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PMID: 23815807 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Sperm-associated antigen 11A is expressed exclusively in the principal cells of the mouse caput epididymis in an androgen-dependent manner.

Reproductive biology and endocrinology : RB&E ·Vol. 11 ·2013-07-01 ·Pages 59

Pujianto DA, Loanda E, Sari P, Midoen YH, Soeharso P

Abstract

Epididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode the involved proteins remain largely uncharacterized. Previous studies have demonstrated that the genes involved in sperm maturation are regulated by androgen. Spag11a is an epididymal gene that is influenced by androgen. However, little is known about the putative role of this gene in the sperm maturation process. The objective of this study was to characterize Spag11a in the mouse epididymis. In silico analyses were performed to predict signal peptides and functional domains. Spag11a expression was measured by quantitative real-time RT-PCR. Western blots and immunocytochemistry were performed to determine protein expression. SPAG11A is a member of the beta defensin protein family and constitutes a secretory protein. Spag11a was expressed exclusively in the epididymis. Moreover, it exhibited region-specific expression in the caput, which is typical for genes that are involved in creating a suitable microenvironment for sperm maturation. Mouse Spag11a was regulated by androgen. A significant decrease of Spag11a expression was observed at third day following a gonadectomy (P < 0.001). Interestingly, testosterone replacement therapy was able to maintain the expression almost at the normal level, indicating a dependency on androgen. Besides androgen, testicular factors influenced Spag11a expression in a different way. This was revealed by efferent duct ligation in which Spag11a was transiently up-regulated at the third day following the ligation before returning to the normal level at day 5. Spag11a regional expression was also observed at protein level detected by western immunoblotting which revealed a clear band in the caput but not in other regions. The prediction that SPAG11A is a secretory protein was confirmed by immunocytochemical analyses indicating cell-specific expression mainly in the caput principal cells and detection of the protein in epididymal luminal fluid and spermatozoa. Based on the characteristics of Spag11a, it is likely that this gene has a specific role in epididymal sperm maturation. Further studies using functional assays are necessary to confirm this finding.

MeSH Terms
Amino Acid Sequence Androgens/metabolism Animals Base Sequence Binding Sites/genetics Blotting, Western Epididymis/cytology,metabolism Gene Expression Profiling Immunohistochemistry Male Mice Molecular Sequence Data Oligonucleotide Array Sequence Analysis Orchiectomy Phosphorylation Reverse Transcriptase Polymerase Chain Reaction Sperm Maturation/genetics Spermatozoa/metabolism Time Factors beta-Defensins/genetics,metabolism
Chemicals
Androgens Spag11 protein, mouse beta-Defensins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Pujianto Dwi A
Department of Biology, Faculty of Medicine, University of Indonesia, Jl, Salemba Raya 6, Jakarta 10430, Indonesia. [email protected]
Loanda Evelyn
Sari Puji
Midoen Yurnadi H
Soeharso Purnomo
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Article Info
Journal
Reproductive biology and endocrinology : RB&E
Abbr.
Reprod Biol Endocrinol
ISSN
1477-7827
Published
2013-07-01
Epub
2013-00-01
Pages
59
Language
English
Region
England
NLM ID
101153627
PMCID
PMC3710511
Subset
IM
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