, Volume 65, Issue 4, pp 239–253 | Cite as

Lipopolysaccharide induces epididymal and testicular antimicrobial gene expression in vitro: insights into the epigenetic regulation of sperm-associated antigen 11e gene

  • Barnali Biswas
  • Suresh Yenugu
Original Paper


Infections of the male reproductive tract lead to infertility, and the molecular mechanisms that operate under these conditions are not well studied. Using epididymal and testicular tissues cultured in vitro, we demonstrate that lipopolysaccharide (LPS) induces the mRNA expression of beta-defensins, Spag11s, and pro-inflammatory cytokines in the rat caput, cauda, and testes. LPS-induced antimicrobial gene expression involved NF-kB activation and decreased levels of histone deacetylase 1 (HDAC1) and DNA methyltransferase (DNMT), all of which possibly allow antimicrobial gene transcription. Inhibition of endogenous HDAC1 and DNMT1 resulted in higher antimicrobial gene expression when compared to the LPS alone treated conditions. Increased trimethylation of histone 3, its binding to the upstream region of Spag11e gene, and demethylation of this region were observed during endotoxin challenge. We demonstrate that antimicrobial gene expression in the male reproductive tract tissues during endotoxin challenge involves NF-kB activation and epigenetic changes.


Epididymis Defensins Sperm-associated antigen 11 Lipopolysaccharide Epigenetic DNA methylation 



We thank Dr. Sanjeev Khosla and Bindu Bhargavi, Center for DNA Fingerprinting and Diagnostics, Hyderabad for their help with DNA methylation analyses. We thank the facilities extended by UGC-SAP, UGC-CAS, DBT-CREBB, and FIST programs at School of Life Sciences, University of Hyderabad. BB received a fellowship from University Grants Commission and Council for Scientific and Industrial Research, Government of India.

Supplementary material

251_2012_674_Fig11_ESM.jpg (52 kb)
Supplementary data 1

Defensin 1 (A) and Spag11c (B) mRNA in the control caput, cauda and testis obtained at 0 h (□) and 9 h (■).Values shown are mean ± S.D. (JPEG 51 kb)

251_2012_674_MOESM1_ESM.tif (67 kb)
High resolution image (TIFF 67.1 kb)
251_2012_674_Fig12_ESM.jpg (160 kb)
Supplementary data 2

Nucleotide sequence flanking the transcription start site of Spag11e gene . The CpG dinucleotides are indicated in red and underlined. The transcription start site (ATG) is indicted in bold and underlined. (JPEG 159 kb)

251_2012_674_MOESM2_ESM.tif (115 kb)
High resolution image (TIFF 114 kb)
251_2012_674_Fig13_ESM.jpg (49 kb)
Supplementary data 3

Methylation status of Spag11e gene in the testis. Location of CpG dinucleotides in the Spag11e gene analyzed by isolating the DNA from testis. (JPEG 48 kb)

251_2012_674_MOESM3_ESM.tif (61 kb)
High resolution image (TIFF 60.8 kb)
251_2012_674_MOESM4_ESM.doc (37 kb)
Supplementary data 4 Primer sequences of the genes analysed. (DOC 37 kb)
251_2012_674_MOESM5_ESM.ppt (155 kb)
Supplementary data 5 Densitometric analyses for the Western blots shown in Fig. 2d to determine phospho P65 and phosphor IkBα protein expression in the epididymis and testis after LPS stimulation. Values shown are mean ± S.D. (PPT 155 kb)
251_2012_674_MOESM6_ESM.ppt (152 kb)
Supplementary data 6 Densitometric analyses for the Western blots shown in Fig. 8 to determine HDAC1 protein expression in the epididymis and testis after LPS stimulation. Values shown are mean ± S.D. (PPT 152 kb)
251_2012_674_MOESM7_ESM.ppt (55 kb)
Supplementary data 7 Densitometric analyses for the Western blots shown in Fig. 9 to determine meH3K36 protein expression in the epididymis and testis after LPS stimulation. Values shown are mean ± S.D. (PPT 55 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  1. 1.Department of Animal SciencesUniversity of Hyderabad, P.O. Central UniversityHyderabadIndia

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