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Chaperone-mediated autophagy plays an important role in regulating retinal progenitor cell homeostasis.
- Source :
- Stem Cell Research & Therapy; 4/1/2022, Vol. 13 Issue 1, p1-14, 14p
- Publication Year :
- 2022
-
Abstract
- Purpose: To explore the function and regulatory mechanism of IFITM3 in mouse neural retinal progenitor cells (mNRPCs), which was found to be very important not only in the development of the retina in embryos but also in NRPCs after birth. Methods: Published single-cell sequencing data were used to analyze IFITM3 expression in mNRPCs. RNA interference was used to knock down the expression of IFITM3. CCK-8 assays were used to analyze cell viability. RNA-seq was used to assess mRNA expression, as confirmed by real-time quantitative PCR, and immunofluorescence assays and western blots were used to validate the levels of relative proteins, and autophagy flux assay. Lysosomal trackers were used to track the organelle changes. Results: The results of single-cell sequencing data showed that IFITM3 is highly expressed in the embryo, and after birth, RNA-seq showed high IFITM3 expression in mNRPCs. Proliferation and cell viability were greatly reduced after IFITM3 was knocked down. The cell membrane system and lysosomes were dramatically changed, and lysosomes were activated and evidently agglomerated in RAMP-treated cells. The expression of LAMP1 was significantly increased with lysosome agglomeration after treatment with rapamycin (RAMP). Further detection showed that SQSTM1/P62, HSC70 and LAMP-2A were upregulated, while no significant difference in LC3A/B expression was observed; no autophagic flux was generated. Conclusion: IFITM3 regulates mNRPC viability and proliferation mainly through chaperone-mediated autophagy (CMA) but not macroautophagy (MA). IFITM3 plays a significant role in maintaining the homeostasis of progenitor cell self-renewal by sustaining low-level activation of CMA to eliminate deleterious factors in cells. [ABSTRACT FROM AUTHOR]
- Subjects :
- PROGENITOR cells
AUTOPHAGY
CELL survival
GENE expression
LYSOSOMES
Subjects
Details
- Language :
- English
- ISSN :
- 17576512
- Volume :
- 13
- Issue :
- 1
- Database :
- Complementary Index
- Journal :
- Stem Cell Research & Therapy
- Publication Type :
- Academic Journal
- Accession number :
- 156108388
- Full Text :
- https://doi.org/10.1186/s13287-022-02809-z