FOXO1 x MAPK

Leiocassis longirostris (L. longirostris) is a significant freshwater fish species in China. Due to the decline in natural populations, its market demand is now largely met through artificial breeding. However, the complex and prolonged ovarian maturation process has rendered current artificial breeding methods limited. Additionally, the molecular mechanisms underlying ovarian development in L. longirostris remain poorly understood. In this study, we conducted RNA and protein sequencing of L. longirostris ovarian samples across five developmental stages, categorized based on histological staining results. To uncover the dynamic characteristics of ovarian development, we identified differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) by comparing gene and protein expression profiles stage-by-stage (stage I to II, II to III, III to IV, and IV to V). GO and KEGG enrichment analyses of these profiles revealed significant enrichment in terms and pathways related to the endocrine system, signal transduction, and lipid metabolism, with dynamic changes observed across stages. The nine-quadrant analysis further elucidated the regulatory characteristics of DEGs and DEPs, highlighting differences in mRNA and protein-level regulation of genes and proteins involved in fat and protein digestion and absorption. Protein-protein interaction networks were further constructed from shared DEGs and DEPs profiles to evacuate candidate proteins involved in L. longirostris ovarian development. Moreover, we clustered DEGs and DEPs into ten clusters based on their expression trend. KEGG enrichment analysis of clusters showed pathways such as FoxO and MAPK signaling pathways were significantly enriched in cluster 4, where the expression increased as ovarian development. qPCR validation confirmed differential expression of genes associated with oocyte meiosis and steroid hormone biosynthesis, consistent with RNA-seq results. This study provides a foundation for understanding yolk lipids metabolism, paracrine factors, and endocrine system regulation during ovarian development and offers insights into developing efficient artificial breeding strategies for L. longirostris.

The oviduct and uterus provide a supportive environment for early embryonic development, where effective embryo-maternal communication is essential for pregnancy success. Extracellular vesicles (EVs) from oviductal (OF) and uterine fluids (UF) are key mediators of this communication, transporting bioactive molecules such as proteins, lipids, DNA, and regulatory RNAs. Among these, microRNAs (miRNAs) are particularly interesting due to their role in modulating reproductive processes. Although underrepresented in EVs research, the rabbit offers a robust model owing to its inducible ovulation, controlled reproductive physiology, and similarities to human implantation dynamics. This study aims to uncover, in does, how EVs could influence key reproductive processes by isolating, characterizing, and analyzing the miRNA cargo from OF- and UF-EVs. We identified 388 miRNAs in both OF-EVs and UF-EVs, 18 miRNAs were significantly more abundant in UF-EVs, while 5 were less abundant compared to OF-EVs group, suggesting distinct roles in embryo-maternal communication. The most abundant miRNAs, included ocu-miR-148a-3p, ocu-let-7i-5p, and ocu-miR-10b-5p, related to embryo development and implantation. Target gene prediction analysis revealed that OF-EVs miRNAs are linked to MAPK, Hippo, TGF-β, Ras, PI3K-Akt, and mTOR pathways, while UF-EVs are associated with Hippo, PI3K-Akt, FoxO, endocytosis, and mTOR pathways, essential for cell proliferation, differentiation, immune regulation, and embryo-maternal interactions. Our findings suggest that EVs in rabbit reproductive fluids could play a fundamental role in early embryonic development. Future research should focus on the functional validation of these key candidate miRNAs, particularly those involved in reproductive physiology, offering promising insights for reproductive biotechnology and veterinary medicine.

This study aimed to investigate the effects of feeding broilers basal diets with high-temperature (35°C) stored soybean meal (SBM). A total of 192 one-day-old Arbor Acres Plus broilers (male) were divided into three groups with eight replicates for a 42-d experimental period. The three groups were fed a basal diet with fresh SBM, SBM stored at 35°C for 2 and 4 wks, respectively. The results showed that the protein carbonyl content of high-temperature stored SBM increased linearly (P = 0.083) and the protein free thiols content decreased (P = 0.021). Compared with the control group, SBM stored at high temperatures for 4 wks decreased the average daily gain (P = 0.002) and body weight (P = 0.002) of broilers, and increased the feed conversion ratio of 1-21 d (P = 0.008). The high-temperature stored SBM for 4 wks increased the 21-d ileal trypsin activity (P = 0.047), duodenal carbonyl content (P = 0.012), jejunal reactive oxygen species levels (P = 0.047) and 42-d duodenal and jejunal ROS levels of broilers (P = 0.029). The high-temperature stored SBM for 4 wks decreased the 21-d duodenal total superoxide dismutase activity (P = 0.003) and catalase activity (P = 0.006). Moreover, The high-temperature stored SBM altered intestinal chyme composition, increased 42-d jejunal methionine sulfoxide and cystine levels, and decreased L-leucine and lysine levels. These changes have caused 114 DEPs in the jejunum of broilers, there were 53 up-regulated and 61 down-regulated. Furthermore, the upregulated MAPK14 significantly affected the downstream transcription factors c-JUN and FoxO1. Our findings revealed the effects and mechanisms by which intestinal function was affected after broiler chickens ingested protein-oxidized SBM and identified the composition changes in intestinal chyme, DEPs and signaling pathways in the intestinal tissue.