Characterization of gonad formation process in protogynous grouper by using gonadal development and germ cell related tracing markers

Abstract

Studies about early germ cells development and gonad formation of grouper species were limited Therefore this study characterized the ontology of primordial germ cells (PGCs) and gonadogenesis by cloning the potential germ cell marker Vasa from the giant grouper Epinephelus lanceolatus The full length of Vasa contained 1 929 bp of open reading frame which generated 643 amino acid protein with the eight conserved domains belong to DEAD box family Phylogenetic tree analysis and comparisons of the deduced amino acid sequence with other vertebrates revealed the high homology (73-85%) and belonged to marine teleost Moreover giant grouper Vasa gene was highly expressed in gonad and increased with gonadal development IHC stain showed ggVasa distributed in the cytoplasm of germ cells Whole-mount in situ hybridization and immunostaining data indicated PGCs migrated to germinal ridge at 3 dpf Besides the roles of HSC70 and SPARC were investigated as well The results of qRT-PCR and IHC stain showed HSC70 was significantly higher during germ cell proliferation Besides SPARC expression was higher and strongly detected in the reproductive tract of 3 month-old giant grouper Consequently HSC70 may be involved in gametogenesis and SPARC may play specific roles in reproductive tract formation

Date of Award	2014 Sept 10
Original language	English
Supervisor	Tzong-Yueh Chen (Supervisor)