This study was conducted to clarify the effect of the inhibiting action of inhibin on porcine granulosa cell proliferation and function, and to investigate the underlying intracellular regulatory molecular mechanisms. Porcine granulosa cells were cultured in vitro, and were treated with an anti-inhibin alpha subunit antibody, with or without co-treatment of follicle-stimulating hormone (FSH) in the culture medium. Treatment with anti-inhibin alpha subunit antibody led to a significant increase in estradiol (E2) secretion and cell proliferation. Anti-inhibin alpha subunit antibody worked synergistically with FSH at low concentrations (25 microg/mL) to stimulate E2 secretion, but attenuated FSH action at high concentrations (50 microg/mL). Immunoneutralization of inhibin bioactivity increased FOXL2, Smad3, and PKA phosphorylation, and mRNA expression of the transcription factors CEBP and c-FOS. The expression of genes encoding gonadotropin receptors, FSHR and LHR, and of those involved in steroidogenesis, as well as IGFs and IGFBPs, the cell cycle progression factors cyclinD1 and cyclinD2, and the anti-apoptosis and anti-atresia factors Bcl2, TIMP, and ADAMTS were upregulated following anti-inhibin alpha-subunit treatment. Treatment with anti-inhibin alpha subunit down regulated expression of the pro-apoptotic gene encoding caspase3. Although expression of the pro-angiogenesis genes FN1, FGF2, and VEGF was upregulated, expression of the angiogenesis-inhibiting factor THBS1 was downregulated following anti-inhibin alpha subunit treatment. These results suggest that immunoneutralization of inhibin bioactivity, through augmentation of the activin and gonadotropin receptor signaling pathways and regulation of gene expression, permits the development of healthy and viable granulosa cells. These molecular mechanisms help to explain the enhanced ovarian follicular development observed following inhibin immunization in animal models.

The poultry ovary is a preferred target for E. coli and Salmonella infection of tissues, and lipopolysaccharide (LPS) is a critical molecule in infecting the organism and interfering with cell function, invading the ovaries through the cloaca and interfering with progesterone (P4) secretion by follicular granulosa cells (GCs), seriously affecting the health of breeding geese. miRNAs are small, non-coding RNAs with a variety of important regulatory roles. To investigate the mechanism of miR-10a-5p mediated LPS inhibition of progesterone synthesis in goose granulosa cells, Yangzhou geese at peak laying period were selected as experimental animals to verify the expression levels of genes and transcription factors related to progesterone synthesis. In this study, bioinformatic predictions identified miR-10a-5p target gene CYP11A1, and genes and transcription factors related to the sex steroid hormone secretion pathway were screened. We detected that LPS inhibited CYP11A1 expression while increasing miR-10a-5p expression in vivo. Progesterone decreased significantly in goose granulosa cells treatment with 1 μg/mL LPS for 24 h, while progesterone-related genes and regulatory factors were also suppressed. We also determined that the downregulation of miR-10a-5p led to CYP11A1 expression. Overexpression of miR-10a-5p suppressed LPS-induced CYP11A1 expression, resulting in decreased progesterone secretion. Our findings indicated that miR-10a-5p was up-regulated by LPS and inhibited progesterone synthesis by down-regulating CYP11A1. This study provides insight into the molecular mechanisms regulating geese reproduction and ovulation.

Chronic heat stress (CHS) has been exerting great pressure on the poultry industry due to the global warming. CHS-induced negative impacts deeply change the physiology, metabolism, egg production and meat quality of ducks. To reduce the increasing economic loss, different environmental strategies have been adopted. Here, we comprehensively compared three rearing modes for Cherry-Valley ducks (CVds) mainly by changing the environmental temperature. Plus, the transcriptome analysis and physiological anatomy were implied to investigate the underlying molecular biology and pneumonocyte dynamics. It turns out that the CVds affected by CHS have poor growth performance and bad carcass traits. In addition, the irreversible damage caused by CHS in the lung tissue of CVds was observed. Furthermore, several potential genes related to CHS have been found, such as HSPA8, IGF1, FGFR1, ACSL3, ACSL6, NR1H3 and GAPDH. This study strongly indicates the severe injury of CHS from the high-temperature environment and points out one of the adopting directions for duck rearing.