Transcription factors of the Sox protein family contain a DNA-binding HMG box and are key regulators of progenitor cell fate. Here, we report that expression of Sox30 is restricted to meiotic ...spermatocytes and postmeiotic haploids.
mutant males are sterile owing to spermiogenic arrest at the early round spermatid stage. Specifically, in the absence of Sox30, proacrosomic vesicles fail to form a single acrosomal organelle, and spermatids arrest at step 2-3. Although most
mutant spermatocytes progress through meiosis, accumulation of diplotene spermatocytes indicates a delayed or impaired transition from meiotic to postmeiotic stages. Transcriptome analysis of isolated stage-specific spermatogenic cells reveals that Sox30 controls a core postmeiotic gene expression program that initiates as early as the late meiotic cell stage. ChIP-seq analysis shows that Sox30 binds to specific DNA sequences in mouse testes, and its genomic occupancy correlates positively with expression of many postmeiotic genes including
,
,
and
These results define Sox30 as a crucial transcription factor that controls the transition from a late meiotic to a postmeiotic gene expression program and subsequent round spermatid development.
Berberine exerts many beneficial effects on lipopolysaccharide (LPS)‐induced bovine endometrial epithelial cells (BEECs). Recently, we also found that berberine shows significant antiapoptotic and ...autophagy‐promoting activities, but the underlying mechanism has not been elucidated. This research explored the association between the antiapoptotic and autophagy‐promoting activities of berberine in LPS‐treated BEECs. BEECs were first preconditioned with an inhibitor of autophagic flux (chloroquine CQ) for 1 h, treated with berberine for 2 h, and then incubated with LPS for 3 h. Cell apoptosis was assessed by flow cytometry, and autophagy activities were assessed by immunoblot analysis of LC3II and p62. The results indicated that the antiapoptotic activity of berberine was notably inhibited in LPS‐treated BEECs after preconditioning with CQ for 1 h. Furthermore, to determine whether berberine promoted autophagy by activating the nuclear factor‐erythroid 2 related factor 2 (Nrf2) signaling pathway, we assessed autophagy in LPS‐treated BEECs after preconditioning with a signaling pathway inhibitor of Nrf2 (ML385). The results indicated that the enhanced autophagy activity induced by berberine was partially reversed in LPS‐treated BEECs after the Nrf2 signaling pathway was disturbed by ML385. In conclusion, berberine enhances autophagic flux to allow resistance to LPS‐induced apoptosis by activating the Nrf2 signaling pathway in BEECs. The present study may provide new insight into the antiapoptotic mechanism of berberine in LPS‐induced BEECs.
ABSTRACT
Rapamycin is a clinically important drug that is used in transplantation and cancer therapy but which causes a number of side effects, including male infertility. Its canonical target, ...mammalian target of rapamycin complex 1 (mTORC1), plays a key role in metabolism and binds chromatin; however, its precise role in the male germline has not been elucidated. Here, we inactivate the core component, Raptor, to show that mTORC1 function is critical for male meiosis and the inactivation of sex chromosomes. Disruption of the Raptor gene impairs chromosomal synapsis and prevents the efficient spreading of silencing factors into the XY chromatin. Accordingly, mRNA for XY‐linked genes remains inappropriately expressed in Raptor‐deficient mice. Molecularly, the failure to suppress gene expression corresponded with deficiencies in 2 repressive chromatin markers, H3K9 dimethylation and H3K9 trimethylation, in the XY body. Together, these results demonstrate that mTORC1 has an essential role in the meiotic progression and silencing of sex chromosomes in the male germline, which may explain the infertility that has been associated with such inhibitors as rapamycin.—Xiong, M., Zhu, Z., Tian, S., Zhu, R., Bai, S., Fu, K., Davis, J. G., Sun, Z., Baur, J. A., Zheng, K., Ye, L. Conditional ablation of Raptor in the male germline causes infertility due to meiotic arrest and impaired inactivation of sex chromosomes. FASEB J. 31, 3934–3949 (2017). www.fasebj.org—Xiong, Mengneng, Zhu, Zhiping, Tian, Suwen, Zhu, Ruping, Bai, Shun, Fu, Kaiqiang, Davis, James G., Sun, Zheng, Baur, Joseph A., Zheng, Ke, Ye, Lan, Conditional ablation of Raptor in the male germline causes infertility due to meiotic arrest and impaired inactivation of sex chromosomes. FASEB J. 31, 3934–3949 (2017)
RNA regulation by RNA-binding proteins (RBPs) involve extremely complicated mechanisms. MOV10 and MOV10L1 are two homologous RNA helicases implicated in distinct intracellular pathways. MOV10L1 ...participates specifically in Piwi-interacting RNA (piRNA) biogenesis and protects mouse male fertility. In contrast, the functional complexity of MOV10 remains incompletely understood, and its role in the mammalian germline is unknown. Here, we report a study of the biological and molecular functions of the RNA helicase MOV10 in mammalian male germ cells.
