Fat body is the site of vitellogenin synthesis in the soft tick,Ornithodoros moubata |
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Authors: | Yasuo Chinzei Isao Yano |
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Institution: | (1) Department of Medical Zoology, School of Medicine, Mie University, 514 Tsu, Japan;(2) National Research Institute of Aquaculture, Nakatsuhama, Nansei-cho, 516-01 Mie, Japan |
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Abstract: | Summary Vitellogenin (Vg) synthesis in cultured tissues was analysed biochemically in a soft tick,Ornithodoros moubata. Nine tissue fractions dissected from reproductive females were incubated in vitro in a specially designed Ringer containing35S-methionine. The synthesis of total protein and Vg was assayed by the radioactivity incorporated into precipitates with trichloroacetic acid and antivitellin (Vn)-serum, respectively. Fat body was the most active tissue in Vg synthesis, which comprised 46% of the Vg synthesis by all tissues and 42% of total protein synthesis by fat body. Protein synthesized by the fat body and precipitated with anti-Vn-serum was shown by electrophoresis and fluorography, to consist of six radioactive polypeptides corresponding to the components of Vg. Vg synthesized in cultured fat body was first accumulated in the tissue and secreted into the medium during incubation. Some tissues other than fat body showed low Vg synthesis (in each, less than 12% of total protein synthesis) which, however, may be due to contamination by fat body cells as seen with the scanning electron microscope (SEM). SEM also showed that fat body cells in the active stage of Vg synthesis expanded about 10-fold in length. Immunohistochemical analysis showed a very strong reaction with anti-Vn-IgG in the cytoplasm of fat body from reproductive females. Fat body from unfed females and other tissues including midgut, did not show any specific fluorescence. A positive reaction was obtained with developing oocytes. These results indicate that the fat body is the only site of Vg synthesis in this tick.Abbreviations
Vg
vitellogenin
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Vn
vitellin
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SDS
sodium dodecyl sulfate
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PAGE
polyacrylamide gel electrophoresis
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SEM
scanning electron microscopy
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TCA
trichloroacetic acid |
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