碩士 / 國立彰化師範大學 / 生物學系 / 97 / CHH (crustacean hyperglycemic hormone) is a neurohormone produced and secreted in the X-organ/sinus gland complex (XO/SG) of the eyestalk ganglia in crustacean. CHHs process molecular polymorphism, multiple functions and express in various tissues. Six CHHs mRNA (CHH1、CHH1-L、t-CHH1、CHH2、CHH2-L and t-CHH2) were discovered in the crayfish Procambarus clarkii (Yasuda et al., 1994；Kupper, 2002；Wu, 2005；Kung, 2009). In this study, western blot analyses indicated L/D-phe3 CHH1 present in sinus gland；L-phe3 CHH1-L present in thoracic ganglia. To purify these peptides from neurosecretory tissues, tissue extracts (sinus gland: SG; thoracic ganglia: TG) were chromatographically separated, followed by detection of immunoreactivity by enzyme-linked immunosorbent assay and identification by electrospray ionization quadrupole-time-of-flight mass spectrometer. Results showed that L-phe3 CHH1, D-phe3 CHH1, CHH2, N-unblocked L-phe3 CHH1 and truncated L/D-phe3 CHHs (CHH1 1-60、CHH2 1-61、CHH1 1-65、CHH2 1-65) were present in SG (which is located in the eyestalk ganglia, EG), while only CHH1-L was detected in TG. With regard to CHH1 contents in sinus gland (3.1 ×106 ± 7.0 ×104 pmol/mg proteins), thoracic ganglia (3.1 ×106 ± 7.0 ×104 pmol/mg proteins) and hemocyte (316.1 ± 5.5 pmol/mg proteins), were detected by using sandwich ELISA. While using sandwich ELISA, there are something interesting that the different combinations of capture antibody and biotin-labeled antibody cause difference in immunoreactivity. It’s speculated that present data may relation with the tertiary structure of CHH. This evidence reveal the tertiary structure of CHH and CHH-L are similar, and express in various tissues, but differ significantly in the expression content. In future work that need for further investigation in the tertiary structures and the functions CHH family.
| Identifer | oai:union.ndltd.org:TW/097NCUE5112006 |
| Date | January 2009 |
| Creators | Hsin-Ju Wu, 吳欣儒 |
| Contributors | Chi-Ying Lee, 李奇英 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 97 |
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