Zinc mitigates cadmium-induced sperm dysfunction through regulating Ca2+ and metallothionein expression in the freshwater crab Sinopotamon henanense.

Cadmium (Cd) is a highly toxic heavy metal element that might adversely affect sperm function such as the acrosome reaction (AR). Although it is widely recognized that zinc (Zn) plays a crucial role in sperm quality, the complete elucidation of how Zn ameliorates Cd-induced sperm dysfunction is still unclear. In this study, we aimed to explore the protective effects of Zn against the sperm dysfunction induced by Cd in the freshwater crab Sinopotamon henanense. The results demonstrated that Cd exposure not only impaired the sperm ultrastructure, but also caused sperm dysfunction by decreasing the AR induction rate, acrosome enzyme activity, and Ca2+ content in sperm while elevating the activity and transcription expression of key Ca2+ signaling pathway-related proteins Calmodulin (CAM) and Ca2+-ATPase. However, the administration of Zn was found to alleviate Cd-induced sperm morphological and functional disorders by increasing the activity and transcription levels of CaM and Ca2+-ATPase, thereby regulating intracellular Ca2+ homeostasis and reversing the decrease in Ca2+ contents caused by Cd. Furthermore, this study was the first to investigate the distribution of metallothionein (MT) in the AR of S. henanense, and it was found that Zn can reduce the elevated levels of MT in crabs caused by Cd, demonstrating the significance of Zn in inducing MT to participate in the AR process and in metal detoxification in S. henanense. These findings offer novel perspectives and substantiation regarding the utilization of Zn as a protective agent against Cd-induced toxicity and hold significant practical implications for mitigating Cd-induced sperm dysfunction.

Isolation, cloning, and tissue distribution and functional analysis of ShP-glycoprotein in the freshwater crab Sinopotamon henanense exposed to Cd and Cd-QDs.

P-glycoprotein (Pgp), a member of ATP binding cassette (ABC) transporter family, can extrude toxic substances out of cells by mediating multi-xenobiotic resistance (MXR) in aquatic organisms, however, its regulation and association with MXR are still unclear. In this work, the genetic information of Pgp in freshwater crab Sinopotamon henanense (ShPgp) was revealed for the first time. ShPgp with a total of 4488 bp was cloned and analyzed, which includes 4044 bp open reading frame, 353 bp 3' untranslated region, and 91 bp 5' untranslated region. The recombinant ShPGP were expressed in Saccharomyces cerevisiae and taken for SDS-PAGE and western blot analysis. ShPGP was widely expressed in the midgut, hepatopancreas, testis, ovary, gill, hemocytes, accessory gonad and myocardium of the crabs studied. The images of immunohistochemistry indicated that ShPgp was mainly distributed in the cytoplasm and cell membrane. When the crabs were exposed to cadmium or cadmium containing quantum dots (Cd-QDs), not only the relative expression of ShPgp mRNA and the protein produced were enhanced, but also the MXR activity and ATP contents. The relative expression of target genes related to energy metabolism, detoxification and apoptosis was also determined in the carbs exposed to Cd or Cd-QDs. The results showed that bcl-2 was significantly down-regulated, while other genes were up-regulated except PPAR (not affected). However, when the Shpgp in treated crabs was interfering by knockdown technique, their apoptosis and the expression of proteolytic enzyme genes and transcription factors MTF1 and HSF1 were also elevated, while the expression of apoptosis inhibiting and fat metabolism genes were compromised. Based on the observation, we concluded that MTF1 and HSF1 were involved in gene transcription regulation of mt and MXR, respectively, while PPAR had limited regulatory effect on those genes in S. henanense. NF-κB may play a negligible role in the process of apoptosis in testes induced by cadmium or Cd-QDs. However, the detail information regarding Pgp involvement in SOD or MT, and its association with apoptosis during xenobiotics insults remain to be explored.

Zinc alleviates cadmium-induced reproductive toxicity via regulating ion homeostasis, metallothionein expression, and inhibiting mitochondria-mediated apoptosis in the freshwater crab Sinopotamon henanense.

Cadmium (Cd) is a carcinogenic environmental pollutant that harms male reproductive systems by lowering sperm quality, impairing spermatogenesis, and causing apoptosis. Although zinc (Zn) has been reported to alleviate Cd toxicity, the underlying mechanisms have not been fully elucidated. The aim of this work was to investigate the mitigating effects of Zn on Cd-induced male reproductive toxicity in the freshwater crab Sinopotamon henanense. Cd exposure not only resulted in its accumulation but also in Zn deficiency, decreased sperm survival rate, poor sperm quality, altered ultrastructure, and increased apoptosis in the testis of the crabs. Morever, Cd exposure increased the expression and distribution of metallothionein (MT) in the testis. However, Zn supplementation effectively mitigated the aforementioned effects of Cd, as demonstrated by preventing Cd accumulation, increasing Zn bioavailability, alleviating apoptosis, increasing mitochondrial membrane potential, decreasing reactive oxygen species (ROS) levels, and restoring MT distribution. Moreover, Zn also significantly reduced the expression of apoptosis-related (p53, Bax, CytC, Apaf-1, Caspase-9, Caspase-3), metal transporter-related ZnT1, metal-responsive transcription factor 1 (MTF1), and the gene and protein expression of MT, while increasing the expression of ZIP1 and Bcl-2 in the testis of Cd-treated crabs. In conclusion, Zn alleviates Cd-induced reproductive toxicity via regulating ion homeostasis, MT expression, and inhibiting mitochondria-mediated apoptosis in the testis of S. henanense. The information obtained in this study may serve as the foundation for further investigation into the development of mitigation strategies for adverse ecological and human health outcomes associated with Cd contamination or poisoning.

