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Dicarbonyl L-xylulose reductase (DCXR), a "moonlighting protein" in the bovine epididymis.

Akintayo A, Légaré C, Sullivan R - PLoS ONE (2015)

Bottom Line: Dicarbonyl L-xylulose reductase is secreted in the epididymis luminal compartment in the soluble fraction and is associated with microvesicular elements named epididymosomes.In spermatozoa, the DCXR protein was found in the cytoplasmic and membranous fractions.These data describe DCXR in the bovine epididymis and reveal that its behavior differs from that found in humans.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche du Centre Hospitalier Universitaire (CHU) de Québec, Département d'Obstétrique, Gynécologie et Reproduction, Université Laval, Faculté de Medicine, Québec, Canada.

ABSTRACT
During maturation and the acquisition of their fertilization potential, male germ cells are subjected to various sequential modifications that occur in the epididymis. Protein addition, reorganization or withdrawal, comprise some of these modifications. Dicarbonyl L-xylulose reductase (DCXR), a multifunctional protein involved in various enzymatic and protein interaction processes in different physiological systems, is one of the proteins added to spermatozoa in the epididymis. DCXR is a well-conserved protein with multiple characteristics including enzymatic activities and mediation of cell-cell interaction. In this study, we characterized the DCXR gene and protein expression in the bovine epididymis. Dicarbonyl L-xylulose reductase mRNA is differentially expressed in the caput, corpus, and cauda epididymide epithelial cells with a higher level observed in the cauda region. Tissue protein expression follows the same pattern as the corresponding mRNA expression with a cytoplasmic and apical distribution in the corpus and cauda epithelial cells, respectively. The protein can also be found with a nuclear localization in cauda epididymidis epithelial cells. Dicarbonyl L-xylulose reductase is secreted in the epididymis luminal compartment in the soluble fraction and is associated with microvesicular elements named epididymosomes. In spermatozoa, the DCXR protein was found in the cytoplasmic and membranous fractions. Expression of the DCXR protein is higher on caput spermatozoa but finally shows a weak detection in semen. These data describe DCXR in the bovine epididymis and reveal that its behavior differs from that found in humans. It seems that, in this model, the DCXR protein might have a questionable involvement in the fertilization process.

No MeSH data available.


Immunohistochemical localization of bovine DCXR protein along the bovine epididymis.Caput (A), corpus (B and B*) and cauda (C) epididymidis using anti-DCXR antiserum (A, B and C). A pre-immune rabbit serum was used as a negative control (B*). Arrows indicate DCXR detected as a brown-red staining. The sections were counterstained in blue with Harris hematoxylin. LU = lumen; EP = epithelium; IT = interstitial tissue.
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pone.0120869.g005: Immunohistochemical localization of bovine DCXR protein along the bovine epididymis.Caput (A), corpus (B and B*) and cauda (C) epididymidis using anti-DCXR antiserum (A, B and C). A pre-immune rabbit serum was used as a negative control (B*). Arrows indicate DCXR detected as a brown-red staining. The sections were counterstained in blue with Harris hematoxylin. LU = lumen; EP = epithelium; IT = interstitial tissue.

Mentions: To characterize the expression pattern of the DCXR protein in the epididymis, the complete DCXR mRNA transcript from the cauda section was used to produce a his-tag conjugated recombinant full DCXR protein in a prokaryotic system, and the purified protein (S2 Fig.) was used to produce specific rabbit antisera against bovine DCXR. The specificity and efficiency of the antisera is shown in S3 Fig. and S4 Fig. Western blot analysis on protein extracts from caput, corpus, and cauda regions of the epididymis, revealed a protein of 32 kDa with the same expression pattern as the DCXR gene, i.e., increasing expression from the caput extract to the cauda (Fig. 4A and B). Immunohistochemical assay revealed that the DCXR protein is located in the epithelial cells of the epididymis and at a weaker level in some blood vessels (data not shown). The protein follows the same distribution pattern as the mRNA expression along the epididymis: low level and diffuse cytoplasmic localization in the caput (Fig. 5A), a strong cytoplasmic and apical expression in the corpus (Fig. 5B), and finally, a high level of expression in the basal and apical subcellular compartment in the cauda epididymal epithelium (Fig. 5C). One can notice a high staining in the apical cells of the caput epididymidis epithelium. This is not observed in the corpus or cauda epididymidis (S5 Fig. and S6 Fig.). The anti-DCXR anti-serum used is highly specific as shown by the absence of staining when a pre-immune serum (Fig. 5B inset) or the anti-DCXR sera pre-absorbed with the recombinant protein was used (data not shown).