MOV10 is a nucleocytoplasmic protein mainly expressed in spermatogonia. Knockdown and transplantation experiments show that MOV10 deficiency has a negative effect on spermatogonial progenitor cells (SPCs), limiting proliferation and in vivo repopulation capacity. This effect is concurrent with a global disturbance of RNA homeostasis and downregulation of factors critical for SPC proliferation and/or self-renewal. Unexpectedly, microRNA (miRNA) biogenesis is impaired due partially to decrease of miRNA primary transcript levels and/or retention of miRNA via splicing control. Genome-wide analysis of RNA targetome reveals that MOV10 binds preferentially to mRNAs with long 3'-UTR and also interacts with various non-coding RNA species including those in the nucleus. Intriguingly, nuclear MOV10 associates with an array of splicing factors, particularly with SRSF1, and its intronic binding sites tend to reside in proximity to splice sites.
These data expand the landscape of MOV10 function and highlight a previously unidentified role initiated from the nucleus, suggesting that MOV10 is a versatile RBP involved in a broader RNA regulatory network.
Kaempferol isolated from the root of
Zingiberaceae plants
galangal and other Chinese herbal medicines have been reported to have anti-inflammatory properties. However, the anti-inflammatory effects ...of kaempferol on lipopolysaccharide (LPS)-induced mastitis are unknown and their underlying molecular mechanisms remain to be explored. The aim of this study was to evaluate the effects of kaempferol on LPS-induced mouse mastitis. The mouse model of mastitis was induced by injection of LPS through the duct of mammary gland. Kaempferol was injected 1 h before and 12 h after induction of LPS intraperitoneally. The present results showed that kaempferol markedly reduced infiltration of neutrophilic granulocyte, activation of myeloperoxidase (MPO), expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in a dose-dependent manner, which were increased in LPS-induced mouse mastitis. Furthermore, kaempferol suppressed the phosphorylation of nuclear factor-κB (NF-κB) p65 subunit and the degradation of its inhibitor IκBα. All results suggest that anti-inflammatory effects of kaempferol against the LPS-induced mastitis possibly through inhibition of the NF-κB signaling pathway. Kaempferol may be a potential therapeutic agent for mastitis.
Metritis is a frequently occurring diseases in postpartum cows and is one of the important reasons for the infertility of dairy cows, accounting for 20–30% of dairy cow diseases and has serious ...implications for the dairy industry. It has been reported in the literature that the bacterial balance of genital tracts is directly related to the maintenance of physiological function and the development of various diseases of the reproductive system. By analyzing the changes in abundance and diversity of bacteria in the cow uterus from 1 to 35 days postpartum, the objective was to reveal the mechanism of metritis in cows and provide the basis for diagnosis, treatment and prevention of metritis in postpartum dairy cows. Uterine contents were taken from six cows (three healthy and three with metritis) on 1, 7, 14, 21 and 35 days after parturition. DNA genomes extracted from the samples were primed with 515F5′-GTGCCAGCMGCCGCGG-3′ and 907R5′-CCGTCAATTCMTTRAGTTT-3′ for PCR amplification of the V4+V5 regions of the 16S rDNA genes and construction of a gene library. The sequence of the bacterial structure of the cow uterine contents was analyzed using 16S rDNA high-throughput sequencing technology.
