| Summary: | 碩士 === 國立彰化師範大學 === 生物學系 === 102 === CHH (crustacean hyperglycemic hormone) was originally identified in a crustacean neurosecretory complex, the X-organ/sinus gland. Previous studies, CHH might directly or indirectly regulate carbohydrate metabolism, thereby affecting the immune system of crustaceans. Therefore, this study first established RNA interference on Procambarus clarkii, and confirmed CHH gene silencing by real-time quantitative polymerase chain reaction. It might cause physiological changes in metabolism and hemocyte numbers. The results showed that CHH double-stranded RNA (CHH dsRNA) (38 μg / g crayfish) –injected. At 72, 96 and 120 hours post injection, a significant effect CHH expression in the eyestalk ganglia, and CHH levels also significantly reduced in the sinus glands; In the hematopoietic tissue and hemocyte, the CHH gene expression and protein decreased at the 24 and 48 hours post injection, while changes in hemolymph CHH levels was similar. In physiological metabolism, analysis of glucose and lactate levels at 72, 96 and 120 hours post injection in the hemolymph, changes were decreased and increased, respectively. The number of granular cells, semi-granular cells and hyaline cells were significantly reduced at 24 hours post injection, and the percentage of granular cells and hyaline cell were increased by 10%, while the percentage of semi-granular cells was decreased by 20%. In addition, analysis of metabolism by nuclear magnetic resonance (NMR), initially found to effect on principal component analysis (PCA) by CHH gene silencing. The data showed, sample signals can be divided into two main components of the community, and analysis of biological pathway involved by changes of metabolism intermediates, including the synthesis and metabolism of carbohydrate, lipid, nucleotide and amino acid. Summary, at this study was the first time to establish RNA interference in P. clarkii, and confirmed the CHH gene silencing, but also a preliminary understanding of the metabolism by NMR analysis. Therefore, the CHH might be regulating physiological, and not limited to regulating carbohydrate metabolism, also contain such as synthesis of lipid and immunity.
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