Dicarbonyl L-xylulose reductase (DCXR), a "moonlighting protein" in the bovine epididymis.

Akintayo A, Légaré C, Sullivan R - PLoS ONE (2015)

Immunohistochemical localization of bovine DCXR protein along the bovine epididymis.Caput (A), corpus (B and B*) and cauda (C) epididymidis using anti-DCXR antiserum (A, B and C). A pre-immune rabbit serum was used as a negative control (B*). Arrows indicate DCXR detected as a brown-red staining. The sections were counterstained in blue with Harris hematoxylin. LU = lumen; EP = epithelium; IT = interstitial tissue.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4376396&req=5

pone.0120869.g005: Immunohistochemical localization of bovine DCXR protein along the bovine epididymis.Caput (A), corpus (B and B*) and cauda (C) epididymidis using anti-DCXR antiserum (A, B and C). A pre-immune rabbit serum was used as a negative control (B*). Arrows indicate DCXR detected as a brown-red staining. The sections were counterstained in blue with Harris hematoxylin. LU = lumen; EP = epithelium; IT = interstitial tissue.
Mentions: To characterize the expression pattern of the DCXR protein in the epididymis, the complete DCXR mRNA transcript from the cauda section was used to produce a his-tag conjugated recombinant full DCXR protein in a prokaryotic system, and the purified protein (S2 Fig.) was used to produce specific rabbit antisera against bovine DCXR. The specificity and efficiency of the antisera is shown in S3 Fig. and S4 Fig. Western blot analysis on protein extracts from caput, corpus, and cauda regions of the epididymis, revealed a protein of 32 kDa with the same expression pattern as the DCXR gene, i.e., increasing expression from the caput extract to the cauda (Fig. 4A and B). Immunohistochemical assay revealed that the DCXR protein is located in the epithelial cells of the epididymis and at a weaker level in some blood vessels (data not shown). The protein follows the same distribution pattern as the mRNA expression along the epididymis: low level and diffuse cytoplasmic localization in the caput (Fig. 5A), a strong cytoplasmic and apical expression in the corpus (Fig. 5B), and finally, a high level of expression in the basal and apical subcellular compartment in the cauda epididymal epithelium (Fig. 5C). One can notice a high staining in the apical cells of the caput epididymidis epithelium. This is not observed in the corpus or cauda epididymidis (S5 Fig. and S6 Fig.). The anti-DCXR anti-serum used is highly specific as shown by the absence of staining when a pre-immune serum (Fig. 5B inset) or the anti-DCXR sera pre-absorbed with the recombinant protein was used (data not shown).

Bottom Line: Dicarbonyl L-xylulose reductase is secreted in the epididymis luminal compartment in the soluble fraction and is associated with microvesicular elements named epididymosomes.In spermatozoa, the DCXR protein was found in the cytoplasmic and membranous fractions.These data describe DCXR in the bovine epididymis and reveal that its behavior differs from that found in humans.

View Article: PubMed Central - PubMed

Affiliation: Centre de Recherche du Centre Hospitalier Universitaire (CHU) de Québec, Département d'Obstétrique, Gynécologie et Reproduction, Université Laval, Faculté de Medicine, Québec, Canada.

ABSTRACT
During maturation and the acquisition of their fertilization potential, male germ cells are subjected to various sequential modifications that occur in the epididymis. Protein addition, reorganization or withdrawal, comprise some of these modifications. Dicarbonyl L-xylulose reductase (DCXR), a multifunctional protein involved in various enzymatic and protein interaction processes in different physiological systems, is one of the proteins added to spermatozoa in the epididymis. DCXR is a well-conserved protein with multiple characteristics including enzymatic activities and mediation of cell-cell interaction. In this study, we characterized the DCXR gene and protein expression in the bovine epididymis. Dicarbonyl L-xylulose reductase mRNA is differentially expressed in the caput, corpus, and cauda epididymide epithelial cells with a higher level observed in the cauda region. Tissue protein expression follows the same pattern as the corresponding mRNA expression with a cytoplasmic and apical distribution in the corpus and cauda epithelial cells, respectively. The protein can also be found with a nuclear localization in cauda epididymidis epithelial cells. Dicarbonyl L-xylulose reductase is secreted in the epididymis luminal compartment in the soluble fraction and is associated with microvesicular elements named epididymosomes. In spermatozoa, the DCXR protein was found in the cytoplasmic and membranous fractions. Expression of the DCXR protein is higher on caput spermatozoa but finally shows a weak detection in semen. These data describe DCXR in the bovine epididymis and reveal that its behavior differs from that found in humans. It seems that, in this model, the DCXR protein might have a questionable involvement in the fertilization process.

No MeSH data available.