A total of 30 samples were tested by PCR, and 29 samples qualified. The results of cluster analysis showed that except for one sample, the number of OTUs in the healthy cows was above 200, while in the cows with metritis, except for three samples, OTUs were below 200. The Chao1 and Shannon indices showed that the abundance of bacteria in the cow uterus was lower than that of healthy cows.
Analysis of the relative abundance of bacteria in the cow uterus showed that there were six phyla present, including Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Actinobacteria and Tenericutes. There were 10 dominant genera in healthy cows, including Bacteroides, Clostridium sensu stricto 1, Escherichia-Shigella, Fusobacterium, Halomonas, Helcococcus, Porphyromonas, Prevotella 6, Rikenellaceae RC9 gut group and Streptococcus. There were nine dominant genera in cows with metritis, including Bacteroides, Caviibacter, Clostridium sensu stricto 1, Falsiporphyromonas, Fusobacterium, Halomonas, Helcococcus, Porphyromonas and Prevotella 7.
Phylogenetic tree analysis showed that uterine contents from 29 samples could be separated into two clusters. Eleven samples from the cows with metritis were clustered with one sample from the healthy group, and 13 samples from the healthy cows were clustered together with four samples from the metritis group. Principal co-ordinate analysis showed that the points representing healthy cows and those representing the metritis group were concentrated in two distinct regions, which shows that there were significant differences in the structure evolution between healthy cows and cows with metritis.
The above results indicate that bacterial diversity declines with time postpartum in healthy cows and is lower in cows with metritis, with characteristic changes in the relative abundances, including increases in Bacteroidetes and Fusobacteria, decreases in Firmicutes and Proteobacteria, increases in Porphyromonas, Bacteroides and Fusobacterium, and a decrease in Clostridium sensu stricto 1.
•Berberine significantly reduced the levels of CRP, IL-1β, IL-6, and TNF-α.•Berberine exhibited anti-inflammatory effect via activating Nrf2 signaling pathway.•Berberine regulated many signaling ...pathways, including the Nrf2 and NF-κB.
Endometritis is a common inflammatory disease that disturbs the rapid development of dairy farming. In the present study, we investigated the anti-inflammatory effects of berberine on the LPS-induced inflammatory response in bovine endometrial epithelial cells (bEECs) and the participation of the Keap1/Nrf2 signaling pathway in this process. Berberine treatment significantly reduced the LPS-induced expression levels of CRP, IL-1β, IL-6, and TNF-α in bEECs. The Nrf2 signaling pathway in these cells was also activated by berberine. We further evaluated the effects of Nrf2 activators and inhibitors on the downregulation of proinflammatory cytokines. The activator of Nrf2 significantly inhibited the production of these proinflammatory cytokines that was induced by LPS. However, an inhibitor of Nrf2 only partially inhibited the anti-inflammatory effects of berberine on the LPS-induced inflammatory response in bEECs. In conclusion, our findings suggest that berberine exerts anti-inflammatory effects partially by activating the Keap1/Nrf2 signaling pathway.
Mastitis is the most prevalent disease in dairy cows worldwide. Evidence has emerged that oxidative stress plays a crucial role in the development of mastitis. This study aimed to investigate the ...antioxidative effects of tanshinone IIa (Tan IIa) on LPS-induced oxidative stress in dairy cow mammary epithelial cells (CMECs).
We examined the levels of ROS and MDA in LPS-treated CMECs after supplementation with Tan IIa using detection kits and found that Tan IIa significantly inhibited the upregulation of these factors. In addition, we also found that Tan IIa significantly reversed the decrease in mitochondrial membrane potential induced by LPS. Moreover, Tan IIa improved the activities of antioxidant enzymes, which were decreased by LPS. Finally, we examined the probable pathway in which Tan IIa exerted its antioxidant effects using qPCR and western blotting and found that Tan IIa significantly activated the Keap1/Nrf2 signalling pathway.
