理学最新文献

Backgroud: Efficient communication between the embryo and the endometrium is essential for the successful establishment and maintenance of pregnancy. Uterine-derived extracellular vesicles (EVs) contribute to embryo-maternal communication, supporting early embryonic development. This study aimed to: (i) compare the protein cargo of uterine fluid EVs (UF-EVs) from CYCLIC and PREGNANT heifers; (ii) characterize the protein profile of conditioned medium (CM)-EVs from endometrial explants cultured alone (EXPL) or co-cultured with five d 7 blastocysts (EXPL + EMB) in vitro; and (iii) compare the EV protein cargo between the in vivo and in vitro models (i.e., EXPL vs. CYCLIC and EXPL + EMB vs. PREGNANT).

Results: We identified 1,459 and 1,752 proteins in the UF-EVs of CYCLIC and PREGNANT heifers, respectively. Among these, 12 were exclusive to CYCLIC, and 18 were exclusive to PREGNANT. Among the 1,329 proteins identified in both groups, 16 were differently abundant; ten were more abundant, and six were less abundant in UF-EVs from PREGNANT heifers. In vivo, the changes in UF-EV protein cargo induced by the presence of a blastocyst were related to inflammatory and immune responses, endometrial receptivity, and support of early embryonic development by promoting cell polarity, cell-cell adhesion, and stem cell differentiation. In vitro, we identified 1,501 proteins in the CM-EVs from EXPL, 1,975 in the CM-EVs from EXPL + EMB, and 82 in the CM-EVs from EMB. Additionally, 50 proteins were unique to EXPL + EMB, and another 33 were differentially abundant due to the synergistic interaction between the embryo and the endometrium. These proteins are involved in embryonic development, regulation of stem cell differentiation, establishment and maintenance of cell polarity, interferon tau (IFNT)-mediated cell signaling, endometrial receptivity, and immune modulation. Although there are qualitative and quantitative differences between in vivo and in vitro-derived EVs, UF-EVs from CYCLIC heifers compared to CM-EVs from EXPL, as well as UF-EVs from PREGNANT heifers compared to CM-EVs from EXPL + EMB shared common proteins.

Conclusions: These findings highlight the pivotal role of EVs in embryo-maternal communication, suggesting that their protein cargo may actively contribute to the modulation of the uterine environment to support early embryonic development. Understanding these molecular interactions could provide valuable insights into the mechanisms of implantation and pregnancy establishment.

Background: Mastitis is known to alter milk lipid yield, but its effects on lipid composition in blood and milk remain less understood. This study investigated changes in fatty acid (FA) composition in blood lipid fractions and milk of dairy cows following an intramammary lipopolysaccharide (LPS) challenge and explored potential links associated with these changes. We hypothesized that intramammary LPS infusion would alter the FA composition of blood lipid fractions, and that milk FA composition would reflect these changes. Furthermore, we hypothesized that prostaglandin E₂ (PGE₂) would be associated with changes in both blood and milk FA composition, functioning as a potential mediator of these changes.

Results: Ten lactating cows were split into two groups. The treatment group received intramammary infusions of 50 μg Escherichia coli LPS in both quarters of one udder half to induce clinical mastitis, and saline infusions in the quarters of the opposite udder half; the control group received saline infusions in one udder half only. Blood and foremilk were collected from individual cows or glands at -1, 3, 6, 12, and 24 h post-infusion. Blood lipids were fractionated into cholesterol esters, free fatty acids (FFA), phospholipids (PL), and triacylglycerides (TAG). The FA composition was analyzed via gas-liquid chromatography. Total plasma TAG, FFA, and PGE₂ concentrations were measured by colorimetric assay or ELISA. Statistical significance was determined using mixed models with Tukey's test. Lipopolysaccharide infusion did not affect total plasma TAG and FFA concentrations but increased plasma PGE₂ concentrations and Δ9 desaturation indices in plasma TAG. A distinct shift in FA composition in plasma phospholipids and TAG was observed between the treatment and control groups at 6 and 12 h post-infusion. Specifically, LPS increased the proportion of n-6 polyunsaturated FA (18:2, 18:3, 20:3, 20:4, 20:5) and FA with less than 16 carbons while decreasing the saturated FA (18:0 and 20:0) in plasma TAG at 6 and 12 h. However, the milk FA composition remained unchanged.

Conclusion: Our findings indicate that transient intramammary LPS challenge influences systemic lipid metabolism without altering the milk FA composition, suggesting that mammary inflammatory responses affect blood lipids independently of milk lipid secretion.

Background: Follicular atresia, a complex degenerative process regulated by multiple molecular mechanisms, significantly affects female reproductive performance in animals. While granulosa cell (GC) apoptosis has been well established as a primary mechanism underlying follicular atresia, the potential involvement of ferroptosis, which is an iron-dependent form of regulated cell death, remains largely unexplored in chickens.

