In vitro growth of corpora allata fromDiploptera punctata |
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Authors: | W. -H. Tsai G. L. Holbrook C. Schal A. -S. Chiang |
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Affiliation: | (1) Institute of Life Science, National Tsing-Hua University, Hsinchu, Taiwan 30043, Republic of China;(2) Department of Entomology, North Carolina State University, Box 7613, 27695-7613 Raleigh, North Carolina |
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Abstract: | Summary An in vitro organ culture system was established to support growth of corpora allata from the cockroachDiploptera punctata. During a 1-wk incubation in L-15B medium supplemented with 10% fetal bovine serum (FBS) and 10% cockroach hemolymph, adult male corpora allata exhibited a cycle of de novo DNA synthesis followed by cell division. The number of S-phase cells and metaphase cells per corpus allatum were counted from whole-mount monolayers after labeling in vitro with 5′-bromo-2′-deoxyuridine and exposure to colchicine, respectively. While both FBS and cockroach hemolymph were essential for proliferation of allatal cells, the growth-promoting effect of insect hemolymph was not species-specific and adult female hemolymph was more potent than hemolymph from adult males. Furthermore, DNA synthesis of corpus allatum cells was stimulated in vitro by 20-hydroxyecdysone. This sensitive assay system will be of immense utility in the search for allatal growth factors. |
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Keywords: | corpora allata juvenile hormone cockroach insect organ culture DNA synthesis Diploptera punctata |
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