猪GDF11基因部分序列的克隆及组织表达分析

Cloning of partial porcine GDF11 and its mRNA expression in tissues

为了探讨生长分化因子11(Growth differentiation factor11,GDF11,又名BMP11)在胸腰椎数变异中的作用,本试验克隆了该基因包含外显子2在内的部分编码区,并进一步采用RT-PCR技术对其在猪胚胎和初生仔猪中的表达进行了分析。结果表明,在35d的猪胚胎中,后肢、牙龈、脑、肝脏、肾脏、胸椎、腰椎各组织均有明显的表达,而在前肢、眼、心脏、肺脏中的表达较弱,在颈椎和荐尾椎中没有观察到GDF11的表达。在45d猪胚胎的后肢、脑、眼、胸椎组织中GDF11的表达较强,而在前肢、牙龈、肺脏、肾脏、腰椎和荐尾椎的表达相对较弱,在肝脏中的表达极其微弱。在心脏和颈椎中没有检测到GDF11的表达。在55d的猪胚胎中,前肢、后肢、脑、眼、肝脏、颈椎、胸椎、腰椎组织中有明显的表达,肺脏和肾脏组织中的表达较强,牙龈和荐尾椎中的表达较弱,而在心脏中没有检测到GDF11的表达。3d仔猪的后肢、牙龈、脑、肾脏和腰椎组织中GDF11有明显的表达,脾脏组织的表达量较高,前肢、肝脏、心脏、背腰最长肌和肺脏中的表达相对较弱,在眼、颈椎和荐尾椎中的表达极弱,在胸椎中没有检测到表达。在所检测的不同时期的所有组织中,脑和肾脏组织表达明显地高于其他组织。

Growth differentiation factor 11,also known as BMP11,is a secreted protein belonging to the TGF-β gene superfamily and BMPs subfamily.It is involved in the induction of dorsal mesoderm and plays important roles in the formation and development of vertebrae,kidney and neural tissues.To investigate the role of GDF11 in the number variation of thoracic and lumbar vertebra,the present study first cloned the partial region including exon 2 of porcine GDF11,then analyzed its mRNA expression in porcine embryos and newborn piglets.The results are as follows：In 35-day-embryos,high GDF11 expression was detected in hindlimb,gingiva,brain,liver,kidney,thoracic vertebra and lumbar vertebra,weak expression in forelimb,eye,heart and lung,while no expression was detected in cervical vertebra and caudal vertebra.In 45-day-embryos,GDF11 mRNA expression is high in hindlimb,brain,eye and lumbar vertebra;low in the forelimb,gingiva,lung,kidney,lumbar vertebra and caudal vertebra;while not detected in the heart and cervical vertebra.In 55-day-embryo,GDF11 mRNA expression is high in forelimb,hindlimb,brain,eye,liver,cervical vertebra,thoracic vertebra and lumbar vertebra,less in lung and kidney tissues,weak in gingiva and caudal vertebra.No expression was detected in heart.In 3-day-piglets,GDF11 mRNA expression is high in hindlimb,gingiva,brain,kidney,lumbar vetebra and spleen,low in forelimb,liver,heart,longissimus dorsi muscle and lung,exremely low expression in the eye,cervical vertebra and caudal vertebra.No expression was detected in thoracic vertebra.The GDF11 mRNA expression is high in brain and kidney in all of the embryos and piglets investigated.The GDF11 expression study is a basis for further revealing a relationship between GDF11 expression and the number of thoracic vertebra and lumbar vetebra.