Characterization of POU5F1 (OCT4) gene and its promoter in buffalo ESC-like cells identifies multiple transcription start sites and expression of four pseudogenes

Singh, Narendra Kumar; George, Aman; Sharma, Ruchi; Singla, S. K.; Palta, P.; Manik, R. S.; Chauhan, M. S.; Singh, Dheer

doi:10.1016/j.gene.2011.09.036

Cited by 5 publications

(2 citation statements)

References 30 publications

(37 reference statements)

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“…A similar finding for NANOG protein was reported in pig testis, where a truncated protein of smaller size was observed [30]. In a recent report, Western blot of POU5F1 revealed presence of four distinct visible bands in buffalo ESC-like cells, with the upper most band (∼38 kD) corresponding to the parent POU5F1 and the other three proteins (size ranging from 35.5 kDa to <24 kDa) resulting from pseudogene expression, leading to the production of truncated proteins [47]. In the present study, the uppermost band of POU5F1 protein was of much larger size (i.e., ∼47 kDA).…”

Section: Discussionsupporting

confidence: 83%

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Spermatogonial Stem Cells (SSCs) in Buffalo (Bubalus bubalis) Testis

2012

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BackgroundWater buffalo is an economically important livestock species and about half of its total world population exists in India. Development of stem cell technology in buffalo can find application in targeted genetic modification of this species. Testis has emerged as a source of pluripotent stem cells in mice and human; however, not much information is available in buffalo.Objectives and MethodsPou5f1 (Oct 3/4) is a transcription factor expressed by pluripotent stem cells. Therefore, in the present study, expression of POU5F1 transcript and protein was examined in testes of both young and adult buffaloes by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical analysis. Further, using the testis transplantation assay, a functional assay for spermatogonial stem cells (SSCs), stem cell potential of gonocytes/spermatogonia isolated from prepubertal buffalo testis was also determined.ResultsExpression of POU5F1transcript and protein was detected in prepubertal and adult buffalo testes. Western blot analysis revealed that the POU5F1 protein in the buffalo testis exists in two isoforms; large (∼47 kDa) and small (∼21 kDa). Immunohistochemical analysis revealed that POU5F1 expression in prepubertal buffalo testis was present in gonocytes/spermatogonia and absent from somatic cells. In the adult testis, POU5F1 expression was present primarily in post-meiotic germ cells such as round spermatids, weakly in spermatogonia and spermatocytes, and absent from elongated spermatids. POU5F1 protein expression was seen both in cytoplasm and nuclei of the stained germ cells. Stem cell potential of prepubertal buffalo gonocytes/spermatogonia was confirmed by the presence of colonized DBA-stained cells in the basal membrane of seminiferous tubules of xenotransplanted mice testis.Conclusion/SignificanceThese findings strongly indicate that gonocytes/spermatogonia, isolated for prepubertal buffalo testis can be a potential target for establishing a germ stem cell line that would enable genetic modification of buffaloes.

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Section: Discussionsupporting

confidence: 83%

“…In the present study, expression of POU5F1 transcript was detected both in prepubertal and adult buffalo testis. In a recent study, expression of POU5F1 transcript was reported in buffalo embryonic stem cell (ESC) like cells [47]. However in the same study, it was reported that POU5F1 is expressed as four pseudogenes.…”

Section: Discussionmentioning

confidence: 98%