Effects of N-carbamylglutamate and L-arginine on steroidogenesis and gene expression in bovine granulosa cells

Feng, Tao; Schütz, Luís F.; Morrell, B.C.; Perego, Maria Chiara; Spicer, L. J.

doi:10.1016/j.anireprosci.2017.11.012

Cited by 17 publications

(10 citation statements)

References 43 publications

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“…Ovaries from non-pregnant beef cows were collected from a local abattoir and based on surface diameter, GC was collected and prepared from small (1 to 5 mm) and large (8 to 22 mm) follicles as previously described [ 24 , 44 ]. The viability of GC from small and large follicles was determined by the trypan blue exclusion method and averaged 58 ± 7% and 64 ± 6%, respectively, and is similar to viabilities reported in previous studies [ 45 , 46 , 47 , 48 ].…”

Section: Methodssupporting

confidence: 82%

In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells

Chiminelli

Spicer

Maylem

et al. 2022

Toxins

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The emerging Fusarium mycotoxins enniatins (ENNs) have been the focus of new research because of their well-documented existence in various cereal and grain products. Research findings indicate that reproductive disorders may be caused by exposure to Fusarium mycotoxins, but little work has evaluated ENNs on reproductive function. Therefore, to determine the effects of ENNA on the proliferation and steroidogenesis of granulosa cells (GC), experiments were conducted using bovine GC cultures. In vitro, ENNA (1–5 μM) inhibited (p < 0.05) hormone-induced GC progesterone and estradiol production. The inhibitory effect of ENNA on estradiol production was more pronounced in small- than large-follicle GC. In large-follicle GC, 0.3 μM ENNA had no effect (p > 0.10) whereas 1 and 3 μM ENNA inhibited GC proliferation. In small-follicle GC, ENNA (1–5 μM) dramatically decreased (p < 0.05) GC proliferation. Using cell number data, the IC50 of ENNA was estimated at 2 μM for both follicle sizes. We conclude that ENNA can directly inhibit ovarian function in cattle, decreasing the proliferation and steroid production of GC.

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Section: Methodssupporting

confidence: 82%

In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells

Chiminelli

Spicer

Maylem

et al. 2022

Toxins

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“…More recently, several results have been reported that gilts with dietary supplementation of 0.05% and 0.1% of NCG had more pigs born alive, and significantly increased the live litter weight [10,11]. A similar effect of NCG on growth promotion and reproductive performance enhancement have also been found in other species of animals including the rat [12], cattle [13], sheep [14], goat [15], and yellow-feather broiler [16].…”

Section: Introductionmentioning

confidence: 75%

Determination of N-Carbamylglutamate in Feeds and Animal Products by High Performance Liquid Chromatography Tandem Mass Spectrometry

Zeng

Kong

et al. 2019

Molecules

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N-carbamylglutamate (NCG), a synthetic analogue of N-acetylglutamate, is an activator of blood ammonia conversion and endogenous arginine synthesis. Here, we established an accurate quantitative determination of NCG in feeds, animal tissues, and body fluids using the high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The sample pretreatment procedures included extraction with 0.5% of formic acid in water/methanol (80/20, v/v), and purification using an anionic solid phase extraction cartridge. Satisfactory separation of NCG was achieved in 20 min with the application of an Atlantis T3 column, and a confirmative detection of NCG was ensured by multiple reaction monitoring of positive ions. NCG spiked in feeds, tissues, and body fluids were evaluated in regard to linearity, sensitivity, recovery, and repeatability. Recoveries for different sample matrices were in the range of 88.12% to 110.21% with relative standard deviations (RSDs) less than 8.8%. Limits of quantification were within the range of 0.012 to 0.073 mg kg−1 and 0.047 to 0.077 μg mL−1 for solid and liquid samples, respectively. This study will provide a solid foundation for the evaluation of availability and metabolic mechanism of NCG in animals.

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“…The total RNA extracted was reverse-transcribed into cDNA in a 10 μL reaction system by using a Script cDNA Synthesis Kit (Bio-Rad), and then the first-strand cDNA was directly used for RT-qPCR or stored at −80 °C [ 34 ].…”

Section: Methodsmentioning

confidence: 99%

“…The PCR procedure was the same as previously described [ 32 ]. Additionally, two negative controls, namely a no-template control and a no-reverse-transcriptase control, were included to confirm the absence of contaminants in the master mix and DNA contamination in RNA, respectively [ 34 ]. The relative abundance of the target mRNA transcripts was calculated as 2 − ∆∆Ct using the comparative threshold cycling method, normalized to GAPDH ribosomal RNA levels as previously described [ 34 , 35 ].…”

Section: Methodsmentioning

confidence: 99%

Progesterone Induces Apoptosis and Steroidogenesis in Porcine Placental Trophoblasts

Liu

Ding

Yang

et al. 2022

Animals

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Placentation and placental steroidogenesis are important for pregnancy and maternal–fetal health. As pregnancy progresses, the main site of progesterone (P4) synthesis changes from the corpus luteum to the placenta, in which placental trophoblasts are the main cell type for P4 synthesis. Therefore, this study investigated the effects of P4 on apoptosis and steroidogenesis in porcine placental trophoblasts and the underlying molecular mechanisms. Porcine placental trophoblasts were treated with different concentrations of P4 for 48 h in a serum-free medium in vitro. Cell number, steroidogenesis, and relevant gene and protein expression levels were detected. A high dose of P4 (10.0 μM) significantly increased P4 (p < 0.01), androstenedione (p < 0.05), testosterone (p < 0.05), and estradiol (p < 0.05) production in porcine placental trophoblasts compared with that in control cells, while a low dose of P4 (1 × 10−3 μΜ) had no marked impact on steroid production. The mRNA expression of apoptosis-related genes (CASP3, CASP8, and Bax) (p < 0.05) and steroidogenesis-related genes (CYP11A1, CYP19A1, and StAR) (p < 0.01) was upregulated, and the expression of HSD3B and HSD17B4 was inhibited (p < 0.05) in the porcine placental trophoblasts treated with high doses of P4. Low doses of P4 had a lighter effect on gene expression than high doses. The expression of apoptosis-related proteins CASP3 (p < 0.05), and Bax (p < 0.01) and steroidogenesis-related proteins CYP19A1 (p < 0.05) and StAR (p < 0.01) was raised, but the proliferation-related protein CCND2 (p < 0.01) was downregulated in the pTr cells treated with high dose of P4. In comparison, a low dose of P4 inhibited the expression of Bax, CYP11A1 (all p < 0.01), and CCND2 (p < 0.05), but the expression of CASP3 (p < 0.05) and StAR (p < 0.01) was upregulated. In summary, excessive P4 can induce the apoptosis of porcine placental trophoblasts and lead to abnormal steroidogenesis in the placenta and hormone imbalance.

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Effects of N-carbamylglutamate and L-arginine on steroidogenesis and gene expression in bovine granulosa cells

Cited by 17 publications

References 43 publications

In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells

In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells

Determination of N-Carbamylglutamate in Feeds and Animal Products by High Performance Liquid Chromatography Tandem Mass Spectrometry

Progesterone Induces Apoptosis and Steroidogenesis in Porcine Placental Trophoblasts

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