Chen Haolin, Pu Ling, Tian Chengcheng, Qi Xingcai, Song Juanjuan, Liao Yan, Mo Bentian, Li Taotao
Institute of Animal Husbandry and Veterinary, Guizhou Academy of Agricultural Sciences, Guiyang 550005, China.
College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Vet Sci. 2024 Jun 9;11(6):266. doi: 10.3390/vetsci11060266.
Platelet-derived growth factor B (PDGFB), as an important cellular growth factor, is widely involved in the regulation of cellular events such as cell growth, proliferation, and differentiation. Although important, the expression characteristics and biological functions in the mammalian reproductive system remain poorly understood. In this study, the PDGFB gene of Tibetan sheep was cloned by RT-PCR, and its molecular characteristics were analyzed. Subsequently, the expression of the PDGFB gene in the testes and epididymides (caput, corpus, and cauda) of Tibetan sheep at different developmental stages (3 months, 1 year, and 3 years) was examined by qRT-PCR and immunofluorescence staining. A bioinformatic analysis of the cloned sequences revealed that the CDS region of the Tibetan sheep PDGFB gene is 726 bp in length and encodes 241 amino acids with high homology to other mammals, particularly goats and antelopes. With the increase in age, PDGFB expression showed an overall trend of first decreasing and then increasing in the testis and epididymis tissues of Tibetan sheep, and the PDGFB mRNA expression at 3 months of age was extremely significantly higher than that at 1 and 3 years of age ( < 0.05). The PDGFB protein is mainly distributed in testicular red blood cells and Leydig cells in Tibetan sheep at all stages of development, as well as red blood cells in the blood vessel, principal cells, and the pseudostratified columnar ciliated epithelial cells of each epididymal duct epithelium. In addition, PDGFB protein expression was also detected in the spermatocytes of the 3-month-old group, spermatids of the 1-year-old group, spermatozoa and interstitial cells of the 3-year-old group, and loose connective tissue in the epididymal duct space in each developmental period. The above results suggest that the PDGFB gene, as an evolutionarily conserved gene, may play multiple roles in the development and functional maintenance of testicular cells (such as red blood cells, Leydig cells, and germ cells) and epididymal cells (such as red blood cells, principal cells, and ciliated epithelial cells) during testicular and epididymal development, which lays a foundation for the further exploration of the mechanisms by which the PDGFB gene influences spermatogenesis in Tibetan sheep.
血小板衍生生长因子B(PDGFB)作为一种重要的细胞生长因子,广泛参与细胞生长、增殖和分化等细胞活动的调节。尽管其很重要,但在哺乳动物生殖系统中的表达特征和生物学功能仍知之甚少。在本研究中,通过RT-PCR克隆了藏绵羊的PDGFB基因,并分析了其分子特征。随后,采用qRT-PCR和免疫荧光染色检测了不同发育阶段(3个月、1岁和3岁)藏绵羊睾丸和附睾(头、体和尾)中PDGFB基因的表达。对克隆序列的生物信息学分析表明,藏绵羊PDGFB基因的CDS区长度为726 bp,编码241个氨基酸,与其他哺乳动物,特别是山羊和羚羊具有高度同源性。随着年龄的增长,藏绵羊睾丸和附睾组织中PDGFB的表达呈现先下降后上升的总体趋势,3月龄时的PDGFB mRNA表达极显著高于1岁和3岁时(<0.05)。PDGFB蛋白主要分布于各发育阶段藏绵羊睾丸的红细胞和睾丸间质细胞,以及血管中的红细胞、附睾各段导管上皮的主细胞和假复层柱状纤毛上皮细胞。此外,在3月龄组的精母细胞、1岁组的精子细胞、3岁组的精子和间质细胞以及各发育时期附睾管腔隙中的疏松结缔组织中也检测到了PDGFB蛋白表达。上述结果表明,PDGFB基因作为一个进化保守基因,可能在睾丸和附睾发育过程中对睾丸细胞(如红细胞、睾丸间质细胞和生殖细胞)和附睾细胞(如红细胞、主细胞和纤毛上皮细胞)的发育和功能维持发挥多种作用,为进一步探究PDGFB基因影响藏绵羊精子发生的机制奠定了基础。