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日本沼虾胰岛素样雄性腺激素结合蛋白基因的分子特征及其与胰岛素样雄性腺激素基因转录关系的研究

Molecular characterization of insulin-like androgenic gland hormone-binding protein gene from the oriental river prawn Macrobrachium nipponense and investigation of its transcriptional relationship with the insulin-like androgenic gland hormone gene.

作者信息

Li Fajun, Bai Hongkun, Xiong Yiwei, Fu Hongtuo, Jiang Sufei, Jiang Fengwei, Jin Shubo, Sun Shengming, Qiao Hui, Zhang Wenyi

机构信息

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, PR China; Weifang University of Science and Technology, Shouguang 262700, PR China.

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, PR China.

出版信息

Gen Comp Endocrinol. 2015 May 15;216:152-60. doi: 10.1016/j.ygcen.2014.12.007. Epub 2014 Dec 31.

Abstract

Insulin-like androgenic gland hormone-binding protein (IAGBP) has been investigated in crustaceans in vitro. However, the relationship between IAGBP and its putative binding protein partner insulin-like androgenic gland hormone (IAG) has not been studied at the transcriptional level in vivo. In the current study, we cloned the full-length cDNA of IAGBP from the oriental river prawn Macrobrachium nipponense (Mn-IAGBP) and investigated the transcriptional patterns of Mn-IAGBP and the M. nipponense IAG gene (Mn-IAG) at different developmental stages and in different tissues. Mn-IAGBP mRNA was detected in all examined tissues from adult male prawns, with the highest transcriptional levels in the testis. Mn-IAG mRNA was detected in the androgenic gland and hepatopancreas. The genomic sequences of Mn-IAGBP and Mn-IAG were isolated by genome walking and two gene copies were found in both Mn-IAGBP and Mn-IAG. The relationship between Mn-IAGBP and Mn-IAG at the transcriptional level was studied by RNA interference. Injection of Mn-IAGBP double-stranded RNA (dsRNA) significantly reduced the transcription of Mn-IAG, while injection of Mn-IAG dsRNA significantly reduced the transcription of Mn-IAGBP in testis, muscle, androgenic gland, and hepatopancreas. These results demonstrate the involvement of the IAGBP gene in IAG signaling in M. nipponense.

摘要

胰岛素样雄性腺激素结合蛋白(IAGBP)已在甲壳类动物中进行了体外研究。然而,IAGBP与其假定的结合蛋白伴侣胰岛素样雄性腺激素(IAG)之间的关系尚未在体内转录水平上进行研究。在本研究中,我们从日本沼虾(Macrobrachium nipponense)中克隆了IAGBP的全长cDNA(Mn-IAGBP),并研究了Mn-IAGBP和日本沼虾IAG基因(Mn-IAG)在不同发育阶段和不同组织中的转录模式。在成年雄性沼虾的所有检测组织中均检测到Mn-IAGBP mRNA,其中在睾丸中的转录水平最高。在雄性腺和肝胰腺中检测到Mn-IAG mRNA。通过基因组步移分离了Mn-IAGBP和Mn-IAG的基因组序列,并且在Mn-IAGBP和Mn-IAG中均发现了两个基因拷贝。通过RNA干扰研究了Mn-IAGBP和Mn-IAG在转录水平上的关系。注射Mn-IAGBP双链RNA(dsRNA)显著降低了Mn-IAG的转录,而注射Mn-IAG dsRNA显著降低了睾丸、肌肉、雄性腺和肝胰腺中Mn-IAGBP的转录。这些结果表明IAGBP基因参与了日本沼虾的IAG信号传导。

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