KONO Michiko Graduate School of Agricul.and Life Sci.Assistant Prof., 大学院・農学生命科学研究所, 助手 (70092202)
KOBAYASHI Makito Graduate School of Agricul.and Life Sci.Associate Prof., 大学院・農学生命科学研究所, 助教授 (30183809)
SUZUKI Yuzuru Graduate School of Agricul.and Life Sci.Associate Prof., 大学院・農学生命科学研究所, 助教授 (40107412)
WATANABE Toshiki Ocean Research Institute Associate.Professor., 海洋研究所, 助教授 (00272526)
NAGASAWA Hiromichi Graduate School of Agricul.aud Life Sci, Professor, 大学院・農学生命科学研究所, 教授 (60134508)
|Budget Amount *help
¥31,100,000 (Direct Cost : ¥31,100,000)
Fiscal Year 1997 : ¥5,200,000 (Direct Cost : ¥5,200,000)
Fiscal Year 1996 : ¥6,300,000 (Direct Cost : ¥6,300,000)
Fiscal Year 1995 : ¥19,600,000 (Direct Cost : ¥19,600,000)
(1) Seven CHH family peptides were isolated from the sinus gland in the eyestalk of kuruma prawn, and their primary structures were determined. Six peptides (Pej-SGP- I, II, III, V, VI, VII) possessed hyperglycemic activity and gonad-inhibiting activity. The other peptide (Pej-SGP- IV) showed molt-inhibiting activity.
(2) C-terminal peptides were synthesized for Pej-SGP- I, II, III, IV, V, /VI, and specific antisera against these peptides were raised. Using these antisera, neurosecretory cells in the X-organ in the eyestalk were identified immunohistochemically. Pej-SGP- I immunopositive cells were located in the MTGX-l, MTGX-3, MEGX-l, MEGX-2 and MIGX-l. Pej-SGP- II immunopositive cells were located in the MTGX-l and MTGX-2, whereas Pej-SGP- III, IV and V VI were found only in the MTGX-l. Furthermore, immunoreactivity of Pej-SGP- III and V/VI producing cells was decreased following ovarian maturation, indicating that these peptides are gonad-inhibiting hormone.
(3) Pej-SGP- I, III and IV V cDNAs were cloned and their sequences were determined. Recombinant Pej-SGP- IV showed molt-inhibiting activity.
(4) Yolk protein was purified from mature ovary and its N-terminal amino acid sequence was determined. Yolk protein cDNA was cloned by RT-PCR, Using this cDNA probe, it was confirmed that vitellin is synthesized in the ovary.
(5) Chitinase was purified and its primary structure was determined. Chitinase activity was increased by ecdysone administration.