Li Kunyu, Liu Yufang, He Xiaoyun, Tao Lin, Jiang Yanting, Lan Rong, Hong Qionghua, Chu Mingxing
Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
College of Life Science and Food Engineering, Hebei University of Engineering, Handan, China.
Front Cell Dev Biol. 2022 May 12;10:873095. doi: 10.3389/fcell.2022.873095. eCollection 2022.
, a member of the insulin-like growth factor (IGF) superfamily, is also known as the growth-promoting factor (somatomedin C). is involved in vertebrate growth and development, immunity, cell metabolism, reproduction, and breeding. However, there are relatively few studies on the relationship between and goat reproduction. In this study, a new transcription factor bound to the g. 64943050T>C promoted granulosa cell (GC) proliferation. A mutation g.64943050T>C located in the promoter region of was identified. Association analysis revealed that the kidding number in the first and second litters and the average number of first three litters of the CC genotype (2.206 ± 0.044, 2.254 ± 0.056, and 2.251 ± 0.031) were significantly higher than those in the TC genotype (1.832 ± 0.049, 1.982 ± 0.06, and 1.921 ± 0.034) and TT genotype (1.860 ± 0.090, 1.968 ± 0.117, and 1.924 ± 0.062) ( < 0.05). The kidding number in the third litter of the CC genotype (2.355 ± 0.057) was significantly higher than that in the TT genotype (2.000 ± 0.107) ( < 0.05). Then, the function of this mutation was validated by the dual-luciferase reporter assay and EMSA. The results showed that the luciferase activity of IGF1-mutant-C was significantly higher than that of IGF1-Wild-T ( < 0.05). The EMSA also showed that the binding ability of IGF1-mutant-C was higher than that of IGF1-Wild-T ( < 0.05). Subsequently, the transcription factor was predicted to bind to the mutation of (g.64943050T>C). Overexpression of SP1 promotes the expression of in the primary granulosa cells (GCs). The results of the CCK-8 assay and the expression of GC proliferation factors (, , and ) demonstrated that promoted GC proliferation by regulating expression. Our results suggested that the g.64943050T>C was significantly associated with the kidding number of Yunshang black goats, and as a transcription factor of binding to the mutation T>C regulated the expression of . Furthermore, promoted goat GC proliferation by regulating the expression of , which provides a new insight for the goat fertility trait.
胰岛素样生长因子（IGF）超家族的成员之一，也被称为生长促进因子（生长调节素C）。它参与脊椎动物的生长发育、免疫、细胞代谢、繁殖和育种。然而，关于它与山羊繁殖之间关系的研究相对较少。在本研究中，一种与IGF1基因g.64943050T>C结合的新转录因子促进了颗粒细胞（GC）增殖。在IGF1基因的启动子区域鉴定出一个g.64943050T>C突变。关联分析显示，CC基因型的第一胎和第二胎产羔数以及前三胎平均产羔数（2.206±0.044、2.254±0.056和2.251±0.031）显著高于TC基因型（1.832±0.049、1.982±0.06和1.921±0.034）和TT基因型（1.860±0.090、1.968±0.117和1.924±0.062）（P<0.05）。CC基因型第三胎产羔数（2.355±0.057）显著高于TT基因型（2.000±0.107）（P<0.05）。然后，通过双荧光素酶报告基因检测和电泳迁移率变动分析（EMSA）验证了该突变的功能。结果表明，IGF1 - 突变 - C的荧光素酶活性显著高于IGF1 - 野生 - T（P<0.05）。EMSA也表明，IGF1 - 突变 - C的结合能力高于IGF1 - 野生 - T（P<0.05）。随后，预测转录因子SP1与IGF1的突变（g.64943050T>C）结合。SP1的过表达促进了原代颗粒细胞（GCs）中IGF1的表达。CCK - 8检测结果和GC增殖因子（PCNA、Cyclin D2和Cyclin E）的表达表明，SP1通过调节IGF1表达促进了GC增殖。我们的结果表明，IGF1基因g.64943050T>C与云上黑山羊的产羔数显著相关，并且SP1作为与突变T>C结合的IGF1的转录因子调节了IGF1的表达。此外，SP1通过调节IGF1的表达促进了山羊GC增殖，这为山羊繁殖性状提供了新的见解。
