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Abstract Background 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. Results 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. Conclusions 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.

Canine endometritis is a common disease in dogs. This work intends to establish the model of lipopolysaccharide (LPS) -induced canine endometritis, and investigate the effect of Tanshinone IIA (Tan IIA) on canine endometritis. At first, we tested the rectal temperature and the production of IL-1β and TNF-α at 6 h, 12 h, 18 h and 24 h after LPS administration. Then 9 beagles were divided into 3 groups on average, all beagles received intraperitoneal injection of saline solution (group 1 and 2) or Tan IIA (group 3) at 6 h before and after LPS challenge. Beagles of group 2 and 3 were performed uterine infusion of LPS, and beagles of group 1 were performed uterine infusion of saline solution. The rectal temperature was measured 6 h, 12 h, 18 h, 24 h post-LPS challenge, all uterus were collected after 24 h post-LPS challenge. The results showed that canine endometritis can be established by LPS at the concentration of 0.5 mg/kg of body weight after 24 h performance. The rectal temperature, the production of IL-1β and TNF-α increased significantly when the model was established. The results showed that rectal temperature, production of IL-1β and TNF-α and the expression of IL-6 were significantly reduced after treatment with Tan IIA compared with the group of LPS challenge only. However, the expression of IL-10 increased after Tan IIA treatment. Considering the positive anti-inflammation effect on the LPS-induced canine endometritis, Tan IIA may be used as a therapeutic agent to treat the clinical canine endometritis.

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.

Objectives Clinical endometritis is a common reproductive disorder in mammals that seriously endangers animal health and causes economic losses worldwide. This study aims to use lipopolysaccharide and Trueperella pyogenes exotoxin as modelling reagents (LC) to perfuse the mouse uterus in order to establish a model of clinical endometritis and to investigate the anti-inflammatory and antioxidant effects of chlorogenic acid (CGA). Methods In this study, five LC uterine perfusions were selected to model clinical endometritis. The anti-inflammatory and antioxidant effects of CGA were clarified. Through HE staining, proinflammatory cytokines, blood testing, NFκB and Keap1/Nrf2 signalling pathways and other index changes to explore the protection mechanism of CGA. Key findings After CGA treatment, the appearance, inflammatory damage and blood indicators of the mouse uterus returned to normal. Simultaneously, CGA could inhibit the activation of NFκB and reduce the release of inflammatory cytokines; CGA could also activate Keap1/Nrf2, promote the dissociation of Keap1 and Nrf2 and significantly increase the expression of the downstream genes HO-1 and NQO1. Conclusions The above results together explain that five LC uterine perfusions can be used to establish a mouse model of clinical endometritis. CGA can treat clinical endometritis by activating Keap1/Nrf2 and inhibiting the NFκB signalling pathway. [ABSTRACT FROM AUTHOR]

Background. The DNAJ family of molecular chaperones maintains protein homeostasis in mitotic and postmeiotic cells, especially germ cells. Recently, we found that the transcription factor SOX30 initiates transcription of Dnajb8 during late meiosis and spermiogenesis in mouse testes. Methods. We used the CRISPR/Cas9 system to generate Dnajb8 mutant mice and analyze the phenotype of the Dnajb8 mutants. Results. Although Dnajb8 is an evolutionarily conserved gene, it is not essential for spermatogenesis and male fertility. We provide this phenotypic information, which could prevent duplicative work by other groups. [ABSTRACT FROM AUTHOR]