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Regulation of Glucose Phosphate Isomerase by the 3'UTR-Specific miRNAs miR-302b and miR17-5p in Chicken Primordial Germ Cells

Cited 19 time in Web of Science Cited 18 time in Scopus
Authors
Rengaraj, Deivendran; Park, Tae Sub; Lee, Sang In; Lee, Bo Ram; Han, Beom Ku; Song, Gwonhwa; Han, Jae Yong
Issue Date
2013
Publisher
Society for the Study of Reproduction
Citation
Biology of Reproduction, vol.89 no.2, pp. 1-11
Keywords
chickengene regulationglucose phosphate isomerase (GPI)metabolismmicroRNAprimordial germ cells (PGCs)target efficiency
Abstract
Glucose phosphate isomerase (GPI) involves in the reversible isomerization of glucose-6-phosphate to fructose-6-phosphate in glucose pathways. Because glucose metabolism is crucial for the proliferation and differentiation of embryonic stem and germ cells, reducing GPI expression may affect the characteristic features of these cells. MicroRNAs (miRNAs) have been shown to regulate genes. In the present study, we investigated the regulation of chicken GPI by its predicted miRNAs. We determined the expression patterns of seven GPI 3′-untranslated region (3′UTR)-targeting miRNAs, including the gga-miR-302 cluster, gga-miR-106, gga-miR-17-5p, and gga-miR-20 cluster in chicken primordial germ cells (PGCs), compared with GPI mRNA. Among the miRNAs, gga-miR-302b, gga-miR-302d, and gga-miR-17-5p were expressed at lower levels than GPI mRNA. The remaining four miRNAs—gga-miR-302c, gga-miR-106, gga-miR-20a, and gga-miR-20b—were expressed at higher levels than the expression of GPI mRNA. Next, we cotransfected four candidate miRNAs—gga-miR-302b, gga-miR-106, gga-miR-17-5p, and gga-miR-20a—with GPI 3′UTR into 293FT cells by dual fluorescence reporter assay. Overexpression of gga-miR-302b and gga-miR-17-5p miRNAs in 293FT cells significantly downregulated GPI expression, whereas the other two miRNAs had no effect. Then, knockdown and overexpression of these four candidate miRNAs were performed by RNA interference assay to regulate GPI in PGCs. In the RNA interference assay, the expression of GPI was greatly regulated by gga-miR-302b and gga-miR-17-5p. Finally, we examined the effects of GPI regulation on PGC proliferation and migration. Our results suggested that the regulation of GPI by gga-miR-302b and gga-miR-17-5p affected PGCs proliferation. However, regulation of GPI using these two miRNAs did not affect the migration of PGCs into embryonic gonads.
ISSN
0006-3363
Language
English
URI
https://hdl.handle.net/10371/100367
DOI
https://doi.org/10.1095/biolreprod.112.105692
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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