These results suggest that Tan IIa might become a possible therapeutic agent for the treatment of dairy cow mastitis by weakening oxidative stress induced by LPS in CMECs.
•Tan IIa significantly reduced the level of ROS and MDA induced by LPS.•Tan IIa significantly reversed the decrease of mitochondrial membrane potential induced by LPS.•Tan IIa improving the activities of antioxidant enzymes which were decreased by LPS.•Tan IIa exerts anti-oxidative effects through activating the Keap1/Nrf2 signalling pathways.
Endometritis is one of the main causes of bovine infertility, which causes serious economic losses to the industry. The endometrium is the first line of defense against invading microbial pathogens ...in the uterus. Andrographolide is the primary active component of A. paniculate, and has been shown to have anti-inflammatory and antioxidant effects. However, its effects on the LPS-induced signaling pathway in bovine endometrial epithelial cells (bEECs) have not been reported yet. The aim of this study was to investigate the anti-inflammatory effects and mechanism of andrographolide in the LPS-induced inflammatory response of bEECs. We found that andrographolide strongly reduced LPS-induced NO and iNOS expression. The production of cytokines that were upregulated by LPS was significantly suppressed. To investigate the anti-inflammatory mechanism of andrographolide, we examined the activation of Nrf2. The results shown that andrographolide inhibited the expression of Keap1 but increased the expression of Nrf2. The expression levels of target genes of Nrf2 including Ho-1 and Nqo-1 were increased by andrographolide. Taken together, these results suggest that andrographolide may serve as a candidate to protect against the LPS-induced inflammatory response by inducing Nrf2 activation.
•Andrographolide remarkably reduced LPS-induced NO and iNOS expression;•Andrographolide reduced the expression level of IL-1β and IL-6 induced by LPS;•Andrographolide inhibited the expression of Keap1 but increased the expression of Nrf2;•The expression levels of target genes of Nrf2 including Ho-1 and Nqo-1 were increased by andrographolide;•Andrographolide exhibited anti-inflammatory effect via activating Nrf2 signaling pathway.
Mixed infection with Escherichia coli and Trueperella pyogenes (T. pyogenes) leads to purulent endometritis, but the underlying molecular mechanisms remain unclear. The aim of this study was to ...investigate the effect of tanshinone ⅡA (Tan ⅡA) on E. coli and T. pyogenes -induced purulent endometritis and explore the underlying mechanism. First, lipopolysaccharide (LPS) isolated from E. coli and bacteria-free filtrates (BFFs) isolated from T. pyogenes were used to induce a model of bovine endometrial epithelial cell (bEEC) damage in vitro. bEECs were pretreated with or without Tan ⅡA for 2 h, before LPS and BFFs were introduced to induce damage to investigate the protective effect of Tan IIA. Then, the cytolytic activity and inflammatory response in bEECs were examined using CCK-8, LDH and RT-qPCR assays. Furthermore, we confirmed the molecular mechanism by which Tan ⅡA reversed the damaged phenotypes in LPS- and BFFs-induced bEECs via the NF-κB/Snail2 pathway using qPCR and Western blotting. Tan ⅡA significantly decreased the cytolytic activity and inflammatory response in LPS- and BFFs-induced bEECs. In addition, Tan ⅡA reversed the dysregulation of E-cadherin, N-cadherin and vimentin. Moreover, Tan ⅡA significantly inhibited the activation of the NF-κB signaling pathway and decreased the expression level of Snail2, which is the main regulator of the epithelial-mesenchymal transition (EMT). In summary, Tan ⅡA inhibits the LPS-induced EMT and protects bEECs from pyolysin-induced damage by modulating the NF-κB/Snail2 signaling pathway.
•Tan ⅡA significantly decreased the cytolytic activity and the inflammatory response in bEECs induced by LPS and BFF.•Tan ⅡA reversed the LPS-induced downregulation of E-cadherin and upregulations of N-cadherin and vimentin.•Tan ⅡA repressed the migration of bEECs and the expression of inflammatory cytokines induced by LPS.•Tan ⅡA significantly inhibited the activation of NF-κB signaling pathway, and decreased the expression level of snail2.
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