WDR13 promotes the differentiation of bovine skeletal muscle‐derived satellite cells by affecting PI3K/AKT signaling

Abstract

Muscle satellite cells are usually at rest, and when externally stimulated or regulated, they can be further differentiated by cell fusion to form new myotubes and muscle fibers. WD repeat domain 13 (WDR13) is highly conserved in vertebrates. Studies have shown that mice lacking the Wdr13 gene develop mild obesity, hyperinsulinemia, and increased islet β cell proliferation. However, the role of WDR13 in bovine cells is unclear. Here, we investigated the effect of WDR13 on bovine skeletal muscle‐derived satellite cells (MDSCs). We found that WDR13 was upregulated in bovine MDSCs using western blotting and immunofluorescence experiments. Moreover, activation and inhibition of WDR13 expression increased and decreased cell differentiation, respectively, suggesting that WDR13 promotes bovine MDSC differentiation. To further understand the mechanism of action of WDR13, we examined changes in the PI3K/AKT signaling pathway following WDR13 activation or inhibition. In addition, cells were treated with a phosphoinositide kinase 3 (PI3K) inhibitor, LY294004, to observe cell differentiation. The results showed that activation of WDR13 inhibited the PI3K/AKT signaling pathway and enhanced cell differentiation. These data suggest that WDR13 can promote the differentiation of bovine MDSCs by affecting the PI3K/AKT signaling pathway.

SPARCL1 promotes C2C12 cell differentiation via BMP7-mediated BMP/TGF-β cell signaling pathway

“…Cells were co-transfected with 2 μg of the recombinant plasmid and 2 μg of pcDNA-dCas9-VP64 plasmid (AddGene, Middlesex, UK) for 72 h. Incubation of recombinant plasmid and SP-dCas9-VPR plasmid using Opti-MEM (Invitrogen Gibco, USA) as an incubation solution. The method used for cell transfection was described by Fu et al 25 when the cell density reached 70–80%, each well in a 6-well plate was added with 7.5 μl of Lipofectamine 2000 (Invitrogen Gibco, USA), 117.5 μl of opti-MEM, and 10 μl of SiRNA or NC (negative control), and the Lipofectamine 2000 with SiRNA or NC were co-transfected into the cells. In the following experiments, respective vectors are indicated as following: NC is the control group, VPR is the co-transfection vector of the activation group, pSPgRNA-S-2 is the vector for SPARCL1 activation, SiRNA-B is the SiRNA interference group for BMP7, and pSPgRNA-B-3 is an activation vector for BMP7.…”

WDR13: A Novel Gene Implicated in Non-Syndromic Intellectual Disability

“…Jeon, et al, showed that overexpression of FMRP alleviated cell death, increased Akt activity, and enhanced Bcl-xL production [ 25 ]. Studies in pancreatic cells have shown that the lack of the WDR13 protein activates the PI3K/Akt pathway [ 26 ]. With this in consideration, we hypothesize that a lack of WDR13 causes the over-activation of the PI3K/Akt cell signaling pathway, which results in increased expression of CAMK2A and FMRP.…”

Transcriptomic Profile of Primary Culture of Skeletal Muscle Cells Isolated from Semitendinosus Muscle of Beef and Dairy Bulls

“…It is likely that at least some of the tested cells isolated from semitendinosus muscle in LIM and HER breeds of cattle had already commenced differentiation which determines fusion into multinucleated myotubes, indicating increased expression of myogenin and other genes described in the following part of the Discussion, including myl2 , myh3 , actin alpha 1 skeletal muscle ( acta1 ), actin alpha cardiac muscle 1 ( actc1 ), tropomyosin 1 ( tpm1 ), troponin T2, cardiac type ( tnnt2 ), and troponin C type 1, slow ( tnnc1 ). These genes play a key role in the differentiation and fusion of muscle cells, and also have a positive impact on the process of myogenesis of skeletal muscles [ 35 , 36 , 37 , 38 ].…”

“…On the contrary, lowered expression of MYH3 and MYL2 was shown in muscle cells deprived of myogenin [ 33 ]. Myosin heavy chains 3 (MYH3) undergoes expression in skeletal muscle cells, promotes cell fusion into myotubes [ 35 , 55 ], and its expression is positively correlated with higher growth of muscle mass in cattle [ 56 , 57 ]. Similar relationships were observed in the present study, with a higher myh3 expression level in beef bulls ( Figure 4 , Table 1 ).…”