KISS1R and ANKRD31 Cooperate to Enhance Leydig Cell Gene Expression via the Cytoskeletal-Nucleoskeletal Pathway

Abstract

Kisspeptins are involved in the regulation of hypothalamic-pituitary-gonadal axis, Leydig cell functions, and testosterone secretion, acting as endogenous ligands of the KISS1 receptor. ANKRD31 protein participates in male fertility, regulating meiotic progression, and epididymal sperm maturation. Here, we show that in Leydig cells, KISS1 receptor and ANKRD31 proteins physically interact; the formation of this protein complex is enhanced by Kisspeptin-10 that also modulates F-actin synthesis, favoring histone acetylation in chromatin and gene expression via the cytoskeletal–nucleoskeletal pathway. Kp/KISS1R system deregulation, expression impairment of cytoskeletal–nucleoskeletal mediators, Leydig gene targets, and the decreased testosterone secretion in Ankrd31−/− testis strongly supported our hypothesis. Furthermore, cytochalasin D treatment subverted the gene expression induction dependent on Kisspeptin-10 action. In conclusion, the current work highlights a novel role for the Kisspeptin-10 in the induction of the cytoskeletal–nucleoskeletal route, downstream a physical interaction between KISS1 receptor and ANKRD31, with gene expression activation as final effect, in Leydig cells.

The interplay between kisspeptin and endocannabinoid systems modulates male hypothalamic and gonadic control of reproduction in vivo

“…In the hypothalamus, Kiss1 is notably produced in a sex-specific manner by neuronal populations mainly located within the hypothalamic ARC (both females and males) and the anteroventral-peri-ventricular nucleus (AVPV) (females only) to modulate GnRH pulse and mediate sex steroid feedbacks (5,6). The localization of Kiss1 and Kiss1R carried out in the present study confirmed that the ARC is the main brain region responsible for kisspeptin signaling in males, and the specific increase in the number of Kiss1+/Kiss1R+ neurons following Kp10 administration is consistent with the previously reported selfmodulatory loop of the KS (31,40,75,76). However, Western blot analysis of the basal hypothalamus did not show the same increase of Kiss1R found in the IHC of ARC.…”

“…Kp10 and AEA differentially modulated Kiss1R, Kiss1, and Cyp19 proteins, GnRH mRNA, and puberty-related miRs in the hypothalamus Kisspeptin is the main upstream modulator of GnRH, from puberty onset onward (5); both kisspeptin and estradiol (5,6,31,40), the sex steroid produced by the irreversible aromatization of testosterone by Cyp19 aromatase, were found as positive modulators of the KS. Conversely, the HPG axis is downregulated in CB1-/-mice (16,17).…”

“…Similarly, the involvement of the KS in Ca 2+ mobilization and SPZ physiology has been investigated in humans and mice (36,37) and is related to human sperm motility and hyperactivation (36) and fertilizing ability (37). Very recently, the involvement of Kiss1R in the cytoskeletal-nucleoskeletal pathway occurring in mouse Leydig cells to modulate gene expression was reported (40); in parallel, the possible role of the KS in epididymal SPZ maturation and storage was demonstrated in dogs (38) and rats (39), with significant Kiss1 levels measured within the epididymis (39). Based on the overall considerations, it appears that both ECS and KS centrally and locally regulate male reproduction; however, their possible interplay is still unraveled.…”

“…Despite the well-known role of the KS in the central control of reproduction, at the periphery, the expression of the KS has been reported in cell lines, testis, SPZ, and epididymis collected from several species (31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44), but the real functions of the KS in the testis and SPZ are far to be fully elucidated (8,45,46). In general, Leydig cells and germinal epithelium are the main sources and targets of Kiss1 in several species (8), and Kiss1 secretion from testis has been suggested (47).…”