碩士 / 國立彰化師範大學 / 生物學系 / 104 / Crustacean hyperglycemic hormone (CHH) is the hormone which is produced and secreted by eyestalk ganglion of crustaceans. The most important physiological function of CHH is influencing the glycogen metabolism in the target tissues (muscle, liver, pancreas and other tissues) and promoting the glucose mobilization. It may affect crustacean immune function indirectly. The previous study showed that the CHH could promote the glycolysis and the pentose phosphate pathway which might affect the crustacean immune functions indirectly. On the other hand, viruses rely on their host cells to the supply the biosynthetic precursors and energy which were necessary for their replication. The viruses might alter the host cells metabolic process in order to satisfy their demand after a successful infection.
Previous data show that white spot syndrome virus (WSSV) induced the crustacean host to release crustacean glycemic hormone fast and persistently. However, the blood glucose of host was not increasing significantly in the hemolymph. Therefore, CHH-induced glycemic response was likely to be offset by CHH-induced glycolysis response, or virus applied synthetic carbohydrate resources on replication.
In this study, by using in vitro transcription method to product CHH double-stranded RNA. We established the application of RNAi technology on crayfish (procambarus carkii), to observe the changes of physiological and metabolic in the shrimp after CHH gene silence.
The experiment animals received saline or CHH dsRNA in saline (38 ug / g crayfish). Hemolymph and anatomy organizations (eyestalk ganglia, hematopoietic tissue, muscle, liver and pancreas) were collected at just before the infection treatment (0 hours) and the time points after injection (24, 48 hours) for subsequent analysis.
The results showed that the CHH gene expression was significantly decreased in eyestalk ganglion in 48 hours after CHH dsRNA injection. In addition, the metabolites of the muscle and hepatopancreas were analyzed via the nuclear magnetic resonance spectroscopy (NMR). The principal of component analysis (PCA) showed that the major metabolite signals in muscle were significantly divided into different quadrant zones after CHH gene silence treatment, compare the CHH dsRNA injection group with the untreated group and the saline injection group. Moreover, the analysis of the compounds which is significantly influenced was performed to understand the biochemical pathways. The present results showed that the CHH was involved not only in regulating the carbohydrate metabolism, but also in the biochemical pathways of amino acids and immunity.
| Identifer | oai:union.ndltd.org:TW/104NCUE5112003 |
| Date | January 2016 |
| Creators | 蔡士甫 |
| Contributors | 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 | 67 |
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