Combined effect of microplastic and triphenyltin: Insights from the gut-brain axis.

Microplastics (MPs), an emerging group of pollutants, not only have direct toxic effects on aquatic organisms but also cause combined toxicity by absorbing other pollutants. Triphenyltin (TPT), one of the most widely used organotin compounds, has adverse effects on aquatic organisms. However, little is known about the combined toxicity of MPs and TPT to aquatic organisms. To investigate the individual and combined toxicity of MPs and TPT, we selected the common carp (Cyprinus carpio) for a 42-day exposure experiment. Based on the environmental concentrations in a heavily polluted area, the experimental concentrations of MPs and TPT were set at 0.5 mg L-1 and 1 µg L-1, respectively. The effects of MPs combined with TPT on the carp gut-brain axis were evaluated by detecting gut physiology and biochemical parameters, gut microbial 16S rRNA, and brain transcriptome sequencing. Our results suggest that a single TPT caused lipid metabolism disorder and a single MP induced immunosuppression in carp. When MPs were combined with TPT, the involvement of TPT amplified the immunotoxic effect induced by MPs. In this study, we also explored the gut-brain axis relationship of carp immunosuppression, providing new insights for assessing the combined toxicity of MPs and TPT. At the same time, our study provides a theoretical basis for evaluating the coexistence risk of MPs and TPT in the aquatic environment.

Assessing the ecotoxicity of combined exposure to triphenyltin and norfloxacin at environmental levels: A case study of immunotoxicity and metabolic regulation in carp (Cyprinus carpio).

This paper evaluates the coexistence risks of triphenyltin (TPT) and norfloxacin (NOR) to aquatic organisms in the aquatic environment. Carp (Cyprinus carpio) was used as the test organism, the control and exposure groups (1 µg/L TPT), 1 mg/L (NOR), 1 µg/LTPT+1 mg/LNOR (TPT_NOR)) were set up according to the environmental concentration in the severely polluted area for 42 days. The single/combined toxic effects of TPT and NOR on aquatic organisms were evaluated by analyzing carp brain transcriptome sequencing, gut microbiota structure, and detection of biochemical indicators and RT-qPCR. Our results show that TPT and NOR induce lipid metabolism disorder in carp brain tissue, affecting the metabolism of cytochrome P450 to exogenous substances, and NOR also induces immunosuppression in carp. Long-term exposure to TPT combined with NOR amplifies the monotoxicity of TPT or NOR on lipid metabolism and immunosuppression in carp, induces immune dysfunction in brain tissue and changes in gut microbiota structure. However, TPT_NOR has no obvious neurotoxicity on the brain, but it can inhibit the level of intestinal MDA. This highlights that co-exposure of TPT and NOR amplifies metabolic disorders and immunosuppressive functions in carp.

Relationship between Water Temperature and Floating Time of Aquatic Cadavers.

OBJECTIVES: To study the relationship between water temperature and floating time of aquatic cadavers, providing a reference for more precise positioning and searching for floating corpses. METHODS: The floating model of guinea pig after drowning at 17-30 ℃ was established, and the floating times of carcasses were recorded. The collected data of 32 floating corpse cases in the Pearl River were sorted out and analyzed according to the floating time of corpses corresponding to each degree of water temperature. The relationship models between water temperature and the floating time of guinea pig carcass, and between that and the floating time of real cases were established. RESULTS: The floating time of the cadaver was negatively correlated with water temperature. The power function fitting equation of the relationship between floating time and water temperature of guinea pig carcass was y=1×1015x-10.530(R2=0.871, P<0.01), and the power function fitting equation of the relationship between corpse floating time and water temperature was y=3×106x-3.467(R2=0.802, P<0.01). CONCLUSIONS: It is found that average floating cadaver time has a power function with water temperature, which provides a reference for locating floating cadavers and establishing search models.

Exposure to enrofloxacin and depuration: Endocrine disrupting effect in juvenile grass carp (Ctenopharyngodon idella).

