Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Khan, Shagufta; Suryawanshi, Amol Ratnakar; Ranpura, Sandeep A.; Jadhav, S V; Khole, Vrinda

doi:10.1530/rep-09-0052

Cited by 27 publications

(35 citation statements)

References 75 publications

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“…punctatus during spawning. Among other upregulated genes of spawning phase, rgs4 , alcam and cxcl2 have been associated with gonadal stem cells [66–68] while slc6a1 and hipk1 have been reported in spermatids and spermatozoa [69,70]. Nonetheless, it is difficult to decipher the precise role of these genes in regulation of testicular events attributed to spawning phase in C .…”

Section: Discussionmentioning

confidence: 99%

De novo sequencing and comparative analysis of testicular transcriptome from different reproductive phases in freshwater spotted snakehead Channa punctatus

2017

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The spotted snakehead Channa punctatus is a seasonally breeding teleost widely distributed in the Indian subcontinent and economically important due to high nutritional value. The declining population of C. punctatus prompted us to focus on genetic regulation of its reproduction. The present study carried out de novo testicular transcriptome sequencing during the four reproductive phases and correlated differential expression of transcripts with various testicular events in C. punctatus. The Illumina paired-end sequencing of testicular transcriptome from resting, preparatory, spawning and postspawning phases generated 41.94, 47.51, 61.81 and 44.45 million reads, and 105526, 105169, 122964 and 106544 transcripts, respectively. Transcripts annotated using Rattus norvegicus reference protein sequences and classified under various subcategories of biological process, molecular function and cellular component showed that the majority of the subcategories had highest number of transcripts during spawning phase. In addition, analysis of transcripts exhibiting differential expression during the four phases revealed an appreciable increase in upregulated transcripts of biological processes such as cell proliferation and differentiation, cytoskeleton organization, response to vitamin A, transcription and translation, regulation of angiogenesis and response to hypoxia during spermatogenically active phases. The study also identified significant differential expression of transcripts relevant to spermatogenesis (mgat3, nqo1, hes2, rgs4, cxcl2, alcam, agmat), steroidogenesis (star, tkt, gipc3), cell proliferation (eef1a2, btg3, pif1, myo16, grik3, trim39, plbd1), cytoskeletal organization (espn, wipf3, cd276), sperm development (klhl10, mast1, hspa1a, slc6a1, ros1, foxj1, hipk1), and sperm transport and motility (hint1, muc13). Analysis of functional annotation and differential expression of testicular transcripts depending on reproductive phases of C. punctatus helped in developing a comprehensive understanding on genetic regulation of spermatogenic and steroidogenic events in seasonally breeding teleosts. Our findings provide the basis for future investigation on the precise role of testicular genes in regulation of seasonal reproduction in male teleosts.

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Section: Discussionmentioning

confidence: 99%

De novo sequencing and comparative analysis of testicular transcriptome from different reproductive phases in freshwater spotted snakehead Channa punctatus

2017

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“…Upon silver staining, the major band (∼47 kDa) of fraction III was excised from the gel and subjected to trypsin digestion for protein identification using MALDI-TOF/TOF mass spectrometry as described earlier [25]. The peptides were analyzed in MS as well as MS/MS mode to obtain sequence specific information.…”

Section: Methodsmentioning

confidence: 99%

Prostate Secretory Protein of 94 Amino Acids (PSP94) Binds to Prostatic Acid Phosphatase (PAP) in Human Seminal Plasma

et al. 2013

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Prostate Secretory Protein of 94 amino acids (PSP94) is one of the major proteins present in the human seminal plasma. Though several functions have been predicted for this protein, its exact role either in sperm function or in prostate pathophysiology has not been clearly defined. Attempts to understand the mechanism of action of PSP94 has led to the search for its probable binding partners. This has resulted in the identification of PSP94 binding proteins in plasma and seminal plasma from human. During the chromatographic separation step of proteins from human seminal plasma by reversed phase HPLC, we had observed that in addition to the main fraction of PSP94, other fractions containing higher molecular weight proteins also showed the presence of detectable amounts of PSP94. This prompted us to hypothesize that PSP94 could be present in the seminal plasma complexed with other protein/s of higher molecular weight. One such fraction containing a major protein of ∼47 kDa, on characterization by mass spectrometric analysis, was identified to be Prostatic Acid Phosphatase (PAP). The ability of PAP present in this fraction to bind to PSP94 was demonstrated by affinity chromatography. Co-immunoprecipitation experiments confirmed the presence of PSP94-PAP complex both in the fraction studied and in the fresh seminal plasma. In silico molecular modeling of the PSP94-PAP complex suggests that β-strands 1 and 6 of PSP94 appear to interact with domain 2 of PAP, while β-strands 7 and 10 with domain 1 of PAP. This is the first report which suggests that PSP94 can bind to PAP and the PAP-bound PSP94 is present in human seminal plasma.

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“…3,4 Sperm have proteins that are sperm specific, immunogenic, and accessible to antibodies, and hence immunological intervention with these molecules can lead to infertility. 13 The identified peptide sequences of one of the novel protein, sperm flagella protein 2 (SFP2), was similar to the bullous pemphigoid antigen 1, isoform 1 ⁄ 2 ⁄ 3 ⁄ 4 [bullous pemphigoid antigen (BPA) ⁄ BPAG1 ⁄ Dystonin]. [6][7][8] However, immunization against testicular sperm proteins usually leads to orchitis, endocrine disturbance, and loss of libido.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of Contraceptive Potential of a Novel Epididymal Sperm Protein SFP2 in a Mouse Model

Khan

Jadhav

Suryawanshi

et al. 2011

American Journal of Reproductive Immunology

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Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Cited by 27 publications

References 75 publications

De novo sequencing and comparative analysis of testicular transcriptome from different reproductive phases in freshwater spotted snakehead Channa punctatus

De novo sequencing and comparative analysis of testicular transcriptome from different reproductive phases in freshwater spotted snakehead Channa punctatus

Prostate Secretory Protein of 94 Amino Acids (PSP94) Binds to Prostatic Acid Phosphatase (PAP) in Human Seminal Plasma

Evaluation of Contraceptive Potential of a Novel Epididymal Sperm Protein SFP2 in a Mouse Model

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