Roles for Kisspeptin in proliferation and differentiation of spermatogonial cells isolated from mice offspring when the cells are cocultured with somatic cells

Abstract

Kisspeptin (Kp) expression in testis has caused most of the recent research surveying its functional role in this organ. This peptide influences spermatogenesis and sperm capacitation, so it is considered as a regulator of reproduction. Kp roles exert through hypothalamic/pituitary/gonadal axis. We aimed to evaluate direct roles for Kp on proliferation and differentiation of spermatogonial cells (SCs) when the cells are cocultured with somatic cells. Somatic cells and SCs were isolated from adult azoospermic and newborn mice and then enriched using a differential attachment technique. After the evaluation of identity and colonization for SCs, the cells were cocultured with somatic cells, and three doses of Kp (10−8‐10−6 M) was assessed on proliferation (through evaluation of MVH and ID4 markers) and differentiation (via evaluation of c‐Kit and SCP3, TP1, TP2, and, Prm1 markers) of the coculture system. Investigations were continued for four succeeding weeks. At the end of each level of testosterone in the culture media was also evaluated. We found positive influence from Kp on proliferative and differentiative markers in SCs cocultured with somatic cells. These effects were dose‐dependent. There was no effect for Kp on testosterone level. From our findings, we simply conclude that Kp as a neuropeptide for influencing central part of reproductive axis could also positively affect peripheral processes related to spermatogenesis without having an effect on steroidogenesis.

Kisspeptin and Testicular Function—Is It Necessary?

“…Localisation of KISS1/KISS1R system in germ cells postulates its role in germ cell differentiation. Kp10, in a dose-dependent manner, positively stimulated, proliferative (ID4 and MVH) and differentiative markers (SCP3 and c-Kit) in spermatogonial cells from adult azoospermic and new-born mice cocultured with somatic cells [48]. Additionally, the effect of kisspeptin administration on accessory gland structure, seminal vesicle and prostate have also been documented [69].…”

“…These include acceleration of germ cell progression in fish [26], increased spermiation [43]. and alteration of oestradiol signaling in amphibians [24], advancement of germ cell proliferation [48]. in mice, to modulation of intracellular Ca 2+ in mice and humans [16,20].…”

“…• kp10: intracellular Ca 2+ in spermatozoa [20], proliferation and differentiation markers on spermatogonial cells [48]. • kp: role in sperm capacitation [20].…”

“…Arrows. 1: Increased intracellular calcium in spermatozoa, increased sperm motility and hyperactivation in men[16]; 2: Increased spermatogonial cell proliferation and differentiation markers in adult mice[48]; 3: Increased number of seminiferous tubules with detached spermatozoa in frogs[43]; 4: Nil effect of kp on testosterone secretion with or without cotreatment with LH or hCG in mice[12,42]; 5. Continuous infusions of kp led to seminiferous tubule degeneration and apoptosis, and reduction in, L.H.…”

Pentoxifylline improves the survival of spermatogenic cells via oxidative stress suppression and upregulation of PI3K/AKT pathway in mouse model of testicular torsion-detorsion

“…GAPDH served as a positive control for protein loading, and a high range molecular weight standard was used in determining the protein sizes. Immunoreactivity and signals were monitored by enhanced chemiluminescence [ 36 ] and the results were evaluated with the aid of NIH Image J software.…”