Results: Using a tamoxifen (TMX)-induced avian model of follicular atresia, we demonstrated that ferroptosis plays a critical role in follicular degeneration. Inhibition of ferroptosis through pharmacological agents significantly restored follicular function, underscoring its potential as a therapeutic target. Notably, we observed a significant upregulation of ubiquitin-specific peptidase 9, X-linked (USP9X) in GCs during atresia. Through comprehensive in vitro and in vivo investigations, we confirmed that USP9X facilitates follicular atresia by promoting ferroptosis in GCs. Mechanistically, USP9X induces ferroptosis by stabilizing Beclin1 through deubiquitination, thereby activating autophagy-dependent ferroptosis. This pathway was effectively suppressed by autophagy inhibitors, emphasizing the essential role of autophagy in USP9X-mediated ferroptosis.

Conclusions: Our findings provide the evidence that the USP9X-Beclin1 axis regulates autophagy-dependent ferroptosis during avian follicular atresia. These insights reveal novel molecular targets and potential genetic markers for improving reproductive efficiency in chicken breeding programs.

Background: To more effectively address the scarcity resources and elevated costs associated with fishmeal (FM), the utilization of cottonseed protein concentrate (CPC) as an alternative in aquaculture feeds has become increasingly prevalent. However, high levels of CPC substitution for FM have been reported to suppress the growth of fish and impair intestinal health. Hydrolysable tannin (HT) has been reported to exhibit biological activities such as anti-inflammatory and antioxidant activities, but whether the HT can generate positive biological effects on the intestinal health of largemouth bass (Micropterus salmoides) remains unknown. Largemouth bass (initial weight: 6.03 ± 0.01 g) were subjected to an 8-week feeding trial with three different diets: a basic diet (named as the NC), a high CPC diet (in which CPC replaced 75% of the FM protein in the NC diet, named as the HC), and an additive diet (1.25 g/kg of the HT was added to the HC diet, named as the HCH) to explore the potential benefits of HT on intestinal health.

Results: The HC treatment significantly reduced the weight gain rate of fish, increased the feed conversion ratio, and induced intestinal inflammation. However, the HCH treatment could alleviate the adverse impacts of the HC diet, as evidenced by the promotion of growth and feed utilization, increased activity of digestive enzymes and antioxidant capacities, downregulated expression of pro-inflammatory factors, and maintenance of the integrity of intestinal barrier. Metabolomic analysis revealed that HCH treatment could reduce the pro-inflammatory active substances produced by arachidonic acid metabolism, including prostaglandin F_2α (PGF_2α) and leukotriene B₄ (LTB₄). Transcriptomic results indicated that dietary HT might alleviate intestinal inflammation by suppressing the activation of the NF-κB signaling pathway. Furthermore, correlation analysis demonstrated that the metabolites PGF_2α and LTB₄, derived from arachidonic acid, exhibited a significant positive correlation with the expression of genes associated with pro-inflammatory responses within the NF-κB signaling pathway.

Conclusions: The study indicates that the HT mitigates the growth retardation and intestinal inflammation resulting from the HC diet on largemouth bass.

Background: Backfat thickness (BFT) is a vital economic trait in pigs, reflecting subcutaneous fat levels that affect meat quality and production efficiency. As a complex trait shaped by multiple genetic factors, BFT has been studied using genome-wide association studies (GWAS) and linkage analyses to locate fat-related quantitative trait loci (QTLs), but pinpointing causal variants and genes is hindered by linkage disequilibrium and limited regulatory data. This study aimed to dissect the QTLs affecting BFT on Sus scrofa chromosome 1 (SSC1), elucidating regulatory variants, effector genes, and the cell types involved.

Results: Using whole-genome genotyping data from 3,578 pigs and phenotypic data for five BFT traits, we identified a 630.6 kb QTL on SSC1 significantly associated with these traits via GWAS and fine-mapping, pinpointing 34 candidate causal variants. Using deep convolutional neural networks to predict regulatory activity from sequence data integrated with detailed pig epigenetic profiles, we identified five SNPs potentially affecting enhancer activity in specific tissues. Notably, rs342950505 (SSC1:161,123,588) influences weak enhancer activity across multiple tissues, including the brain. High-throughput chromosome conformation capture (Hi-C) analysis identified that rs342950505 interacts with eight genes. Chromatin state annotations confirmed enhancer activity at this QTL in the cerebellum. Leveraging these insights, single-cell ATAC-seq revealed a chromatin accessibility peak encompassing rs342950505 that regulates PMAIP1 expression in inhibitory neurons via enhancer-mediated mechanisms, with an adjacent peak modulating CCBE1 expression in neuroblasts and granule cells. Transcriptome-wide association studies (TWAS) confirmed PMAIP1's role in the hypothalamus, and Mendelian randomization (MR) validated PMAIP1 and CCBE1 as key brain expression quantitative trait locus (eQTL) effectors. We propose that the variant rs342950505, located within a regulatory peak, modulates PMAIP1 expression in inhibitory neurons, potentially influencing energy homeostasis via hypothalamic regulation. Similarly, CCBE1 may contribute to this process.

Conclusions: Our results, through systematic dissection of pleiotropic BFT-associated loci, provide a framework to elucidate regulatory mechanisms of complex traits, offering insights into polygenic control through lipid metabolism and neural signaling pathways.