THY1 is a surface marker of porcine gonocytes
Yi Zheng A B C D , Ying He A B C D , Junhui An A B C , Jinzhou Qin A B C , Yihan Wang A B C , Yaqing Zhang A B C , Xiue Tian A B C E and Wenxian Zeng A B C E
+ Author Affiliations
- Author Affiliations
A College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
B Shaanxi Center of Stem Cells Engineering and Technology, Yangling, Shaanxi, 712100, China.
C Key Lab for Biotechnology of Agriculture Ministry of China, Yangling, Shaanxi, 712100, China.
D These two authors contributed equally to this work.
E Corresponding authors. Emails: zengwnxian@hotmail.com; txe82@yahoo.com.cn
Reproduction, Fertility and Development 26(4) 533-539 https://doi.org/10.1071/RD13075
Submitted: 23 December 2012 Accepted: 17 March 2013 Published: 20 May 2013
Abstract
Gonocytes are important for the study of spermatogenesis. Identification and isolation of gonocytes has been reported in rodents but not in pigs due to a lack of molecular markers for gonocytes. The objective of this study was to identify THY1 expression in porcine testicular tissue and subsequently utilise THY1 as a marker to isolate and enrich porcine gonocytes from testes of newborn piglets. Immunohistochemical analysis showed that THY1 was expressed in gonocytes. Double-immunofluorescent analysis of THY1 and ZBTB16 indicated that THY1 and ZBTB16 were partially co-localised in gonocytes. Double-immunofluorescent analysis of both THY1 and GATA4 suggested that THY1+ cells were not Sertoli cells. Magnetic-activated cell sorting of THY1+ cells yielded a cell population with an enrichment of UCHL1+ gonocytes 3.4-fold of that of the unsorted testicular cell population. Western blot and quantitative reverse transcription–polymerase chain reaction analyses confirmed that the selected THY1+ fraction had a higher expression of UCHL1 than the unsorted cells. In conclusion, the study demonstrated that THY1 is a surface marker of gonocytes in testes of pre-pubertal boars and could be utilised to identify and isolate porcine gonocytes. The findings will also facilitate culture and manipulation of male germline stem cells.
Additional keywords: culture, pig, spermatogonial stem cell.
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