This study aimed to determine the effects of Enrofloxacin (ENR) exposure and depuration on the disruption of thyroid function and growth of juvenile grass carp (Ctenopharyngodon idella) as well as to assess the risk of ENR exposure to human health. Juvenile grass carp were treated with ENR solutions at different concentration gradients for 21 days and then depurated for 14 days. The results indicated ENR accumulation in the juvenile grass carp muscles, which persisted after depuration. In addition, exposure to ENR could alter growth by regulating the expression of genes associated with growth hormone/insulin-like growth factor (GH)/IGF) axis and the hypothalamic-pituitary-thyroid (HPT) axis. During ENR exposure, no significant changes in growth hormone levels were observed; however, a significant increase in the growth hormone level was noted. GH/IGF axis-related genes were upregulated after ENR exposure, and their expression levels remained high after depuration. Notably, a significant increase in the serum triiodothyronine (T3) and thyroxine (T4) levels coincided with the upregulation of HPT axis-related genes in both exposure and depuration treatments, and their expression levels remained high after depuration. Therefore, juvenile grass carp exposure to ENR induces physiological stress through HPT and GH/IGF axes that cannot be recovered after depuration. ENR accumulates in the muscles of juvenile grass carp and may pose a threat to human health. Therefore, exposure of juvenile grass carp to ENR results in impaired thyroid function and impaired growth. In addition, consumption of ENR-exposed fish poses human health risks.

Effects of long-term exposure of norfloxacin on the HPG and HPT axes in juvenile common carp.

Currently, there is a relatively lack of relevant research on the interference effect of quinolone antibiotics on the endocrine of aquatic animals. In this study, the toxicity of norfloxacin (NOR) on the endocrine system of juvenile common carp (Cyprinus carpio) was evaluated, as well as the hematocyte parameters. Specifically, two important endocrine axes were assessed: the hypothalamus-pituitary-thyroid (HPT) axis and hypothalamus-pituitary-gonadal (HPG) axis. Norfloxacin was used as a representative of quinolone antibiotics. According to the concentration of water pollution areas and considering the bad situation that may be caused by wastewater discharge, a control, 100 ng/L NOR, and 1 mg/L NOR treatment groups were set up. The juvenile carp, as the test animal, was subjected to an exposure experiment for 42 days. Thyroid hormones (T3 and T4) and related genes in HPT axis and sex hormones (11-ketotestosterone [11-KT] and progesterone [PROG]) and related genes in HPG axis and blood count are tested. It was found that the T4 iodine level and conversion process were enhanced after NOR treatment, which in turn led to the increase of T3 content and biological activity in the blood. One hundred nanograms per liter NOR can inhibit the level of sex hormones and inhibit the expression of HPG axis-related genes. In the 1 mg/L NOR treatment group, long-term exposure over a certain concentration range may lead to the development of adaptive mechanisms, making the changes in hormones and related genes insignificant. In conclusion, this study provides reference data for the endocrine interference of quinolone antibiotics on aquatic organisms, and has ecological significance for assessing the health of fish populations of quinolone antibiotics. However, the specific sites and mechanisms of action related to the effects of NOR on the endocrine system remain unclear and require further study.

Hepatotoxicity in carp (Cyprinus carpio) exposed to environmental levels of norfloxacin (NOR): Some latest evidences from transcriptomics analysis, biochemical parameters and histopathological changes.

