Cathepsin D knockdown regulates biological behaviors of granulosa cells and affects litter size traits in goats

Zhou, Zhinan; Chen, Xiang; Zhu, Min; Wang, Weiwei; Ao, Zheng; Zhao, Jianjian; Tang, Wen; Lei, Hong

doi:10.1631/jzus.b2100366

Cited by 9 publications

(12 citation statements)

References 44 publications

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“…The right ovary was fixed in 4% paraformaldehyde solution and was used for immunohistochemistry and HE staining. The granulosa cells were also identified through cellular immunofluorescence for FSHR-specific expression of protein staining [ 24 ]. All animal experiments were approved by the Animal Ethics Committee of Guizhou University (Guiyang, China).…”

Section: Methodsmentioning

confidence: 99%

“…Immunohistochemistry was performed according to a previously described protocol [ 24 ]. Ovary tissue sections (5 mm) were first fixed and embedded in paraffin.…”

Section: Methodsmentioning

confidence: 99%

“…All data are presented as means ± SD of three biological replicates and three technical replicates to ensure the accuracy of the experimental data. The mRNA levels of target genes were normalized with a housekeeping gene ( β-actin , 139 bp) and expressed as 2 −∆∆CT using samples on pEGFP-N3 or sh-NC as a calibrator [ 24 ]. The significance threshold was set at p < 0.05.…”

Section: Methodsmentioning

confidence: 99%

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CYP19A1 May Influence Lambing Traits in Goats by Regulating the Biological Function of Granulosa Cells

Zhang

Chen

Zhou

et al. 2022

Animals

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Abnormal expression of CYP19A1, a gene related to steroid hormone synthesis, causes steroid hormone disruption and leads to abnormal ovulation in granulosa cells. However, the exact mechanism of CYP19A1 regulation is unclear. In this study, we confirmed the localization of CYP19A1 in goat ovarian tissues using immunohistochemistry. Subsequently, we investigated the effects of CYP19A1 on granulosa cell proliferation, steroid hormone secretion, and expression of candidate genes for multiparous traits by overexpressing and silencing CYP19A1 in goat granulosa cells (GCs). The immunohistochemistry results showed that CYP19A1 was expressed in all types of follicular, luteal, and granulosa cells, with subcellular localization results revealing that CYP19A1 protein was mainly localized in the cytoplasm and nucleus. Overexpression of CYP19A1 significantly increased the mRNA levels of CYP19A1, FSHR, and INHBA, which are candidate genes for multiple birth traits in goats. It also promoted cell proliferation, PCNA and Cyclin E mRNA levels in granulosa cells, and secretion of estrogen and progesterone. However, it inhibited the mRNA levels of STAR, CYP11A1, and 3βSHD, which are genes related to steroid synthesis. Silencing CYP19A1 expression significantly reduced CYP19A1, FSHR, and INHBA mRNA levels in granulosa cells and inhibited granulosa cell proliferation and PCNA and Cyclin E mRNA levels. It also reduced estrogen and progesterone secretion but enhanced the mRNA levels of STAR, CYP11A1, and 3βSHD. CYP19A1 potentially influenced the lambing traits in goats by affecting granulosa cell proliferation, hormone secretion, and expression of candidate genes associated with traits for multiple births.

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Section: Methodsmentioning

confidence: 99%

“…Immunohistochemistry was performed according to a previously described protocol [ 24 ]. Ovary tissue sections (5 mm) were first fixed and embedded in paraffin.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

CYP19A1 May Influence Lambing Traits in Goats by Regulating the Biological Function of Granulosa Cells

Zhang

Chen

Zhou

et al. 2022

Animals

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“…Immunohistochemistry was performed as described previously [ 33 , 34 ]. In short, paraffin sections of 5-μm thickness were attached to microscope slides, heated at 65 °C for 2 h, dewaxed in xylene, and then rehydrated in a graded series of ethanol.…”

Section: Methodsmentioning

confidence: 99%

“…ImageJ (18.0) software was used to calculate the grey value. SPSS (26.0) software was used for one-way ANOVA to calculate the differential expression values [ 34 ].…”

Section: Methodsmentioning

confidence: 99%

A Proteomic Study of the Effect of N-acetylcysteine on the Regulation of Early Pregnancy in Goats

Yang

Chen

Tian

et al. 2022

Animals

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Dietary supplementation with N-acetyl-L-cysteine (NAC) may support early pregnancy regulation and fertility in female animals. The purpose of this study was to investigate the effect of supplementation with 0.07% NAC on the expression of the uterine keratin gene and protein in Qianbei-pockmarked goats during early pregnancy using tandem mass spectrometry (TMT) relative quantitative proteomics. The results showed that there were significant differences in uterine keratin expression between the experimental group (NAC group) and the control group on day 35 of gestation. A total of 6271 proteins were identified, 6258 of which were quantified by mass spectrometry. There were 125 differentially expressed proteins (DEPs), including 47 upregulated and 78 downregulated proteins, in the NAC group. Bioinformatic analysis showed that these DEPs were mainly involved in the transport and biosynthesis of organic matter and were related to the binding of transition metal ions, DNA and proteins and the catalytic activity of enzymes. They were enriched in the Jak-STAT signalling pathway, RNA monitoring pathway, amino acid biosynthesis, steroid biosynthesis and other pathways that may affect the early pregnancy status of does through different pathways and thus influence early embryonic development. Immunohistochemistry, real-time quantitative PCR and Western blotting were used to verify the expression and localization of glial fibrillary acidic protein (GFAP) and pelota mRNA surveillance and ribosomal rescue factor (PELO) in uterine horn tissue. The results showed that both PELO and GFAP were localized to endometrial and stromal cells, consistent with the mass spectrometry data at the transcriptional and translational levels. Moreover, NAC supplementation increased the levels of the reproductive hormones follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol (E2), progesterone (P4), superoxide dismutase (SOD), glutamate peroxidase (GSH-Px) and nitric oxide (NO) in the serum of does. These findings provide new insight into the mechanism by which NAC regulates early pregnancy and embryonic development in goats.

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Global gene expression analysis reveals a subtle effect of DEHP in human granulosa cell line HGrC1

Nenadov

Tesic

Tomanic

et al. 2023

Reproductive Toxicology

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Cathepsin D knockdown regulates biological behaviors of granulosa cells and affects litter size traits in goats

Cited by 9 publications

References 44 publications

CYP19A1 May Influence Lambing Traits in Goats by Regulating the Biological Function of Granulosa Cells

CYP19A1 May Influence Lambing Traits in Goats by Regulating the Biological Function of Granulosa Cells

A Proteomic Study of the Effect of N-acetylcysteine on the Regulation of Early Pregnancy in Goats

Global gene expression analysis reveals a subtle effect of DEHP in human granulosa cell line HGrC1

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