精子发生相关CG14片段的组织特异性表达及其与杂交不育的关系

目的 :确定精子发生相关 CG1 4片段的组织特异性表达及其与杂交不育的初步关系。方法 :采用 CG1 4片段的放射性同位素标记探针对心、肝、肾、脑、睾丸及附睾组织的 m RNA进行 Northern杂交。将 CG1 4片段转录为 RNA作探针 ,对成年雄性 SD大鼠、牦牛、犏牛一代 (杂交不育 )、犏牛三代 (回交生育功能恢复 )睾丸组织进行原位杂交。结果 :在睾丸组织有一条约 1 2 58bp的 m RNA特异杂交带 ;在附睾组织有一条约 1 3 51 bp的 m RNA特异杂交带 ,而在心、肝、肾及脑无杂交带。在 SD大鼠、牦牛及犏牛三代的睾丸组织冰冻切片存在明显的 CG1 4正义探针阳性杂交信号 ;在犏牛一代睾丸组织未见特异性阳性杂交信号 ;在各个组织切片中反义探针均未出现阳性杂交信号。结论 :精子发生相关 CG1 4片段在睾丸及附睾组织中特异性表达。在 SD大鼠、具有生育能力的牦牛以及生育机能恢复的犏牛三代初级精母细胞中表达 CG1 4片段的 m RNA,而在杂种不育的犏牛一代初级精母细胞中未见 CG1 4片段的 m RNA。

Tissue-specific expression of a spermatogenesis related CG14 fragment and its involvement in hybrid infertility

Objective: To determine the tissue specific expression of a spermatogenesis related CG14 fragment and its involvement in hybrid infertility Methods: Northern hybridization with CG14 probe was used to testify the expression of CG14 fragment in cardiac, liver, kidney, brain, testis, and epididymis tissues of rat. In situ hybridization technique was used to detect CG14 positive signals in testes of rat, yak as well as its crossbred filial generations. Results: The results of Northern blot hybridization with CG14 probe showed that one mRNA specific hybridization band with a size of 1 258 bp was found in testis tissue and another one with a size of 1 351 bp was present in epididymis tissue. However, there was no any expression of CG14 fragment in other tissues. The results of in situ hybridization exhibited that positive signals were present in the primary spermatocyte of rat and yak, but no positive signal existed in that of yak crossbred filial generaton which is infertile. Conclusions: 1. The CG14 gene fragment was expressed significantly in testes and epididymides of rat, but never expressed in cardiac, liver, kidney, and brain tissues. 2. The CG14 gene fragment was specifically present in the primary spermatocyte of rat as well as yak and its fertile offspring, but never existed in that of infertile crossbred filial generation.