Detoxification and recovery after cadmium exposure in the freshwater crab Sinopotamon henanense.

Cadmium (Cd) is a common pollutant in the aquatic environment, which puts the health and safety of aquatic organisms and humans at risk. In the present study, the freshwater crab Sinopotamon henanense was exposed to Cd (0, 50, 100, and 500 µg·L-1) for 14 d (0-14th d), followed by 21 d (14-35th d) of depuration. The changes in Cd bioaccumulation, microstructure, biomacromolecules (polysaccharides, neutral lipids, DNA and total proteins), and biochemical parameters (SOD, CAT, GR, TrxR, MDA and AChE) in the gills and hepatopancreas were tested. The injured microstructure, activated antioxidant system, increased MDA, and inhibited AChE of the gills and hepatopancreas responded with progressive bioaccumulation of Cd. Meanwhile, the polysaccharides and neutral lipids in the hepatopancreas reduced and DNA synthesis enhanced. During depuration, more than 58.80 ± 8.53% and 13.84 ± 12.11% of Cd was excreted from the gills and hepatopancreas, respectively. Recovery of microstructure and biomacromolecules as well as alleviated oxidative damage and neurotoxicity were also found in these two organs. Additionally, based on PCA, Ihis, GR and MDA were identified as the optimal biomarkers indicating the health status of crabs. In conclusion, S. henanense could resist Cd stress through antioxidant defence and self-detoxification.

Reproductive toxicity of quantum dots on gonads of the fresh water crab Sinopotamon henanense.

Since nano-quantum dots (QDs) are increasingly used as fluorescent dyes in biomedical sciences, the possibility of QDs contaminating aquatic environments is generally increasing. There is concern about potential toxicity of QDs. However, their risks in the aquatic environment are not entirely understood. In this study, the freshwater crab Sinopotamon henanense was exposed to cadmium telluride (CdTe) QDs by intraperitoneal injection to detect the reproductive toxicity of QDs (1/32, 1/16 and 1/4 LD50; Crab was exposed for 1, 3, 5, and 7 days). After CdTe QD exposure, no significant effect was detected on the body weight and gonadosomatic index. Additionally, morphological observations showed tissue vacuolation in the testis, and inflammatory cell infiltration in the ovary. The submicroscopic structure showed that exposure to CdTe QDs damaged the organelles and cell structures of the gonads of S. henanense. Among the adverse effects, pathological changes in the nuclear membrane, mitochondria and lysosomes were particularly significant. Antioxidant enzymes responded differently to different doses of QDs. The 0.5-mg/kg dose induced superoxide dismutase activity in the testes. And in the 1-mg/kg and 4-mg/kg dose QD exposure test, the testis responded by activating glutathione peroxidase and inducing reduced glutathione and overconsuming glutathione peroxidase. Respectively, the ovaries responded by overconsuming superoxide dismutase and glutathione peroxidase and reduced glutathione. Thus, we conclude that the gonads of S. henanense were injured by CdTe QD, and male are better indicators of the toxicity of QDs than female crabs according to greater alterations in tissue structure and antioxidant enzyme in the analyses.

Mitigative effects of zinc on cadmium-induced reproductive toxicity in the male freshwater crab Sinopotamon henanense.

Cadmium (Cd) is a highly harmful environmental contaminant, which can cause reproductive toxicity. Zinc (Zn) is an essential trance element that may protect the organism from the harmful effects of Cd. However, the mechanism of Zn against Cd-induced reproductive toxicity remained to be elucidated. The aim of this study was to assess the effects of subchronic exposure to Cd on the relative testis weight (RTW), the histopathology, the activity of stress marker antioxidant enzymes, the level of lipid peroxidation of testis, as well as the mitigative effects of Zn on Cd-induced reproductive toxicity in male freshwater crab Sinopotamon henanense. For this purpose, male crabs were divided into 10 groups including a control group (without metals) and metal exposure groups with Cd alone in three concentrations and Cd combined with Zn in six concentrations for 14 days. The results showed that Cd evoked concentration-dependent reproductive toxicity of male Sinopotamon henanense as showed by decreased RTW, appearance of morphological lesions, increased SOD, CAT, GPx activity, and MDA levels. Nevertheless, Zn combined with Cd exposure significantly alleviated Cd-induced reproductive toxicity as proved by increased RTW, reappearance of normal histological morphology, increased SOD activity, recovered CAT and GPx activity, and decreased MDA levels in testis. Our study demonstrated that the application of Zn can mitigate Cd-induced reproductive toxicity by ameliorating the testicular oxidative stress and improving the antioxidant status.