siRNA-mediated knockdown of sperm-associated antigen 11a (Spag11a) mRNA in epididymal primary epithelial cells affects proliferation: a transcriptome analyses
Differential expression of a variety of proteins in the four major regions of the epididymis contributes to maturation of spermatozoa and region-specific cellular functions as well. Proliferation of epithelial cells of the epididymis is highly controlled and thus is one of the major reasons for the nonoccurrence of cancers in this organ system. The molecular mechanisms and the contribution of region-specific genes in epithelial cell proliferation are not yet fully understood. In this study, for the first time, we analyzed the role of sperm-associated antigen 11a (Spag11a), a caput-specific beta-defensin–like antimicrobial gene in governing epididymal cell proliferation and global gene expression. siRNA-mediated knockdown of Spag11a mRNA in epididymal primary epithelial cells resulted in increased cell proliferation. Out of the 68,842 genes analyzed, 4182 genes were differentially expressed (2154 upregulated and 2028 downregulated). A variety of genes that participate in different cellular processes and pathways were differentially regulated. Genes that are important for epithelial cell proliferation were found to be differentially regulated and these changes were confirmed by real-time PCR. Overexpression of Spag11a in immortalized rat caput epididymal cells resulted in decreased proliferation capacity. Results of this study indicate that Spag11a plays a crucial role in governing epididymal epithelial cell proliferation.
KeywordsEpididymis Cell proliferation siRNA Transcriptome Sperm-associted antigen 11A
We thank the facilities extended by UGC-SAP, UGC-CAS, DBT-CREBB, DST-PURSE, UGC-UPE-II, and FIST programs at School of Life Sciences, University of Hyderabad. Kumari Sangeeta received the RGNF junior research fellowship from Government of India.
The study was funded by a research grant (EMR/2016/000791) of the Science and Engineering Research Board, Government of India is acknowledged. KS received a RGNF research fellowship from Government of India.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Animal ethics approval
All procedures involving animals were conducted using the guidelines for the care and use of laboratory animals to minimize suffering, and this study was specifically approved by the Institutional Animal Ethics Committee of University of Hyderabad (IAEC/UH/2017/01/SY/P13).
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