Stage-dependent expression of extra-embryonic tissue-spermatogenesis-homeobox gene 1 (ESX1) protein, a candidate marker for X chromosome-bearing sperm
Yueh-Chiao Yeh A , Vie-Cheng Yang A , Shing-Chung Huang B and Neng-Wen Lo B CA Department of Life Science, Tunghai University, Taichung, Taiwan 40744, ROC.
B Department of Animal Science and Biotechnology, Tunghai University, Taichung, Taiwan 40744, ROC.
C Corresponding author. Email: nlo@mail.thu.edu.tw
Reproduction, Fertility and Development 17(4) 447-455 https://doi.org/10.1071/RD04077
Submitted: 22 July 2004 Accepted: 27 January 2005 Published: 23 March 2005
Abstract
Extra-embryonic tissue-spermatogenesis-homeobox gene 1 (Esx1) encodes an X-linked homeobox protein. Despite the fact that the temporal and spatial mRNA expression pattern of the protein has been studied extensively in the testis, specific localisation of ESX1 in the testis remains to be determined. In the present study, we generated ESX1 antiserum to investigate the stage- and tissue-specific expression of ESX1 in the mouse. Western blotting and immunofluorescent analyses revealed that general localisations of ESX1 were consistent with its RNA expression patterns; that is, it was restricted mainly to the placenta and testis. Immunofluorescent studies demonstrated that ESX1 existed in the testes after 3 weeks of age, coincident with the appearance of round spermatids in the seminiferous tubules. Moreover, ESX1 expression became more abundant in the luminal regions of the seminiferous tubules as the development of round spermatids progressed into spermatozoa. In contrast, reduced expression of ESX1 was observed in experimentally induced cryptorchid testes. The later expression of ESX1 suggests a role in post-meiotic germ cell development. To further understand ESX1 expression in sperm with respect to X chromosome-bearing sperm, we used ESX1 antiserum to immunostain sperm by confocal laser microscopy. Approximately half the sperm population was recognised by the ESX1 antiserum. On the basis of results of the present study, we suggest that ESX1 could be used as a protein marker for X chromosome-bearing sperm.
Extra keywords: protein marker, spermatogenesis, testis.
Acknowledgments
The authors express their thanks to Dr Pei-Cheng Lin for technical advice on the preparation of the experimental cryptorchid mice. The authors also thank Wen-Hong Huang, Shu-Ling Lee and Kuo-Chuan Lee for technical assistance. This research was supported by grant NSC 91–2311-B-029–007 from the National Science Council, Taiwan, ROC, to N.W.L.
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