Juneja Subhash C
Department of Zoology, The University of Western Ontario, London, Canada.
Indian J Physiol Pharmacol. 2003 Apr;47(2):147-56.
Gap junctions are made of connexin (Cx) molecules and provide communications between adjacent cells through which small molecules (<1kDa) move between cytoplasms. The connexin gene family consists of at least 17 members. Gap junctions play important roles in ovarian and testicular function. Connexin 43 (Cx43) knockout mouse is a model for developmentally impaired fetal gonads. The aim of the current study was twofold: (a) to analyze mRNA expression pattern of multiple members of connexin gene family in normal fetal gonads; (b) to investigate any alteration of mRNA expression of connexins in developmentally impaired fetal gonads. The study was conducted in normal gonads obtained from 17.5 dpc wildtype (Cx43+/+) and heterozygote (Cx43+/-) fetuses and in developmentally impaired gonads obtained from 17.5 dpc knockout (Cx43-/-) fetuses. The mRNA expression pattern of connexins (Cx26, Cx30.3, Cx31, Cx31.1, Cx32, Cx37, Cx40, Cx43, Cx45, Cx46 and Cx50) was analyzed by RT-PCR. The mRNA transcripts for Cx32 and Cx50 were absent in fetal testis of all the genotypes, and the transcripts for Cx26, Cx30.3, Cx31.1, Cx32, Cx40, Cx46 and Cx50 were absent in fetal ovary of all the genotypes. The transcripts for Cx43 showed expression in Cx43+/+ and Cx43+/- gonads and were absent in Cx43-/- gonads, as expected. Additionally, the mRNA transcripts for 8 more connexins (Cx26, Cx30.3, Cx31, Cx31.1, Cx37, Cx40, Cx45 and Cx46) showed expression in Cx43+/+ and Cx43+/- fetal testes, but the transcripts for only 4 connexins (Cx26, Cx37, Cx40 and Cx45) showed expression in Cx43-/- fetal testis. In fetal ovary, the mRNA transcripts for 3 more connexins (Cx31, Cx37 and Cx45) were expressed in all the genotypes. In summary, from the mRNA expression analysis of 11 members of connexin, gene family in 17.5 dpc fetal gonads, besides the expression of Cx43, 8 additional connexins were expressed in normal fetal testis but only 4 connexins were expressed in developmentally impaired testis. The fetal ovary showed the expression of 3 additional connexins besides the expression of Cx43. In developmentally impaired fetal ovary, only Cx43 was not expressed as expected but other three connexins were expresed. The study may be useful in interpreting human testis defects in infertility cases.
间隙连接由连接蛋白(Cx)分子构成,可在相邻细胞间提供通讯,小分子(<1kDa)可通过其在细胞质之间移动。连接蛋白基因家族至少由17个成员组成。间隙连接在卵巢和睾丸功能中发挥重要作用。连接蛋白43(Cx43)基因敲除小鼠是胎儿性腺发育受损的模型。本研究的目的有两个:(a)分析连接蛋白基因家族多个成员在正常胎儿性腺中的mRNA表达模式;(b)研究发育受损胎儿性腺中连接蛋白mRNA表达的任何变化。该研究在取自17.5天胚龄野生型(Cx43+/+)和杂合子(Cx43+/-)胎儿的正常性腺以及取自17.5天胚龄基因敲除(Cx43-/-)胎儿的发育受损性腺中进行。通过逆转录聚合酶链反应(RT-PCR)分析连接蛋白(Cx26、Cx30.3、Cx31、Cx31.1、Cx32、Cx37、Cx40、Cx43、Cx45、Cx46和Cx50)的mRNA表达模式。所有基因型胎儿睾丸中均未检测到Cx32和Cx50的mRNA转录本,所有基因型胎儿卵巢中均未检测到Cx26、Cx30.3、Cx31.1、Cx3十二条、Cx40、Cx46和Cx50的转录本。正如预期的那样,Cx43的转录本在Cx43+/+和Cx43+/-性腺中表达,而在Cx43-/-性腺中未表达。此外,另外8种连接蛋白(Cx26、Cx30.3、Cx31、Cx31.1、Cx37、Cx40、Cx45和Cx46)的mRNA转录本在Cx43+/+和Cx43+/-胎儿睾丸中表达,但只有4种连接蛋白(Cx26、Cx37、Cx40和Cx45)的转录本在Cx43-/-胎儿睾丸中表达。在胎儿卵巢中,另外3种连接蛋白(Cx31、Cx37和Cx45)的mRNA转录本在所有基因型中均有表达。总之,通过对17.5天胚龄胎儿性腺中11种连接蛋白基因家族成员的mRNA表达分析,除Cx43的表达外,正常胎儿睾丸中还表达另外8种连接蛋白,而在发育受损睾丸中仅表达4种连接蛋白。胎儿卵巢除Cx43的表达外,还表达另外3种连接蛋白。在发育受损的胎儿卵巢中,只有Cx43未如预期表达,但其他三种连接蛋白有表达。该研究可能有助于解释不育病例中人类睾丸缺陷。