide fragments were identical to the deduced amino acid sequences of the SG-CHH and TG/PO-CHH cDNAs. In addition, SG-CHH (10 pmole) exhibited a hyper- glycemic activity in vivo, whereas the same dosage of TG/PO-CHH did not elicit a hyperglycemic response. The physiological function of TG/PO-CHH remains to be clarified. We speculate that TG/PO-CHH may have regulatory functions differ from those of SG-CHH. Chi-Ying Lee 李奇英 2005 學位論文 ; thesis 114 zh-TW |
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碩士 === 國立彰化師範大學 === 生物學系 === 93 === Crustacean hyperglycemic hormone (CHH) is a neuropeptide original identified from the X-organ (XO)-sinus gland (SG) complex in the crustacean eyestalk. In the present work, three full-length CHH cDNAs were cloned from the mud crab Scylla olivacea eyestalk ganglia (ES), thoracic ganglia (TG), and per- icardial organ (PO) by RT-PCR and rapid amplification of cDNA end (RACE). PO-CHH and TG-CHH have the same deduced amino acid sequence. TG/PO-CHH and sinus gland CHH (SG-CHH) share an identical N-terminal sequence (amino acids 1-40), but the remaining sequences (amino acids 41-73 or 41-75), differ considerably. The sequence evidence suggests that these two isoforms are derived a Chh gene transcribed in an alternative splicing manner. With the use of a two-step HPLC purification procedure and a CHH isoform- specific ELISA, the two CHH isoforms were isolated from sinus gland and pericardial organ, respectively. Mass spectrometric analysis indicates that the molecular masses for the native SG-CHH and TG/PO-CHH are 8379 Da and 8478 Da, respectively, which are consistent with the estimated values based on the deduced sequences. It also suggests that the two CHHs have a blocked N- terminus, and the C-terminus of SG-CHH is amidated, but TG/PO-CHH has a free C-terminus. The partial amino acid sequences of SG-CHH and TG/PO- CHH were determined by MALDI MS/MS analysis of trypsin-digested peptide fragments. The results show that the amino acid sequences of these peptide fragments were identical to the deduced amino acid sequences of the SG-CHH and TG/PO-CHH cDNAs. In addition, SG-CHH (10 pmole) exhibited a hyper- glycemic activity in vivo, whereas the same dosage of TG/PO-CHH did not elicit a hyperglycemic response. The physiological function of TG/PO-CHH remains to be clarified. We speculate that TG/PO-CHH may have regulatory functions differ from those of SG-CHH.
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author2 |
Chi-Ying Lee |
author_facet |
Chi-Ying Lee Kuo-Wei Tsai 蔡國瑋 |
author |
Kuo-Wei Tsai 蔡國瑋 |
spellingShingle |
Kuo-Wei Tsai 蔡國瑋 Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
author_sort |
Kuo-Wei Tsai |
title |
Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
title_short |
Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
title_full |
Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
title_fullStr |
Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
title_full_unstemmed |
Comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab Scylla olivacea |
title_sort |
comparative studies on molecular characterizations and physiological functions of crustacean hyperglycemic hormone isoforms in the mud crab scylla olivacea |
publishDate |
2005 |
url |
http://ndltd.ncl.edu.tw/handle/13309963145382498785 |
work_keys_str_mv |
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