Sub-lethal concentrations of CdCl2 disrupt cell migration and cytoskeletal proteins in cultured mouse TM4 Sertoli cells

Abstract

The aims of this study were to examine the effects of CdCl2 on the viability, migration and cytoskeleton of cultured mouse TM4 Sertoli cells. Time-and concentration-dependent changes were exhibited by the cells but 1 µM CdCl2 was sub-cytotoxic at all time-points. Exposure to 1 and 12 µM CdCl2 for 4 h resulted in disruption of the leading edge, as determined by chemical staining. Cell migration was inhibited by both 1 and 12 µM CdCl2 in a scratch assay monitored by live cell imaging, although exposure to the higher concentration was associated with cell death. Western blotting and immunofluorescence staining indicated that CdCl2 caused a concentration dependent reduction in actin and tubulin levels. Exposure to Cd 2+ also resulted in significant changes in the levels and/or phosphorylation status of the microtubule and microfilament destabilising proteins cofilin and stathmin, suggesting disruption of cytoskeletal dynamics. Given that 1-12 µM Cd 2+ is attainable in vivo, our findings are consistent with the possibility that Cd 2+ induced impairment of testicular development and reproductive health may involve a combination of reduced Sertoli cell migration and impaired Sertoli cell viability depending on the timing, level and duration of exposure.

Cadmium‐induced toxicity increases prolyl endopeptidase (PREP) expression in the rat testis

“…In addition, the immunofluorescence analysis further corroborated the above result, showing quite stronger signal in Cd group, compared to others, not only in all the cells composing the germinal compartment, but also in the somatic components, Sertoli, and LC. According to the literature, Cd affected the normal cytoskeletal network in both germinal and somatic cells in the testis, and in particular, actin microfilaments (Siu et al, 2009) and microtubules (Egbowon, Harris, Arnott, Mills, & Hargreaves, 2016). Recently, we reported that expression of the formin DAAM1 decreased by Cd treatment (Chemek et al, 2018), reflecting damages to the cytoskeleton of GC.…”

“…Actually, PREP has also been associated to these cytoskeleton elements, suggesting engagement of the endopeptidase in microtubule‐associate processes, independent of its peptidase activity (Schulz et al, 2005), such as the cytoskeletal remodeling which occurs during GC movement. Cd toxicity alters distribution and dynamics of the microtubules network, inhibiting Sertoli cell migration and normal activity (Egbowon et al, 2016). Even in this second case, the enhanced PREP immunofluorescent signal in these cells may be of crucial significance to contrast the disorganization of microtubule network, which could be one of the major causes of GC loss and exfoliation from the epithelium.…”

Cry1 deficiency leads to testicular dysfunction and altered expression of genes involved in cell communication, chromatin reorganization, spermatogenesis, and immune response in mouse testis

“…These processes involve many genes whose dysregulation can lead to testicular dysfunction (Li, Lee, & Cheng, ; Skakkebaek et al, ). External stimuli such as toxicity, temperature change, and nutrient status have also been shown to influence spermatogenesis by altering the expression of genes in the testis (Cuypers et al, ; Egbowon, Harris, Arnott, Mills, & Hargreaves, ; Mocarelli et al, ; Skakkebaek et al, ; Spiazzi et al, ; Valles, Aveldano, & Furland, ). Identifying these genes can provide insight into the molecular basis of normal testicular function.…”

Effects of heavy metals on reproduction owing to infertility

“…Thus Cd 2+ toxicity damage sertoli cells by disarrangement of cytoskeletal network and other structural elements. [ 77 ] In rat uterus, Cd 2+ exposure induces oxidative stress, and accumulation of a high amount of malondialdehyde (MDA) produced by per oxidation of polyunsaturated fatty acid causes gynecological organ damage that may have a harmful impact on reproductive performance. [ 78 ] In male rats, Cd 2+ exposure increase glucocorticoid secretion by stimulating α‐adrenergic receptor and high amount of glucocorticoids inhibits the steroidogenesis in Leydig cells.…”

“…In mammals like rodents, Cd 2+ also induces cytotoxicity, edema, efferent duct necrosis, and testis necrosis by affecting signal transductions, such as PKC, cyclic AMP (cAMP), and MAPK pathway. [ 35,77 ] Cd 2+ exposure on testes causes lethal effects like the disruption of blood vessels, disassembly of BTB, seminiferous tubule damage, germ cell loss, low androgen secretion, testicular edema, hemorrhage, necrosis, infertility, and sterility. [ 16,35 ] In rodents, Cd 2+ toxicity also causes overstimulation of testicular cells by the stimulating pituitary gland, thereby increasing the production of LH production.…”