miR-21在鸭胚胎期胸肌发育中的功能研究

doi:10.11843/j.issn.0366-6964.2019.02.007

Abstract

Abstract:

The molecular characteristics, the spatiotemporal expression patterns and the function of miR-21 during the proliferation and differentiation of duck skeletal muscle satellite cells were studied to provide support for exploring the mechanism of skeletal muscle development. One hundred and eighty eggs with the weight of (98 ±5) g were hatched under the same conditions. The embryo breast muscle samples were collected from 3 hatched ducks from 11^th day to 27^th day of hatching (E11, E12,E13……E26,E27) under sterile condition, respectively. The leg muscle, heart, liver, kidney, stomach, small intestine, abdominal fat and sebum were sampled from hatched eggs on E27 under sterile condition. The skeletal muscle satellite cells were isolated and purified by mixed enzyme digestion and differential adherence methods. The role of miR-21 was explored by fluorescence quantitative analysis, protein hybridization, EdU detection and flow cytometry during the proliferation and differentiation of duck skeletal muscle satellite cells transfected with Agomir and Antagomir of miR-21. The results of qRT-PCR showed that the expression of miR-21 increased gradually with the increase of incubation age, reached the highest level on E19, and then decreased gradually. In addition, the highest level of miR-21 expression was found in liver on E27. miR-21 was overexpressed in the proliferating duck skeletal muscle satellite cells, the mRNA expression levels of CDK2 and CyclinD1 significantly (P<0.05) and extremely significantly increased (P<0.01), respectively. The number of positive cells detected by EdU was significantly increased (P<0.05). The number of G2 phase cells was extremely significantly decreased (P<0.01), and the number of S phase cells was significantly increased (P<0.05) by flow cytometry analysis. When the expression of miR-21 was suppressed in the proliferating duck skeletal muscle satellite cells, the results were contrary to that of miR-21 overexpression in the proliferating duck skeletal muscle satellite cells. When the miR-21 was overexpressed in the differentiated duck skeletal muscle satellite cells, the mRNA expression of MyoG and MyHC were extremely significantly increased (P<0.01) by qRT-PCR. The protein expression of MyoG was siginificantly increased(P<0.05), and the protein expression of MyHC was extremely significantly increased (P<0.01) by Western blot (WB) analysis. Both the number of MyHC positive cells and the number of myotubes containing more than 10 nuclei were extremely significantly increased (P<0.01) detected by immunofluorescence assay. The results were contrary to that of miR-21 was suppressed in the differentiated duck skeletal muscle satellite cells. The results indicate that miR-21 can promote the proliferation and differentiation of skeletal muscle satellite cells of myoblasts of ducks.

CLC Number:

S834.2

GU Lihong, XU Tieshan, LIN Zhemin, ZHAO Jianguo, LIN Peng, YANG Shaoxiong, LIN Dajie, HOU Shuisheng. Function of miR-21 in Embryonic Breast Muscle Development of Duck[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 287-301.

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Function of miR-21 in Embryonic Breast Muscle Development of Duck

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