Recently, the residues of quinolones have received widespread attention. However, toxicological studies on aquatic organisms are relatively scarce, especially on the liver metabolism and immune effects of these aquatic organisms. In this study, we investigated the toxic effects of carp exposed to 0, 100 ng/L, and 1 mg/L norfloxacin (NOR) at environmental concentrations for 42 days. In this study, through transcriptomics analysis, we found that some genes involved in lipid metabolism, immune response, and cytochrome P450 metabolism, especially genes accounting for the metabolism-related disorders of glucose and lipid. Defects in these genes and thus their related pathways increase the risk of coming down with nonalcoholic fatty liver disease. Compared to those of the control, results from the biochemical indicators of the treatment group changed significantly, including levels of total cholesterol, triglycerides, glucose, and insulin. Moreover, our results confirmed that NOR at environmental concentrations disordered the metabolism of glucose and lipid in the carp also resulted in hepatocellular and nuclear enlargement. Our results, therefore, confirmed that long-term exposure to NOR can induce carp liver toxicity at histological, biochemical, and transcriptional levels and provided the latest data and theoretical basis for the toxicology study of quinolones in the natural environment.

Effects of environmental norfloxacin concentrations on the intestinal health and function of juvenile common carp and potential risk to humans.

Antibiotics are emerging pollutants in our environment. These treatments have been widely used for their low cost, convenient use, and prominent effects. However, the prolonged or excessive use of such drugs can cause toxicity in aquatic organisms. These effects include genotoxicity, metabolic alteration, delayed development and decreased immunity, which carry further risks for ecological systems. In the present study, juvenile common carp (Cyprinus carpio) were exposed to norfloxacin (NOR) for 42 days, with NOR concentrations ranging from 100 ng/L to 1 mg/L, to assess the effects of environmental concentrations of antibiotics, to investigate the effects of NOR on intestinal morphology, enzymatic activity, and transcriptomic levels of RNA in fish, as well as a risk assessment on human health was carried out. The results demonstrated that oxidative stress was induced, the barrier function of the intestine was damaged, and changes occurred in the expression of immune-related genes in fish chronically exposed to antibiotics. Moreover, NOR could affect the regulation of the NF-κB signaling pathway. Thus, environmental concentrations of antibiotics can influence the intestinal health of fish and potentially posing health risks to humans.

A teleost bactericidal permeability-increasing protein-derived peptide that possesses a broad antibacterial spectrum and inhibits bacterial infection as well as human colon cancer cells growth.

The bactericidal permeability-increasing protein (BPI) is a multifunctional cationic protein produced by neutrophils with antibacterial, antitumor, and LPS-neutralizing properties. In teleost, a number of BPIs have been reported, but their functions are very limited. In this study, an N-terminal peptide, BO18 (with 18 amino acids), derived from rock bream (Oplegnathus fasciatus) BPI, was synthesized and investigated for its antibacterial spectrum, action mechanism, immunoregulatory property as well as the inhibition effects on bacterial invasion and human colon cancer cells growth. The results showed that BO18 was active against Gram-positive bacteria Bscillus subiilis, Micrococcus luteus, and Staphylococcus aureus, as well as Gram-negative bacteria Vibrio alginolyticus, Vibrio litoralis, Vibrio parahaemolyticus and Vibrio vulnificus. BO18 treatment facilitated the bactericidal process of erythromycin and rifampicin by enhancing the permeability of the outer membrane. During its interaction with V. alginolyticus, BO18 exerted its antibacterial activity by destroying cell membrane integrity, penetrating into the cytoplasm and binding to genomic DNA and total RNA. In vitro analysis indicated BO18 could enhance the respiratory burst ability and regulate the expression of immune related genes of macrophages. In vivo detection showed the administration of fish with BO18 before bacterial infection significantly reduced pathogen dissemination and replication in tissues. In addition, BO18 exerted a cytotoxic effect on the growth of human colon cancer cells HT-29. Together, these results add new insights into the function of teleost BPIs, and support that BO18 is a novel and broad-spectrum antibacterial peptide with potential to apply in fighting pathogenic infection in aquaculture.

Pachymic acid protects oocyte by improving the ovarian microenvironment in polycystic ovary syndrome mice†.

