Background Muscle development is a precisely orchestrated and complex process, and circular RNAs (circRNAs) has been demonstrated to play important roles in skeletal muscle growth and development. However, the regulatory functions of circRNA during buffalo muscle developmental processes have not been understood.
Results In this study, Ribo-Zero RNA-Seq was performed to investigate the circRNAs expression profiles of proliferated and differentiated buffalo myoblasts. A stringent set of 3,142 circRNAs was finally characterized. Comparing the expression profiles of circRNAs revealed that 110 circRNAs were expressed differentially during myoblast differentiation. We focused on the role of a candidate circRNA, which was named circPICALM based on its host gene PICALM, and was highly (but differentially) expressed in proliferated and differentiated myoblasts. Flow cytometry, EdU incorporation, and Western blotting assays demonstrate that circPICALM promoted myoblasts proliferation and inhibited cells apoptosis. Moreover, overexpression of circPICALM promoted the differentiation of primary buffalo myoblasts. Moreover, circPICALM in vivo stimulated skeletal muscle regeneration in cardiotoxin-induced muscle injury. The RNA pulldown results showed that circPICALM could capture TUBA1B protein, revealing that circPICALM might exert its biological function by binding TUBA1B protein.
Conclusions These results demonstrate that the novel non-coding regulator circPICALM induces myoblast differentiation and skeletal muscle regeneration.

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9
This is a list of supplementary files associated with this preprint. Click to download.
Loading...
Posted 11 May, 2020
Posted 11 May, 2020
Background Muscle development is a precisely orchestrated and complex process, and circular RNAs (circRNAs) has been demonstrated to play important roles in skeletal muscle growth and development. However, the regulatory functions of circRNA during buffalo muscle developmental processes have not been understood.
Results In this study, Ribo-Zero RNA-Seq was performed to investigate the circRNAs expression profiles of proliferated and differentiated buffalo myoblasts. A stringent set of 3,142 circRNAs was finally characterized. Comparing the expression profiles of circRNAs revealed that 110 circRNAs were expressed differentially during myoblast differentiation. We focused on the role of a candidate circRNA, which was named circPICALM based on its host gene PICALM, and was highly (but differentially) expressed in proliferated and differentiated myoblasts. Flow cytometry, EdU incorporation, and Western blotting assays demonstrate that circPICALM promoted myoblasts proliferation and inhibited cells apoptosis. Moreover, overexpression of circPICALM promoted the differentiation of primary buffalo myoblasts. Moreover, circPICALM in vivo stimulated skeletal muscle regeneration in cardiotoxin-induced muscle injury. The RNA pulldown results showed that circPICALM could capture TUBA1B protein, revealing that circPICALM might exert its biological function by binding TUBA1B protein.
Conclusions These results demonstrate that the novel non-coding regulator circPICALM induces myoblast differentiation and skeletal muscle regeneration.

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9
This is a list of supplementary files associated with this preprint. Click to download.
Loading...