Melatonin promotes development of haploid germ cells from early developing spermatogenic cells of Suffolk sheep under in vitro condition

Abstract

Promotion of spermatogonial stem cell (SSC) differentiation into functional sperms under in vitro conditions is a great challenge for reproductive physiologists. In this study, we observed that melatonin (10−7 m) supplementation significantly enhanced the cultured SSCs differentiation into haploid germ cells. This was confirmed by the expression of sperm special protein, acrosin. The rate of SSCs differentiation into sperm with melatonin supplementation was 11.85 ± 0.93% which was twofold higher than that in the control. The level of testosterone, the transcriptions of luteinizing hormone receptor (LHR), and the steroidogenic acute regulatory protein (StAR) were upregulated with melatonin treatment. At the early stage of SSCs culture, melatonin suppressed the level of cAMP, while at the later stage, it promoted cAMP production. The similar pattern was observed in testosterone content. Expressions for marker genes of meiosis anaphase, Dnmt3a, and Bcl‐2 were upregulated by melatonin. In contrast, Bax expression was downregulated. Importantly, the in vitro‐generated sperms were functional and they were capable to fertilize oocytes. These fertilized oocytes have successfully developed to the blastula stage.

Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

“…However, we found that melatonin can promote the primary SSC colony formation and promote the proliferation markers expression, this indicated that melatonin may affect goat SSCs indirectly. Interestingly, we also found that melatonin can inhibit the expression of SSC differentiation markers expression, which was contrary to the recently report that melatonin can promote the development of sperm from early developing spermatogenic cells of Suffolk sheep [39]. Here, we speculated melatonin may function though different ways to regulate SSCs when cultured in different medium.…”

Insights into in vitro spermatogenesis in mammals: Past, present, future

“…Successful in vitro differentiation of sheep SSCs into haploid sperm‐like cells was achieved using melatonin (Deng et al, ). This outcome is consistent with the crucial function of melatonin in differentiation along the spermatogenic lineage (Deng et al, ; Sanchez‐Barcelo et al, ; Tsantarliotou et al, ).…”

“…Kanatsu‐Shinohara and co‐workers cultured spermatogonia cells from neonatal mouse testes on a feeder layer of mouse embryonic fibroblasts, in the presence of various factors—such as GDNF, FGF2, Epidermal growth factor (EGF), Leukemia inhibitory factor (LIF), and fetal calf serum—but could only produce mature sperm cells when they transplanted the obtained cells into the testes of infertile mice (Kanatsu‐Shinohara et al, ). In another study using testicular cells from sheep, melatonin was used to induce differentiation of SSCs (Deng et al, ). Melatonin is synthesized in the testes and plays a role in testis function by increasing testosterone levels and modifying the morphology of spermatogenic cells (Sanchez‐Barcelo et al, ; Tsantarliotou, Kokolis, & Smokovitis, ).…”

“…Melatonin is synthesized in the testes and plays a role in testis function by increasing testosterone levels and modifying the morphology of spermatogenic cells (Sanchez‐Barcelo et al, ; Tsantarliotou, Kokolis, & Smokovitis, ). Deng and co‐workers reported that melatonin increased the rate of SSC differentiation twofold and up‐regulated the expression of differentiation‐related genes (Deng et al, ). Furthermore, the functional haploid sperm‐like cells produced could activate eggs, and yielded a high frequency of blastocysts—but no information about the ability of embryos to produce healthy offspring was provided.…”

Protective effects of melatonin and ghrelin on spermatogenesis: A narrative review of the literature

“…From then on, there have been a few types of research in this application of its. The most recent study, shows that administration of melatonin results in differentiation of SSCs to haploid germ cells in vitro, other than the advantages mentioned before (61). Gholami and coworkers had shown this advantage through the Sertoli cell modeling of seminiferous tubes induced by 40 mg/kg busulfan (62).…”