IDENTIFICATION AND EXPRESSION OF BOULE, DAZL AND VASA IN GERM CELLS OF THE ASIAN SEABASS

Abstract

THE ASIAN SEABASS (LATES CALCARIFER) IS A HERMAPHRODITIC FISH, WHICH UNDERGOES SEQUENTIAL MALE TO FEMALE SEX REVERSAL WHICH OFFERS AN EXCELLENT SYSTEM TO STUDY THE ROLES OF SOMATIC CELLS AND GERM CELLS IN THE CONTEXT OF BOTH SEX DIFFERENTIATION AND SEX REVERSAL. THIS THESIS WORK WAS AIMED AT THE IDENTIFICATION AND CHARACTERIZATION OF MULTIPLE GERMLINE GENES FOR ANALYZING STAGE-SPECIFIC EXPRESSION DURING GAMETOGENESIS IN THE ASIAN SEABASS. GENES BOULE, DAZL AND VASA, WHICH ENCODE HIGHLY CONSERVED RNA BINDING PROTEINS, WERE IDENTIFIED BASED ON CLONING AND SEQUENCE ANALYSES. CHROMOGENIC IN SITU HYBRIDIZATION REVEALED THEIR GERM CELL-SPECIFIC EXPRESSION IN BOTH SEXES. TWO-COLOR FLUORESCENCE IN SITU HYBRIDIZATION (FISH) REVEALED DIFFERENTIAL TEMPOROSPATIAL PATTERNS OF VASA, DAZL AND BOULE RNA EXPRESSION TOGETHER WITH NUCLEAR STAINING, MORPHOLOGY AND TISSUE ARCHITECTURE AND ALLOWED FOR STAGING DEVELOPING GERM CELLS. FURTHERMORE, A THREE-COLOR FISH PROCEDURE WAS DEVELOPED THAT ALLOWS FOR SIMU