Identification of Epididymis‐Specific Antigens Using Neonatal Tolerization

Khole, Vrinda; Joshi, Saurabh; Singh, Sapna

doi:10.1111/j.8755-8920.2000.440606.x

Cited by 11 publications

(22 citation statements)

References 18 publications

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“…Klinefelter & Hamilton (1985) have shown that mammalian sperm on leaving the testis acquire fertilizing ability after interacting with epididymal secretory products especially in the caput region. Report from our earlier study has also indicated that the 29 kDa protein appeared from distal caput and increased in the corpus region (Khole et al 2000). Our present data also indicate similar pattern of reactivity.…”

Section: Discussionsupporting

confidence: 90%

“…The approach used in the present study was modified to overcome the lack of high throughput capacity of the earlier approach. The approach differs in terms of several aspects from the studies reported earlier (Ensrud & Hamilton 1991, Khole et al 2000. First, in this study, we used extracts from different sperm domains, i.e.…”

Section: Discussionmentioning

confidence: 99%

“…SI protocol used earlier (Khole et al 2000) was modified for the present study and is diagrammatically depicted in Fig. 6.…”

Section: Subtractive Immunizationmentioning

confidence: 99%

“…Reactivity of PI sera with rat testis and cauda epididymal sperm proteins extracted in 1% (w/v) SDS was carried out as described earlier (Khole et al 2000). Sera from all animals were diluted 1:100 while secondary antibody Rabbit anti mouse conjugated HRP (Dako) was diluted 1:1000.…”

Section: Soluble Elisamentioning

confidence: 99%

“…Several approaches have been exploited for identification of epididymal proteins such as use of lectins (Srivastav 2000), sera from infertile patients (Primakoff et al 1990, Bohring & Krause 2003, Bhande & Naz 2007, mAbs (Hegde et al 1991), subtractive screening of cDNA library (Kirchhoff 1998), computational biology (Penttinen et al 2003), micro-array (Johnston et al 2005), neonatal tolerization (Ensrud & Hamilton 1991, Khole et al 2000, and proteomics (Syntin et al 1996, Naaby-Hansen et al 1997, Shetty et al 1999, Domagała et al 2007. Proteomics has also been applied to evaluate different aspects of normal sperm function , Ficarro et al 2003, Baker et al 2005).…”

Section: Introductionmentioning

confidence: 99%

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Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Khan

Suryawanshi²,

Ranpura³

et al. 2009

Reproduction

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Functionally immature spermatozoa leave the testis mature during epididymal transit. This process of maturation involves either addition of new proteins or modification of existing proteins onto the sperm domains that are responsible for domain-specific functions. Epididymal proteins are preferred targets for immunocontraception. In an attempt to identify epididymis-specific sperm proteins, we used a novel combinatorial approach comprising subtractive immunization (SI) followed by proteomics. Following SI, sera of mice were used for immunoproteomics, which led to the identification of 30 proteins, of which four proteins namely sperm head protein 1, sperm flagella protein 2 (SFP2), SFP3, and SFP4 are being reported for the first time on sperm. Another group of four proteins namely collagen a-2 (I) chain precursor, homeodomain-interacting protein kinase 1, GTP-binding protein Rab1, and ubiquinol cytochrome c reductase core protein II although reported earlier in testis are being reported for the first time in epididymal sperm. Furthermore, seven out of these eight novel proteins could be validated using peptide ELISA. These data are a useful repository, which could be exploited to develop targets for post-testicular immunocontraception or biomarkers for infertility diagnosis and management.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

“…SI protocol used earlier (Khole et al 2000) was modified for the present study and is diagrammatically depicted in Fig. 6.…”

Section: Subtractive Immunizationmentioning

confidence: 99%

Section: Soluble Elisamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Khan

Suryawanshi²,

Ranpura³

et al. 2009

Reproduction

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Monoclonal Antibodies to Epididymis‐specific Proteins Using Mice Rendered Immune Tolerant to Testicular Proteins

Joshi

Ranpura

Khan

et al. 2003

Journal of Andrology

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Monoclonal antibodies (mabs) have been used as a powerful tool for identification of newer sperm proteins. However, conventional hybridoma technology rarely provides chance to obtain mabs to epididymal proteins. To increase this chance, we have used an alternate method of neonatal tolerization. In this protocol, animals were tolerized at birth using testicular proteins followed by immunization with cauda epididymal sperm protein (which is a cocktail of proteins both from testicular and epididymal origin). This protocol induced a specific immune response to epididymal sperm proteins. Spleen from one of these animals was then used for preparation of mabs. This fusion resulted in a number of mabs reacting specifically to epididymal proteins. Although mabs identified a protein of approximately similar molecular weight on 1-dimensional Western blot analysis, there were differences in regional localization on rat sperm as seen by indirect immunofluorescence. Immunohistochemical localization of these proteins in rat epididymis showed region specific synthesis. The synthesis of proteins was seen in the distal caput epididymis, and maximum expression was seen in supranuclear region of corpus epithelium. The proteins were localized on sperm from corpus and cauda region. Epididymis specific synthesis of the proteins and agglutinating nature of the mabs to these underlines the functional importance of these proteins in sperm maturation in epididymis. These antibodies could therefore, be used as tools for understanding the physiology of maturation of sperm in epididymis and role of the epididymal protein in fertilization.

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Molecular approaches to contraceptive development

Natraj

2001

J Biosci

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The next generation of contraceptives will be based on the identification of novel molecules essential for reproductive processes and will rely on the refinement of older as well as newer technologies. Functional analysis of naturally occurring reproductive genetic disorders and creation of mice null for specific genes would greatly assist in the choice of genetic targets for contraceptive development. Structure-based design of drugs as exemplified by the preparation of an orally active non-peptide gonadotropin releasing hormone (GnRH) would revolutionize drug formulation and delivery for a peptide analogue. This review examines some of the molecular targets that may change contraceptive choices in the future.

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Identification of Epididymis‐Specific Antigens Using Neonatal Tolerization

Abstract: The results of this study demonstrate that neonatal tolerization with testicular sperm proteins, followed by immunization with epididymal sperm proteins, enhances the production of antibodies to proteins exclusively of epididymal origin.

Cited by 11 publications

References 18 publications

Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Identification of novel immunodominant epididymal sperm proteins using combinatorial approach

Monoclonal Antibodies to Epididymis‐specific Proteins Using Mice Rendered Immune Tolerant to Testicular Proteins

Molecular approaches to contraceptive development

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