title: Hatching mechanism of the Chinese soft-shelled turtle Pelodiscus sinensis.
abstract: The mechanism by which the embryo hatches out of the egg envelope, the vitelline membrane and egg white, was studied in the Chinese soft-shelled turtle Pelodiscus sinensis. The cDNA of the turtle hatching enzyme (HE) was 1555bp-long and a mature enzyme of 321 amino acids. The mature HE was composed of an astacin protease domain of 200 amino acids and a CUB domain of 121 amino acids, and the estimated molecular size was 35,311. The protease domain contained two active site consensus sequences, HExxHxxGFxHExxRxDR and MHY. An immunoblotting test of an extract of allanto-chorions revealed a 40-kDa band by cross-reaction with the anti-Xenopus HE antiserum. The first change in the envelopes was the appearance of a hole, 1mm in diameter, at the location around the animal pole of day 8 incubation eggs. A cluster of tall cells, forming a circle in the avascular chorion of day 8 embryos and facing the edge of the hole, had various sizes of inclusion bodies and secretory granules that were labeled by immuno-electron microscopic staining with the antiserum. The egg envelopes were degraded gradually from the animal pole side towards the vegetal pole side in accordance with translocation of the avascular site of the chorion in the same direction. Labeled cells degenerated, presumably when the chorion was underlain by allantois in succeeding developmental stages. The vitelline membrane and egg white were totally digested, presumably by secreted HE, during the hatching period and were consumed for embryonic growth.
authors.affiliation: Department of Materials and Life Sciences, Sophia University, Tokyo, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Yasumasu
authors.forename: Moeko
authors.initials: M
authors.lastname: Uzawa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Iwasawa
authors.forename: Norio
authors.initials: N
authors.lastname: Yoshizaki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D014426:Turtles / Q000196:embryology* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D008666:Metalloendopeptidases; C024344:envelysin