Expression of RSPH6A in the first wave of rat spermatogenesis and oxidative stress conditions: Attenuation by melatonin

Abstract

PurposeHere, we report, for the first time, the temporal expression and localization of axonemal radial spoke head homolog A (RSPH6A) protein during the first wave of rat spermatogenesis and in oxidative stress conditions.MethodsFor the developmental study, testes were collected from rats at different developmental stages (7, 14, 21, 28, 35, 42, and 60 postnatal days); for in vivo treatment, 24 rats were treated with cadmium and/or melatonin. From each sample, western blot (WB) and immunofluorescence (IF) analyses for RSPH6A were performed.ResultsRSPH6A expression starts at 21 PND alongside the appearance of I spermatocytes (SPC) with a significant increase up to 60 PND. Data were confirmed by IF analysis, showing that RPSH6A expression is restricted to I and II SPC, spermatids, and mature sperm. In vivo experiments showed that the expression and localization of RSPH6A in the testis and epididymal spermatozoa of adult rats treated with cadmium were impaired. Interestingly, melatonin (an antioxidant), given together with Cd, can counteract its damaging effects.ConclusionsAll combined data confirm that RSPH6A contributes to the onset of fertility by acting on sperm motility, raising the possibility of using RSPH6A as a marker for normal fertility in the general population.

Expression of Insl3 Protein in Adult Danio rerio

“…The samples were sonicated three times (20 Hz for 20 s each), placed on ice for 30 min, and then centrifuged at 10,000× g for 30 min at 4 • C. The supernatants were collected. Forty micrograms of the protein extracts was separated into SDS-PAGE (9-15% acrylamide) and treated according to Romano et al [41]. Then, they were incubated overnight at 4 • C with primary antibodies as follows: anti-INSL3 (1:800) and anti-α-Tubulin (1:5000, E-AB-20036, Elabscience ® , Houston, TX, USA).…”

Type 1 diabetes impairs the activity of rat testicular somatic and germ cells through NRF2/NLRP3 pathway-mediated oxidative stress

“…The major impact of oxidative stress on fertility has also been highlighted by the fact that many intervention strategies to face infertility are just addressed to reduce oxidative stress-for example, using antioxidant molecules (23)(24)(25)(26)(27)(28)(29)(30)(31). In this regard, our recent research focuses on the cellular and differentiative mechanisms that regulate spermatogenesis, which was perturbed using experimental models of oxidative stress (32,33), to expand the knowledge of the molecular pathways involved in spermatogenesis.…”