Women with polycystic ovary syndrome (PCOS) are characterized by endocrine disorders accompanied by a decline in oocyte quality. In this study, we generated a PCOS mice model by hypodermic injection of dehydroepiandrosterone, and metformin was used as a positive control drug to study the effect of pachymic acid (PA) on endocrine and oocyte quality in PCOS mice. Compared with the model group, the mice treated with PA showed the following changes (slower weight gain, improved abnormal metabolism; increased development potential of GV oocytes, reduced number of abnormal MII oocytes, and damaged embryos; lower expression of ovarian-related genes in ovarian tissue and pro-inflammatory cytokines in adipose tissue). All these aspects show similar effects on metformin. Most notably, PA is superior to metformin in improving inflammation of adipose tissue and mitochondrial abnormalities. It is suggested that PA has the similar effect with metformin, which can improve the endocrine environment and oocyte quality of PCOS mice. These findings suggest that PA has the similar effect with metformin, which can improve the endocrine environment and oocyte quality of PCOS mice.

Identification and characterization of a cathepsin K homologue that interacts with pathogen bacteria in black rockfish, Sebastes schlegelii.

Cathepsin K belongs to the family of cysteine cathepsins. It is well known that the cysteine cathepsins participate in various physiological processes and host immune defense in mammals. However, in teleost fish, the function of cathepsin K is very limited. In the present study, a cathepsin K homologue (SsCTSK) from the teleost black rockfish (Sebastes schlegelii) was identified and examined at expression and functional levels. In silico analysis showed that three domains, including signal peptide, cathepsin propeptide inhibitor I29 domain, and functional domain Pept_C1, are existed in SsCTSK. SsCTSK also possesses a peptidase domain with three catalytically essential residues (Cys25, His162 and Asn183). Phylogenetic profiling indicated that SsCTSK was evolutionally close to the cathepsin K of other teleost fish. Expression of SsCTSK occurred in multiple tissues and was induced by bacterial infection. Purified recombinant SsCTSK (rSsCTSK) exhibited apparent maximal peptidase activity at 45 °C, and its enzymatic activity was remarkably declined in the presence of the cathepsin inhibitor E-64. Moreover, rSsCTSK possesses the ability to bind with PAMPs and bacteria. Finally, knockdown of SsCTSK expression facilitated bacterial invasion in black rockfish. Collectively, these results indicated that SsCTSK functions as a cysteine protease and may serves as a target for pathogen manipulation of host defense system.

Effects of fine particulate matter (PM2.5) on ovarian function and embryo quality in mice.

Fine particulate matter (PM2.5) has an adverse effect on reproductive function, in particular causing reduced male reproductive function, but relatively few studies have directly targeted its effects on female reproduction. To investigate the effects of PM2.5 exposure on female reproduction, we exposed female mice to PM2.5 by intratracheal instillation for 28 days, and evaluated apoptosis of ovarian granulosa cells and oocytes and the quality embryos after insemination. Our results showed increased numbers of apoptotic granulosa cells and oocytes after exposure to elevated concentrations of PM2.5, which had adverse effects on female fertility via compromising embryo development and quality. We conclude that PM2.5 induced apoptosis of ovarian granulosa cells and oocytes leading to disrupted embryo development and female fertility in mice.

Characterization, expression, enzymatic activity, and functional identification of cathepsin S from black rockfish Sebastes schlegelii.

Cathepsin S belong to the cathepsin L-like family of cysteine cathepsins. It is well known that Cathepsin S participate in various physiological processes and host immune defense in mammals. However, in teleost fish, the function of cathepsin S is less investigated. In the present study, a cathepsin S homologue (SsCTSS) from the teleost fish black rockfish (Sebastes schlegelii) were identified and examined at expression and functional levels. In silico analysis showed that three domains, including signal peptide, cathepsin propeptide inhibitor I29 domain, and functional domain Pept_C1, were existed in the cathepsin. SsCTSS possesses a peptidase domain with three catalytically essential residues (Cys25, His162, and Asn183). Phylogenetic profiling indicated that SsCTSS are evolutionally close to the cathepsin S of other teleost fish. The expression of SsCTSS in immune-related tissues was upregulated in a time-dependent manner upon bacterial pathogen infection. Purified recombinant SsCTSS (rSsCTSS) exhibited apparent peptidase activity, which was remarkably declined in the presence of the cathepsin inhibitor E-64. rSsCTSS showed strong binding ability to LPS and PGN, the major constituents of the outer membranes of Gram-negative and Gram-positive bacteria, respectively. rSsCTSS also exhibited the capability of agglutination to different bacteria. The knockdown of SsCTSS attenuated the ability of host to eliminate pathogenic bacteria. Taken together, our results suggested that SsCTSS functions as cysteine protease which might be involved in the antibacterial immunity of black rockfish.

