由於目前石斑魚生殖細胞早期發育及性腺形成分子機制相關研究甚少，故本研究希望以選殖生殖細胞追蹤標誌Vasa基因探討石斑魚原始生殖細胞發生、遷移過程及生殖腺早期發育。此外也探討生殖腺發育相關基因HSC70 及SPARC對於石斑魚生殖腺發育所扮演之角色。所選殖龍膽石斑魚Vasa 基因全長為2,399 bp，蛋白質轉譯編碼區為1,929 bp，轉譯出643個胺基酸，分子量約為69.62 kDa。龍膽石斑魚Vasa蛋白質序列具8個DEAD-box蛋白質家族保守性序列。由親緣演化樹及蛋白質序列相似度分析，其分屬於海水硬骨魚類且與其它魚類具高度相似度（73-85%）。龍膽石斑魚Vasa特異表現在生殖腺且基因表現量隨著生殖腺發育增加。以化學組織免疫染色觀察Vasa專一性表現在生殖細胞中。由全魚原位雜交及免疫螢光染色發現受精後第三天原始生殖細胞已移動至生殖脊區。此外，由結果可知HSC70於生殖細胞開始大量發育時表現量增加，而SPARC大量表現於管腔開始發育形成之三月齡石斑魚。由此推論HSC70可能參與生殖細胞生成過程中，SPARC與早期生殖管腔形成與發育相關。

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.

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