A brief exposure to cadmium impairs Leydig cell regeneration in the adult rat testis

Abstract

Cadmium is an endocrine disruptor, impairing male reproduction. The objective of this study is to investigate whether cadmium affects rat Leydig cell regeneration and to dissect the underlying mechanism. Adult male Sprague-Dawley rats received a single intraperitoneal injection (i.p.) of 0, 0.5 or 1.0 mg/kg of cadmium chloride, followed by ethane dimethane sulfonate (EDS) treatment to eliminate adult Leydig cells 20 days later. Compared to control (0 dose), cadmium treatment reduced serum testosterone levels by days 21, 35, and 56 after EDS treatment. Serum luteinizing hormone (LH) levels were also affected by day 56, the only time point examined. There were fewer regenerated Leydig cells in the cadmium-treated testis on days 35 and 56 after EDS treatment. Further studies demonstrated that the mRNA or protein levels of Leydig (Lhcgr, Scarb1, Star, Cyp11a1, Hsd3b1, Cyp17a1, Hsd17b3, and Hsd11b1), non-Leydig (Fshr and Dhh), and gonadotroph (Lhb) cells were also significantly lower in cadmium-treated animals. Since LH and desert hedgehog (DHH) are critical factors for Leydig cell differentiation, our result demonstrated that the lower doses of cadmium exposure, even briefly, may permanently damage Leydig cell regeneration.

Fibroblast growth factor 16 stimulates proliferation but blocks differentiation of rat stem Leydig cells during regeneration

“…Western blotting was carried out as previously described . Briefly, the testis or seminiferous tubules were homogenized in the lysis buffer (Bocai, Shanghai, China).…”

“…Testis and seminiferous tubule samples were homogenized in Trizol agent (Invitrogen, Carlsbad, CA, USA) for total RNA purification as previously described. 20 After extraction, the concentration of total RNA was measured by a plate reader at OD260. The first strand of cDNA was biosynthesized and used as the template for qPCR am- The steady-state levels of mRNAs were determined by a standard curve method.…”

“…Western blotting was carried out as previously described. 20 ACTB serves as a control. The protein levels were quantified using ImageJ software and normalized to ACTB.…”

Oncostatin M inhibits differentiation of rat stem Leydig cells in vivo and in vitro

“…The objective of the assay is to quest the influence of serum LH and FSH levels after the treatment of OSM. Serum LH and FSH levels were detected with enzyme linked immunosorbent assay (ELISA) kit according to the manufacturer's instructions (Chemicon, Temecula, CA, USA) as described previously . Briefly, an aliquot of a sample and assay diluent were added to each well of the pre‐coated 96‐well plate.…”

Dehydroepiandrosterone and Its CYP7B1 Metabolite 7α-Hydroxydehydroepiandrosterone Regulates 11β-Hydroxysteroid Dehydrogenase 1 Directions in Rat Leydig Cells

“…Western blotting was performed as previously described (29). Briefly, rat PLC, ILC, and ALC homogenates (30 µg protein each sample) were electrophoresed on 10% polyacrylamide gel containing sodium dodecyl sulfate.…”