The effects of graphene quantum dots on the maturation of mouse oocytes and development of offspring.

Recently, graphene nanomaterials have attracted tremendous attention and have been utilized in various fields because of their excellent mechanical, thermal, chemical, optical properties, and good biocompatibility, especially in biomedical aspects. However, there is a concern that the unique characteristics of nanomaterials may have undesirable effects. Therefore, in this study, we sought to systematically investigate the effects of graphene quantum dots (GQDs) on the maturation of mouse oocytes and development of the offspring via in vitro and in vivo studies. In vitro, we found that the first polar body extrusion rate in the high dosage exposure groups (1.0-1.5 mg/ml) 2 decreased significantly and the failure of spindle migration and actin cap formation after GQDs exposure was observed. The underlying mechanisms might be associated with reactive oxygen species accumulation and DNA damage. Moreover, transmission electron microscope studies showed that GQDs may have been internalized into oocytes, tending to accumulate in the nucleus and severely affecting mitochondrial morphology, which included swollen and vacuolated mitochondria accompanied by cristae alteration with a lower amount of dense mitochondrial matrix. In vivo, when pregnant mice were exposed to GQDs at 8.5 days of gestation (GD, 8.5), we found that high dosage of GQD exposure (30 mg/kg) significantly affected mean fetal length; however, all the second generation of female mice grew up normal, attained sexual maturity, and gave birth to a healthy offspring after mating with a healthy male mouse. The results presented in this study are important for the future investigation of GQDs for the biomedical applications.

Di(n-butyl) phthalate exposure impairs meiotic competence and development of mouse oocyte.

Di(n-butyl) phthalate (DBP) is extensively used in industrial applications as plasticizer and stabilizer and its presence in the environment may present health risks for human. Previous studies have demonstrated its mutagenic, teratogenic, and carcinogenic ability. However, its effect on mammalian oocyte maturation remains unknown. In this study, we examined the effect of DBP on oocyte maturation both in vitro and in vivo. Our results showed that DBP could significantly reduce mice oocyte germinal vesicle breakdown (GVBD) and polar body extrusion (PBE) rates. In addition, oocyte cytoskeleton was damaged and cortical granule-free domains (CGFDs) were also disrupted. Finally, DBP induced early apoptosis of oocyte and granulosa cells (GCs). Collectively, these data demonstrate that DBP could reduce meiosis competence and mouse oocyte development.

A teleost TFPI-2 peptide that possesses a broad antibacterial spectrum and immune-stimulatory properties.

