Department of Biology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran.
Department of Anatomy and Embryology, Infertility and Health Reproductive Research Center, Babol University of Medical Sciences, P.O. Box: 4136747176, Babol, Iran.
Reprod Sci. 2021 Apr;28(4):963-972. doi: 10.1007/s43032-020-00326-z. Epub 2021 Jan 25.
Several lines of evidence strongly suggest that retinoic acid (RA) and stem cell factor (SCF)/c-Kit signal transduction pathways are involved in the differentiation of spermatogonial stem cells (SSCs). This study was aimed to investigate the effect of RA and SCF on in vitro differentiation of SSCs via evaluation of the mRNA expression of meiosis-specific genes in cultured testicular tissues. Testicular tissue samples were obtained from bilaterally vasectomized rats and also healthy adult rats and then were cultured for 25, 30, and 35 days on different conditions. The cultured testicular pieces were sectioned and stained with PAS to histological analysis. The total RNA was extracted from cultured testicular samples, and the expression of ACR, PRTM1, SYCP3, STRA8, c-KIT, PIWIL2, and OCT4 genes at mRNA level was quantified using real-time polymerase chain reaction (qPCR) procedure. After 1-month surgery, bilateral testicular weight showed a significant decrease in vasectomized adult rats compared with healthy adult rats (P < 0.05). Reduction in the diameter of the seminiferous tubules and depletion of advanced germinal elements in vasectomized rats compared with healthy adult rats were also observed. Our findings also demonstrated that the mRNA expression level of PRTM1, STRA8, c-KIT, PIWIL2, and OCT4 genes in cultured testicular tissues significantly up-regulated in experimental group II compared with the control group (P < 0.001). Our findings lead us to conclude that SCF improves in vitro differentiation of SSCs in the OA rats, at least partially, by transcriptionally upregulating PRTM1, STRA8, c-KIT, PIWIL2, and OCT4 genes.
有几条证据线强烈表明，视黄酸（RA）和干细胞因子（SCF）/c-Kit 信号转导途径参与了精原干细胞（SSC）的分化。本研究旨在通过评估培养睾丸组织中减数分裂特异性基因的 mRNA 表达来研究 RA 和 SCF 对 SSC 体外分化的影响。从双侧输精管结扎大鼠和健康成年大鼠获得睾丸组织样本，然后在不同条件下培养 25、30 和 35 天。对培养的睾丸组织进行切片并用 PAS 染色进行组织学分析。从培养的睾丸组织中提取总 RNA，使用实时聚合酶链反应（qPCR）程序定量分析 ACR、PRTM1、SYCP3、STRA8、c-KIT、PIWIL2 和 OCT4 基因在 mRNA 水平的表达。手术后 1 个月，与健康成年大鼠相比，输精管结扎成年大鼠双侧睾丸重量明显下降（P<0.05）。与健康成年大鼠相比，结扎大鼠的生精小管直径减小，高级生殖细胞耗竭。我们的研究结果还表明，与对照组相比，实验组 II 中培养睾丸组织中 PRTM1、STRA8、c-KIT、PIWIL2 和 OCT4 基因的 mRNA 表达水平显著上调（P<0.001）。我们的研究结果表明，SCF 通过转录上调 PRTM1、STRA8、c-KIT、PIWIL2 和 OCT4 基因，至少部分改善了 OA 大鼠中 SSC 的体外分化。
