水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2014年
1期
142-149
,共8页
杨明宇%王厚鹏%朱作言%孙永华
楊明宇%王厚鵬%硃作言%孫永華
양명우%왕후붕%주작언%손영화
斑马鱼%原始生殖细胞%表达模式%ca15b
斑馬魚%原始生殖細胞%錶達模式%ca15b
반마어%원시생식세포%표체모식%ca15b
Zebrafish%Primordial germ cells%Expression pattern%ca15b
高保真PCR克隆获得了ca15b基因的全长,利用胚胎整体原位杂交等技术研究了ca15b基因在斑马鱼早期发育过程中的动态表达。结果发现, ca15b在斑马鱼早期发育过程中存在显著的原始生殖细胞(Primordial germ cell, PGC)特异表达模式。ca15b是一个母源性表达的基因：在分裂期的胚胎中,其mRNA集中分布于位于分裂沟的生殖质(Germ plasm)；从囊胚期开始,可以观察到其在PGC中的特异表达；在原肠胚中,其mRNA在体细胞中急剧降解,仅特异表达于PGC,这一表达特征一直持续到受精后1d的胚胎。将体外合成的包含5′UTR和3′UTR的ca15b全长mRNA注射到斑马鱼胚胎后,仅能增强原肠期之前胚胎中ca15b的整体杂交信号；在原肠胚期之后,注射的 mRNA在体细胞中快速降解。这提示在 ca15b mRNA上可能存在某种转录后调控其在早期胚胎体细胞中降解而在PGC中稳定存在的机制。
高保真PCR剋隆穫得瞭ca15b基因的全長,利用胚胎整體原位雜交等技術研究瞭ca15b基因在斑馬魚早期髮育過程中的動態錶達。結果髮現, ca15b在斑馬魚早期髮育過程中存在顯著的原始生殖細胞(Primordial germ cell, PGC)特異錶達模式。ca15b是一箇母源性錶達的基因：在分裂期的胚胎中,其mRNA集中分佈于位于分裂溝的生殖質(Germ plasm)；從囊胚期開始,可以觀察到其在PGC中的特異錶達；在原腸胚中,其mRNA在體細胞中急劇降解,僅特異錶達于PGC,這一錶達特徵一直持續到受精後1d的胚胎。將體外閤成的包含5′UTR和3′UTR的ca15b全長mRNA註射到斑馬魚胚胎後,僅能增彊原腸期之前胚胎中ca15b的整體雜交信號；在原腸胚期之後,註射的 mRNA在體細胞中快速降解。這提示在 ca15b mRNA上可能存在某種轉錄後調控其在早期胚胎體細胞中降解而在PGC中穩定存在的機製。
고보진PCR극륭획득료ca15b기인적전장,이용배태정체원위잡교등기술연구료ca15b기인재반마어조기발육과정중적동태표체。결과발현, ca15b재반마어조기발육과정중존재현저적원시생식세포(Primordial germ cell, PGC)특이표체모식。ca15b시일개모원성표체적기인：재분렬기적배태중,기mRNA집중분포우위우분렬구적생식질(Germ plasm)；종낭배기개시,가이관찰도기재PGC중적특이표체；재원장배중,기mRNA재체세포중급극강해,부특이표체우PGC,저일표체특정일직지속도수정후1d적배태。장체외합성적포함5′UTR화3′UTR적ca15b전장mRNA주사도반마어배태후,부능증강원장기지전배태중ca15b적정체잡교신호；재원장배기지후,주사적 mRNA재체세포중쾌속강해。저제시재 ca15b mRNA상가능존재모충전록후조공기재조기배태체세포중강해이재PGC중은정존재적궤제。
Primordial germ cells (PGCs) are the ancestor cells of adult gametes at embryonic stage. PGCs-specific gene expression plays a fundamental role in germ cell induction, development, migration and differentiation. Understanding the functional role of PGCs-specifically expressed genes could provide us some tools to perform gene and embryo ma-nipulations. Screening a public database of zebrafish (http://www.zfin.org) we found a PGCs-specific candidate gene, carbonic anhydrase 15b (ca15b). To confirm this, whole-mountin situ hybridization and other technologies were uti-lized to analyze the expression ofca15bduring zebrafish early development. We observed thatca15b is a maternally expressed gene and it is highly expressed in germ plasm during cleavage stage. The expression ofca15b at mRNA level was specifically detected in the PGCs at blastula stage. At gastrula stage,ca15b is still strictly expressed in PGCs while its expression is undetectable in somatic cells. This specific expression pattern ofca15b persisted until one day post-fertilization. More importantly, the post-transcriptional regulation elements that mediate PGCs-specific expression ofca15b may locate in its coding sequence.