Tissue factor pathway inhibitor 2 (TFPI-2) is an analogue of TFPI-1 and a potent endogenous inhibitor of tissue factor (TF)-mediated blood coagulation. Previous reports have shown that several peptides derived from human and vertebrates TFPI-2 possess antibacterial activity against diverse bacteria. In this study, a C-terminal peptide, TO24 (with 24 amino acids), derived from red drum (Sciaenops ocellatus) TFPI-2, was synthesized and investigated for its antimicrobial spectrum, action mode, as well as the immune-stimulatory property. Our results indicated that TO24 was active against Gram-positive bacteria Micrococcus luteus and Staphylococcus aureus; Gram-negative bacteria Vibrio litoralis, Vibrio ichthyoenteri, Vibrio vulnificus and Vibrio scophthalmi, as well as fish megalocytivirus, infectious spleen and kidney necrosis virus (ISKNV). During its interaction with V. vulnificus, TO24 exerted its antibacterial activity by destroying cell membrane integrity, penetrating the cytoplasm and inducing degradation of genomic DNA and total RNA. In addition, TO24 had no hemolytic activity against red drum blood cells. In vitro, TO24 enhanced bactericidal activity of red drum macrophages. In vivo, administration of red drum with TO24 before bacterial infection significantly reduced pathogen dissemination and replication in tissues. These results indicate that TO24 is a broad-spectrum antimicrobial peptide with immune-stimulatory properties and it has the potential to be used as an antimicrobial agent in aquaculture.

TO17: A teleost antimicrobial peptide that induces degradation of bacterial nucleic acids and inhibits bacterial infection in red drum, Sciaenops ocellatus.

Tissue factor pathway inhibitor (TFPI)-1 is well known for its role as an inhibitor of blood coagulation. Several studies have demonstrated that the C-terminal peptides of TFPI-1 are active against a broad spectrum of microorganisms. In a previous study, we found that TO17 (with 17 amino acids), a TFPI-1 C-terminal peptide from red drum (Sciaenops ocellatus), was active against Edwardsiella tarda. In the present study, we investigated further the antimicrobial spectrum, action mode, as well as the immunostimulatory property of TO17. Our results showed that TO17 displayed antimicrobial activity against Staphylococcus aureus, Micrococcus luteus, Vibrio vulnificus, and infectious spleen and kidney necrosis virus, independent of host serum. Furthermore, the activity of TO17 was influenced by the length or type of amino acids at the N and C termini. During its interaction with V. vulnificus, TO17 exerted its antibacterial activity by destroying cell membrane integrity, penetrating the cytoplasm and inducing degradation of genomic DNA and total RNA. In addition, TO17 had no hemolytic activity against red drum blood cells. In vitro, TO17 enhanced production of nitric oxide and bactericidal activity of red drum macrophages. In vivo, administration of red drum with TO17 before bacterial infection significantly reduced pathogen dissemination and replication in tissues. These results indicate that TO17 is a broad-spectrum antimicrobial peptide with immunostimulatory properties and it has the potential to be used as an antimicrobial agent in aquaculture.

SKAP2 regulates Arp2/3 complex for actin-mediated asymmetric cytokinesis by interacting with WAVE2 in mouse oocytes.

SKAP2 (Src kinase-associated phosphoprotein 2), a substrate of Src family kinases, has been suggested to be involved in actin-mediated cellular processes. However, little is known about its role in mouse oocyte maturation. In this study, we thus investigated the expression, localization, and functions of SKAP2 during mouse oocyte asymmetric division. SKAP2 protein expression was detected at all developmental stages in mouse oocytes. Immunofluorescent staining showed that SKAP2 was mainly distributed at the cortex of the oocytes during maturation. Treatment with cytochalasin B in oocytes confirmed that SKAP2 was co-localized with actin. Depletion of SKAP2 by injection with specific short interfering RNA caused failure of spindle migration, polar body extrusion, and cytokinesis defects. Meanwhile, the staining of actin filaments at the oocyte membrane and in the cytoplasm was significantly reduced after these treatments. SKAP2 depletion also disrupted actin cap and cortical granule-free domain formation, and arrested a large proportion of oocytes at the telophase stage. Moreover, Arp2/3 complex and WAVE2 expression was decreased after the depletion of SKAP2 activity. Our results indicate that SKAP2 regulates the Arp2/3 complex and is essential for actin-mediated asymmetric cytokinesis by interacting with WAVE2 in mouse oocytes.

A TFPI-1 peptide that induces degradation of bacterial nucleic acids, and inhibits bacterial and viral infection in half-smooth tongue sole, Cynoglossus semilaevis.

