碩士 / 國立彰化師範大學 / 生物學系 / 101 / Crustacean hyperglycemic hormone (CHH) was originally identified in a neuroendocrine system－the X-organ/sinus gland complex. In this study, a cDNA(Prc-CHH) encoding CHH precursor was cloned from the hemocyte and hematopoietic cells (HpT cells) of the crayfish Procambarus clarkii. Analysis of tissues by a CHH-specific enzyme-linked immunosorbent assay (ELISA)confirmed the presence of CHH in hemocytes and HpT, the levels of which were much lower than those in the sinus gland, but much higher than those in the
thoracic and cerebral ganglia. Total hemocytes were separated by density gradient centrifugation into layers of hyaline cells (HCs), semi-granular cells 1-3(SGCs 1-3), and granular cells (GCs). Analysis of extracts of each layer using ELISA revealed that CHH is present in GCs (202.8 ± 86.7 fmol/mg protein) and SGCs 1-3 (204.424 ± 27.656, 235.768 ± 37.226, 497.8 ± 49.4 fmol/mg protein), but not in HCs. In addition, immunohistochemical analysis demonstrated the presence of CHH immunoreactivity in granules of hemocyte and HpT. In addition, immunohistochemical stain analysis demonstrated the presence of CHH immunoreactivity in granules of SGCs, GCs, and HpT. After white spot syndrome virus (WSSV) infection, CHH gene expression levels in HpT were significantly increased at 12, 24, and 48 hr post infection (hpi), and in hemocyte significantly increased at 48 hpi. CHH peptide levels were not significantly changed after viral infection at any time points measured in either hemocytes or HpT. These data for the first time confirm that a crustacean neuropeptide-encoding gene is expressed in immune-related tissues and its expression in hemocytes is cell type-specific. The CHH in immune-related tissue is sensitive response to pathogen-related stresses.
| Identifer | oai:union.ndltd.org:TW/101NCUE5112114 |
| Date | January 2013 |
| Creators | Tsai,Wei-Shiun, 蔡偉勳 |
| Contributors | Lee Chi-Ying, 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 | 62 |
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