Rabbit Epididymal Secretory Proteins. II. Immunolocalization and Sperm Association of REP381

Nixon, Brett; Jones, Russell C.; Clarke, Hannah G; Holland, Michael

doi:10.1095/biolreprod67.1.140

Cited by 6 publications

(6 citation statements)

References 35 publications

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“…For this purpose, aliquots of QEP containing w100 mg protein were precipitated as described previously (Nixon et al 2002a). To each sample, 1.25 ml Pharmalytes (GE Healthcare, Piscataway, NJ, USA), 1.5 ml Destreak (GE Healthcare), and trace amounts of bromophenol blue were added and the samples were used to rehydrate immobilized pH gradient (IPG) gel strips (pH 3-10, 7 cm, nonlinear IPG gels; GE Healthcare) overnight.…”

Section: Sds-pagementioning

confidence: 99%

Post-testicular sperm maturation and identification of an epididymal protein in the Japanese quail (Coturnix coturnix japonica)

Nixon

Ewen²,

Krivanek³

et al. 2014

Reproduction

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The role of the avian epididymis in post-testicular development and capacitation was examined to assess whether avian spermatozoa undergo any processes similar to those characteristic of mammalian sperm development. We found no evidence of a need for quail sperm to undergo capacitation and 90% of testicular sperm could bind to a perivitelline membrane and acrosome react. However, computer-assisted sperm analysis showed that 20% of testicular sperm from the quail were capable of movement and only about 12% of the motile sperm would have a curvilinear velocity greater than the mean for sperm from the distal epididymis. Nevertheless, epididymal transit was associated with increases in mean sperm velocity and the proportion of motile sperm. Together, these findings explain why earlier workers have achieved some fertilizations following inseminations of testicular spermatozoa and also demonstrate the need for some epididymal maturation of avian spermatozoa. Analysis of the electrophoretic profile of quail epididymal luminal proteins revealed that only one major protein (w16 kDa) is secreted by the epididymis and it was virtually the only protein secreted by the ipsilateral epididymis following unilateral orchidectomy. Mass spectrometry showed that this protein is hemoglobin; this finding was confirmed using anti-hemoglobin antibodies. It is suggested that hemoglobin may support sperm metabolism in the quail epididymis, aid in motility, and/or serve as an antioxidant.

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Section: Sds-pagementioning

confidence: 99%

Post-testicular sperm maturation and identification of an epididymal protein in the Japanese quail (Coturnix coturnix japonica)

Nixon

Ewen²,

Krivanek³

et al. 2014

Reproduction

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Section: Isolation Of Rep52 and Preparation Of Polyclonal Antibodiesmentioning

confidence: 99%

“…REP52 was purified from the epididymal fluid of region 8 by preparative SDS-PAGE and electroelution according to the methods described by Nixon et al [4]. Protein purity was assessed on silver-stained gels, and the crude protein preparations were then used to generate antibodies in female BALB/c mice as previously described [4]. Immune sera were collected from the orbital sinus of individual mice and purified over a Hi-Trap Protein G sepharose column (Pharmacia Biotech, Uppsala, Sweden), and the specificity of the purified IgG was analyzed by immunoblotting.…”

Section: Isolation Of Rep52 and Preparation Of Polyclonal Antibodiesmentioning

confidence: 99%

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Molecular and Functional Characterization of the Rabbit Epididymal Secretory Protein 52, REP521

Nixon

Jones

Holland

2008

Biology of Reproduction

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As part of a systematic study of rabbit epididymal proteins involved in sperm maturation, we have identified and characterized a novel glycoprotein (rabbit epididymal secretory protein 52 [REP52]) of 52 kDa. REP52 is synthesized and secreted in a tissue-specific manner by the mid (region 6) and distal (region 7) corpus epididymidis and associates weakly with the sperm surface overlying the principal piece of the tail. Sequencing of cloned REP52 cDNA demonstrated that this protein represents a novel member of the highly conserved fibronectin type II (FN2) module protein family. The protein appears related but not homologous to ungulate seminal plasma proteins and is the first known example to be identified as a rabbit epididymal secretory protein. Consistent with other members of this protein family, REP52 possessed a high level of sequence identity within the FN2 module-encoding domains, but a highly variable N-terminal sequence that failed to show significant homology with published sequences. By analogy with evidence from studies of the ungulate seminal plasma proteins it is hypothesized that the tandemly arranged FN2 modules could facilitate the association of REP52 with sperm phosphatidylcholine residues on the outer leaflet of the sperm tail. It is also considered likely that these domains represent key elements for the function of this novel protein, a conclusion supported by the fact that antisera raised against the REP52 protein blocked in vitro fertilization in a concentration-dependent fashion.

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“…For decades, research has sought to characterize the complex interplay between epididymal secretory/absorptive activity and sperm maturation. A focus of this research has been to document the complement of proteins secreted into the epididymal lumen to characterize their ability to influence sperm maturation (Syntin et al ., ; Nixon et al ., ,b,c; Dacheux et al ., ,b; Dacheux & Dacheux, ; Nixon et al ., , , ; Dacheux et al ., ; Labas et al ., ; Belleannee et al ., ,b; Zhou et al ., ). Taken together, this work has highlighted the importance of specialized epididymal segments as playing discrete roles in the post‐testicular maturation and storage of spermatozoa.…”

Section: Introductionmentioning

confidence: 99%

Profiling of epididymal small non‐protein‐coding RNAs

et al. 2019

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Background: Our understanding of epididymal physiology and function has been transformed over the three decades in which the International Meeting Series on the Epididymis has been hosted. This transformation has occurred along many fronts, but among the most significant advances has been the unexpected discovery of the diversity of small non-protein-coding RNAs (sRNAs) expressed in the epididymal epithelium and differentially accumulated in the luminal population of spermatozoa. Objectives: Here we survey recent literature pertaining to profiling the sRNA landscape of the mammalian epididymis with the goal of demonstrating the contribution that these key regulatory elements, and their associated pathways, make to epididymal physiology and sperm maturation. Results and discussion: High throughput sequencing strategies have fueled an unprecedented advance in our understanding of RNA biology. In the last decade, such high throughput profiling tools have been increasingly applied to study the mammalian epididymis, presaging the discovery of diverse classes of sRNA expressed along the length of the tract. Among the best studied sRNA classes are the microRNAs (miRNA), a sRNA species shown to act in concert with endocrine signals to fine-tune the segmental patterning of epididymal gene expression. In addition to performing this homeostatic role, epithelial cell-derived sRNAs also selectively accumulate into the epididymosomes and spermatozoa that occupy the duct lumen. This exciting discovery alludes to a novel form of intracellular communication that contributes to the establishment of the sperm epigenome and its modification under conditions of paternal stress. Conclusion: Compelling literature has identified sRNAs as a crucial regulatory tier that allows the epididymis to fulfill its combined roles of sperm transport, maturation, and storage. Continued research in this emerging field will contribute to our growing understanding of the etiology of male factor infertility and potentially allow for the future design of rational therapeutic options for these individuals.

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Rabbit Epididymal Secretory Proteins. II. Immunolocalization and Sperm Association of REP381

Cited by 6 publications

References 35 publications

Post-testicular sperm maturation and identification of an epididymal protein in the Japanese quail (Coturnix coturnix japonica)

Post-testicular sperm maturation and identification of an epididymal protein in the Japanese quail (Coturnix coturnix japonica)

Molecular and Functional Characterization of the Rabbit Epididymal Secretory Protein 52, REP521

Profiling of epididymal small non‐protein‐coding RNAs

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