Tissue factor pathway inhibitor 1 (TFPI-1) is a serine protease inhibitor that inhibits tissue factor (TF)-mediated coagulation. The C-terminal region of TFPI-1 could be cleaved off and proved to be antimicrobial against a broad-spectrum of microorganism. In a previous study, a C-terminal peptide, TC24 (with 24 amino acids), derived from tongue sole (Cynoglossus semilaevis) TFPI-1, was synthesized and found antibacterial against Micrococcus luteus. In the present study, the antibacterial spectrum and the action mode of TC24 was further examined, and its in vivo function was analyzed. Our results showed that TC24 also possesses bactericidal activity against Staphylococcus aureus and Vibrio vulnificus. During its interaction with the target bacterial cells, TC24 destroyed cell membrane integrity, penetrated into the cytoplasm, and induced degradation of genomic DNA and total RNA. In vivo study showed that administration of tongue sole with TC24 before bacterial and viral infection significantly reduced pathogen dissemination and replication in tissues. These results indicated that TC24 is a novel antimicrobial peptide against bacterial and viral pathogens, and that the observed effect of TC24 on bacterial RNA adds new insights to the action mechanism of fish antimicrobial peptides. Moreover, TC24 may play an important role in fighting pathogenic infection in aquaculture.

Protective effects of resveratrol against mancozeb induced apoptosis damage in mouse oocytes.

Mancozeb, a mixture of ethylene-bis-dithiocarbamate manganese and zinc salts, is one of the most widely used fungicides in agriculture. Mancozeb could lead to mitochondria dysfunction, cellular anti-oxidation enzymes depletion and apoptotic pathways activation. Previous studies indicated the exposure of mancozeb through mother would lead to irregular estrous cycles, decreased progesterone levels, reduced litter sizes, and more frequent delivery of dead fetuses. In this study, we investigated mancozeb inducing reproductive toxicity, especially focusing on its apoptotic effect and epigenetic modifications. We also showed that resveratrol, a kind of phytoalexin found in peanuts and grapes, can alleviate mancozeb's adverse effects, such as declined fertility, decreased ovary weight and primary follicles. Besides, mancozeb treated oocytes displayed suboptimal developmental competence and this can also be improved by treatment of resveratrol. More detailed investigation of these processes revealed that mancozeb increased reactive oxygen species, causing cell apoptosis and abnormal epigenetic modifications, and resveratrol can block these cytotoxic changes. Collectively, our results showed that resveratrol can alleviate mancozeb induced infertility and this was mainly through the correction of apoptotic tendency and the abnormity of cellular epigenetic modification.

KIF2A regulates the spindle assembly and the metaphase I-anaphase I transition in mouse oocyte.

KIF2A, a member of the kinesin-13 family, has been reported to play a role in spindle assembly in mitosis. However, its function in mammalian meiosis remains unknown. In this research, we examined the expression, localization and function of KIF2A during mouse oocyte meiosis. KIF2A was expressed in some key stages in mouse oocyte meiosis. Immunofluorescent staining showed that KIF2A distributed in the germinal vesicle at the germinal vesicle stage and as the spindle assembling after meiosis resumption, KIF2A gradually accumulated to the entire spindle. The treatment of oocytes with taxol and nocodazole demonstrated that KIF2A was co-localized with α-tubulin. Depletion of KIF2A by specific short interfering (si) RNA injection resulted in abnormal spindle assembly, failure of spindle migration, misaligned chromosomes and asymmetric cell division. Meanwhile, SKA1 expression level was decreased and the TACC3 localization was disrupted. Moreover, depletion of KIF2A disrupted the actin cap formation, arrested oocytes at metaphase I with spindle assembly checkpoint protein BubR1 activated and finally reduced the rate of the first polar body extrusion. Our data indicate that KIF2A regulates the spindle assembly, asymmetric cytokinesis and the metaphase I-anaphase I transition in mouse oocyte.