Fidgetin knockdown and knockout influences female reproduction distinctly in mice

Microtubule-severing proteins(MTSPs),are a family of proteins which use adenosine triphosphate to sever microtubules.MTSPs have been shown to play an important role in multiple microtubule-involved cellular processes.One member of this family,fidgetin(FIGN),is also involved in male fertility;however,no studies have explored its roles in female fertility.In this study,we found mouse fidgetin is rich within oocyte zona pellucida(ZP)and is the only MTSP member to do so.Fidgetin also appears to interact with all three ZP proteins.These findings prompted us to propose that fidgetin might prevent polyspermy.Results from in vitro maturation oocytes analysis showed that fidgetin knockdown did cause polyspermy.We then deleted all three fidgetin isoforms with CRISPR/Cas9 technologies;however,female mice remained healthy and with normal fertility.Of all mouse MTSPs,only the mRNA level of fidgetin-like 1(FIGNL1)significantly increased.Therefore,we assert that fidgetin-like 1 compensates fidgetin's roles in fidgetin knockout female mice.

Effects of nuciferine on Nrf2/HO-1 signaling pathway in adipose tissue of obesity model rats

Objective:This study aimed to explore the therapeutic effect of nuciferine on high-fat diet-induced obesity in rats and the influence of nuciferine on nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway in the adipose tissue.Methods:A total of 40 male Sprague Dawley(SD)rats were evenly divided into the normal,model,positive control,and nuciferine groups,using the random number table method.Except for the normal group,rats in the other groups were fed with high-fat diet for 12 weeks to establish the obesity model.During the model establishment,rats in the positive control group received atorvastatin calcium 2 mg/kg,rats in the nuciferine group received nuciferine 20 mg/kg,and rats in the normal and model groups received normal saline 2 mL,daily through intragastric administration for 12 consecutive weeks.After model establishment and administration,the body weight,Lee’s index,and blood lipids of rats in each group were measured,and hematoxylin and eosin(HE)staining was performed on the liver and adipose tissues to evaluate the therapeutic effect of nuciferine on obesity rat model.Additionally,the levels of superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)in the serum of rats in each group were determined,and the gene expressions of Nrf2 and HO-1 in the adipose tissue of rats in each group were detected through quantitative polymerase chain reaction(qPCR)to investigate the mechanism of action of nuciferine in the treatment of obesity.Results:After 12 weeks of model establishment and administration,we observed that compared with the model group,nuciferine could significantly reduce the body weight,Lee’s index,and serum triglyceride(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C)levels and increase the serum high-density lipoprotein cholesterol(HDL-C)level in obesity rat model(P<0.05 or P<0.01).HE staining revealed that nuciferine could significantly alleviate liver steatosis in obesity rat model and improve the cell morphology in epididymal adipose tissue.Moreover,nuciferine could elevate serum SOD and GSH-Px activities in obesity rat model and lower the serum MDA level(P<0.05 or P<0.01).The qPCR indicated that nuciferine could upregulate the gene expression of Nrf2 and HO-1 in the adipose tissue of obesity rat model(P<0.05 or P<0.01).

ATP50 Hypo-crotonylation Caused by HDAC2 Hyper-Phosphorylation Is a Primary Detrimental Factor for Downregulated Phospholipid Metabolism under Chronic Stress

Objective.Chronic stress(CS)-induced abnormal metabolism and other subsequent aspects of abnormality are threatening human health.Little is known regarding whether and how protein post-translational-modifications(PTMs)correlate with abnormal metabolism under CS.The aim of this study was to address this issue and also identify novel key protein PTM.Methods.First,we screened which pan-PTM had significant change between control and CS female mice and whether clinical CS females had similar pan-PTM change.Second,we performed quantitative PTM-omics and metabolomics to verify the correlation between abnormal protein PTMs and atypical metabolism.Third,we performed quantitative phospho-omics to identify the key PTM-regulating enzyme and investigate the interaction between PTM protein and PTM-regulating enzyme.Fourth,we attempted to rectify the abnormal metabolism by correcting the activity of the PTM-regulating enzyme.Finally,we examined whether the selected key protein was also correlated with stress scores and atypical metabolism in clinical women.Results.We initially found that multiple tissues of CS female mice have downregulated pan-crotonylation,and verified that the plasma of clinical CS females also had downregulated pan-crotonylation.Then we determined that ATP5O-K51 crotonylation decreased the most and also caused gross ATP5O decrement,whereas the plasma of CS mice had downregulated phospholipids.Next,downregulating ATP5O crotonylation partially recapitulated the downregulated phospholipid metabolism in CS mice.Next,we verified that HDAC2-S424 phosphorylation determined its decrotonylation activity on ATP5O-K51.Furthermore,correcting HDAC2 hyper-phosphorylation recovered the gross ATP5O level and partially rescued the downregulated phospholipid metabolism in CS mice.Finally,the ATP5O level was also significantly iower and correlated with high stress scores and downregulated phospholipid metabolism in clinical female plasma.Conclusion.This study discovered a novel PTM mechanism involving two distinct types of PTM in CS and provided a novel reference for the clinical precautions and treatments of CS.