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Effects of α-enolase gene silencing on reproductive-related hormone receptor expression and steroid hormone synthesis of primary granulosa cells from goose F1 follicles Cover

Effects of α-enolase gene silencing on reproductive-related hormone receptor expression and steroid hormone synthesis of primary granulosa cells from goose F1 follicles

Journal of Veterinary Research
Volume 64 (2020): Issue 1 (March 2020)
By: Hong Ji,  Chun-Yang Niu,  Hong-Liang Zhang,  Jing-Ru Guo,  Li Zhen,  Shuai Lian,  Chuang Yang,  Huan-Min Yang and  Jian-Fa Wang  
Open Access
|Jan 2020

Figures & Tables

Fig. 1

Transfection of shRNA-ENO1 in granulosa cells and detection of ENO1 expression. A – transfection of shRNA-ENO1 in granulosa cells (100 ×) including granulosa cells under ordinary light and fluorescence. B – ENO1 mRNA expression by qRT-PCR in granulosa cells transfected with shRNA-ENO1 after 48 h. ENO1 mRNA expression by qRT-PCR in granulosa cells was measured by qRT-PCR. C – ENO1 expression by Western blot in granulosa cells transfected with shRNA-ENO1 after 48 h. The significance of difference for the ENO1 expression level among the different groups was determined by Student’s t-test. ** P < 0.01, and * P < 0.05 when compared with the control sample
Transfection of shRNA-ENO1 in granulosa cells and detection of ENO1 expression. A – transfection of shRNA-ENO1 in granulosa cells (100 ×) including granulosa cells under ordinary light and fluorescence. B – ENO1 mRNA expression by qRT-PCR in granulosa cells transfected with shRNA-ENO1 after 48 h. ENO1 mRNA expression by qRT-PCR in granulosa cells was measured by qRT-PCR. C – ENO1 expression by Western blot in granulosa cells transfected with shRNA-ENO1 after 48 h. The significance of difference for the ENO1 expression level among the different groups was determined by Student’s t-test. ** P < 0.01, and * P < 0.05 when compared with the control sample

Fig. 2

Quantitative RT-PCR–detected expression of FSHR, LHR, ERα, ERβ, GHR, and IGFBP-1 mRNA in the granulosa transfected with shRNA-ENO1 after 48 h detected by qRT-PCR. A – FSHR, B – LHR, C – GHR, D – IGFBP-1, E – ERα, F – ERβ. The significance of difference for the hormone receptor levels were determined by Student’s t-test. ** p < 0.01, and * p < 0.05 when compared with the control sample
Quantitative RT-PCR–detected expression of FSHR, LHR, ERα, ERβ, GHR, and IGFBP-1 mRNA in the granulosa transfected with shRNA-ENO1 after 48 h detected by qRT-PCR. A – FSHR, B – LHR, C – GHR, D – IGFBP-1, E – ERα, F – ERβ. The significance of difference for the hormone receptor levels were determined by Student’s t-test. ** p < 0.01, and * p < 0.05 when compared with the control sample

Fig. 3

ELISA-detected concentration of six hormones in the supernatant of granulosa transfected with shRNA-ENO1 after 48 h. A – oestradiol. B – activin. C – progesterone. D – testosterone. E – inhibin. F – follistatin. The significance of difference for the hormone levels was determined by Student’s t-test. ** P < 0.01, and * P < 0.05 when compared with the control sample
ELISA-detected concentration of six hormones in the supernatant of granulosa transfected with shRNA-ENO1 after 48 h. A – oestradiol. B – activin. C – progesterone. D – testosterone. E – inhibin. F – follistatin. The significance of difference for the hormone levels was determined by Student’s t-test. ** P < 0.01, and * P < 0.05 when compared with the control sample

List of primers used for quantitative RT-PCR

Target genePrimer sequence (5′–3′)Amplicon size (bp)
ENO1F: GCGGTGCCTCAACTGGAATT 183
R: CCATGTCCAGCATCAGTTTGTC
GAPDH

F: GCTGATGCTCCCATGTTCGTGAT

R: GTGGTGCAAGAGGCATTGCTGAC		86
LHR

F: GTAACACTGGAATAAGGGAAT

R: GAAGGCTTGACTGTGGATA		191
ERα

F: ACCCAAACAGACCATTCAACGAA

R: CGCCAGACTAAGCCAATCATCAG		187
ERβ

F: AAGTGGGAATGATGAAATGTGGC

R: GGACTGACCGTGCTGAGGAGAAT		163
FSHR

F: TCCTGTGCTAACCCTTTCCTCTA

R: AACCAGTGAATAAATAGTCCCATC		207
IGFBP-1

F: CCTTGTCAGAAGGAGCTCTA

R: CATCCAGTGAAGTTTCACACT		145
GHR

F: GCCCCTGCTGACATTTGAGAAT

R: GGCCACTGCAGAAGATCATCAC		115
DOI: https://doi.org/10.2478/jvetres-2020-0008 | Journal eISSN: 2450-8608
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Language: English
Page range: 141 - 149
Submitted on: Jun 6, 2019
Accepted on: Jan 17, 2020
Published on: Jan 31, 2020
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
Zi goose,
follicular granulosa cell,
α-enolase,
hormone receptor,
hormone
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2020 Hong Ji, Chun-Yang Niu, Hong-Liang Zhang, Jing-Ru Guo, Li Zhen, Shuai Lian, Chuang Yang, Huan-Min Yang, Jian-Fa Wang, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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