Knockdown of Death-Associated Protein Expression Induces Global Transcriptome Changes in Proliferating and Differentiating Muscle Satellite Cells
| Autor: | Kelly R. B. Sporer, Robert J. Tempelman, Sandra G. Velleman, Yuwares Malila, Katherine A. Horton, Gale M. Strasburg, Kent M. Reed |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Physiology muscle lcsh:Physiology Transcriptome 03 medical and health sciences 0302 clinical medicine Physiology (medical) Gene expression medicine turkey Mechanistic target of rapamycin PI3K/AKT/mTOR pathway Original Research satellite cells Gene knockdown biology lcsh:QP1-981 Microarray analysis techniques Skeletal muscle death-associated protein Protein ubiquitination Cell biology 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis global gene expression biology.protein muscle development microarray |
| Zdroj: | Frontiers in Physiology, Vol 11 (2020) Frontiers in Physiology |
| Popis: | Death-associated protein (DAP) undergoes substantial changes in expression during turkey skeletal muscle development, decreasing from the 18 day embryonic stage to 1 day posthatch, and again from 1 day posthatch to 16 weeks of age. These changes suggest that DAP plays an important role at critical stages of the developmental process. The objective of this study was to elucidate the role of DAP in muscle development by examining the effect of reduced DAP expression on global gene expression in proliferating and differentiating turkey pectoralis major muscle satellite cells. Small interfering RNA was used to knock down expression of DAP and the transcriptome was subsequently profiled using a turkey skeletal muscle long oligonucleotide microarray. Microarray data were corroborated using quantitative real-time PCR. In proliferating cells, 458 loci, resulting in 378 uniquely annotated genes, showed differential expression (false discovery rate, FDR < 0.05). Pathway analysis highlighted altered eukaryotic translational initiation factors (eIFs) signaling, protein ubiquitination, sirtuin signaling, and mechanistic target of rapamycin (mTOR) signaling as the primary pathways affected in the knockdown proliferating cells. The findings underpinned the potential DAP involvement in cell proliferation of turkey satellite cells through the coordination between protein synthesis and cell cycle. In differentiating cells, 270 loci, accounting for 189 unique genes, showed differential expression (FDR < 0.05). Decreased expression of genes encoding various myofibrillar proteins and proteins involved in sarcoplasmic reticulum calcium flux suggests that DAP may affect regulation of calcium homeostasis and cytoskeleton signaling. This study provides the first evidence that reduced expression of DAP significantly alters the transcriptome profile of pectoralis major muscle satellite cells, thereby reducing proliferation and differentiation. |
| Databáze: | OpenAIRE |
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