Secretory mouse quiescin sulfhydryl oxidase 1 aggregates defected human and mouse spermatozoa in vitro and in vivo.

A flavin-dependent enzyme quiescin Q6 sulfhydryl oxidase 1 (QSOX1) catalyzes the oxidation of thiol groups into disulfide bonds. QSOX1 is prominently expressed in the seminal plasma. However, its role in male reproduction is elusive. Here, we purified the secreted form of QSOX1, i.e., QSOX1c, from mouse seminal vesicle secretions and revealed for the first time its function involved in sperm physiology. Exogenous addition of QSOX1c time-dependently promoted the in vitro aggregation of thiol-rich, oxidative stressed, and apoptotic mouse and human sperm cells. Also, in vivo aggregated sperm cells collected from mouse uterine and human ejaculates also showed high levels of QSOX1c, intracellular reactive oxygen species, annexin V, and free thiols. In summary, our studies demonstrated that QSOX1c could agglutinate spermatozoa susceptible to free radical attack and apoptosis. This characteristic may provide an opportunity to separate defective sperm cells and improve sperm quality before artificial insemination in humans and animals.

Point-of-care N-terminal pro B-type natriuretic peptide assay to screen apparently healthy cats for cardiac disease in general practice.

BACKGROUND: Point-of-care (POC) N-terminal pro B-type natriuretic peptide (NT-proBNP) ELISA test has been evaluated for screening cats for cardiac disease in the referral veterinary setting but less is known about its use in general practice (GP). OBJECTIVES: To evaluate the diagnostic utility of a POC NT-proBNP ELISA in cats seen in GPs. ANIMALS: Two hundred and seventeen apparently healthy cats from 21 GPs. METHODS: This was a prospective, cross-sectional study. Cardiac auscultation and POC NT-proBNP ELISA were done by veterinarians at their GPs. After enrollment at GPs, cats were sent to a cardiology referral hospital for cardiac auscultation and echocardiographic diagnosis. Results were interpreted based on whether cats had normal or abnormal echocardiographic findings. RESULTS: Point-of-care NT-proBNP ELISA results differentiated cats in the abnormal group from those in the normal group with a sensitivity of 43%, specificity of 96%. In cats with a heart murmur at GPs, POC NT-proBNP ELISA results differentiated cats in the abnormal group from those in the normal group with a sensitivity of 71% and a specificity of 92%. CONCLUSION AND CLINICAL IMPORTANCE: In apparently healthy cats in GPs, positive POC NT-proBNP results are associated with heart disease, warranting an echocardiogram, but negative results do not reliably exclude heart disease. These results suggest POC NT-proBNP is not an effective screening test for apparently healthy cats in GPs, although its performance is improved if it is used only in cats that have a heart murmur.

Identification, characterization and purification of porcine Quiescin Q6-Sulfydryl Oxidase 2 protein.

BACKGROUND: Post-spermiogenesis membrane surface modifications rely on molecules present in the reproductive tracts. Two isoforms (isoform 1 and 2) from Quiescin Q6-Sulfydryl Oxidase protein family have been identified in the male reproductive tract of rodent species. However, unlike isoform 1, scarce information is available for isoform 2, likely due to its lower expression level and lack of proper purification methods to obtain sufficient protein quantity for further assays. RESULTS: This study demonstrated the presence of short and long forms of Quiescin Q6-Sulfydryl Oxidase 2 in boar, likely representing the secretory (short form) and transmembrane (long form) forms of Quiescin Q6-Sulfydryl Oxidase 2. Immunohistochemistry studies revealed the presence of Quiescin Q6-Sulfydryl Oxidase 2 in a broad range of porcine tissues; the pronounced vesicle-contained Quiescin Q6-Sulfydryl Oxidase 2 at the apical region of epididymis and seminal vesicles epithelium suggested its involvement in sperm physiology and its participation in semen formation. The majority of porcine Quiescin Q6-Sulfydryl Oxidase 2 could be purified via either antibody affinity column or be salted out using 10%-40% ammonium sulfate. Higher amount of low molecular weight Quiescin Q6-Sulfydryl Oxidase 2 observed in the seminal vesicle likely represents the secretory form of Quiescin Q6-Sulfydryl Oxidase 2 and reflects an exuberant secretory activity in this organ. CONCLUSIONS: We demonstrated for the first time, the presence of Quiescin Q6-Sulfydryl Oxidase 2 in porcine species; moreover, two forms of Quiescin Q6-Sulfydryl Oxidase 2 were identified and exhibited distinct molecular weights and properties during protein purification processes. This study also provided feasible Quiescin Q6-Sulfydryl Oxidase 2 purification methods from slaughterhouse materials that could potentially allow obtaining sufficient amount of Quiescin Q6-Sulfydryl Oxidase 2 for future functional investigations.

Roles of the reproductive tract in modifications of the sperm membrane surface.

Successful fertilization requires viable and functional spermatozoa to recognize and fuse with the oocyte. In most mammalian species, mature spermatozoa are not capable of fertilizing the oocytes immediately after ejaculation. However, unlike somatic cells, spermatozoa, after leaving the testis, are transcriptionally and translationally silent; therefore, upon completion of spermiogenesis, spermatozoa carry only a minimal amount of essential proteins on their membranes as well as within their restricted volume of cytoplasm. To develop into a fully functional and competent sperm that is capable of successful fertilization, modifications of the sperm membrane surface during its transit in the reproductive tracts is critical. These post-spermatogenesis modifications advance the maturation of epididymal spermatozoa. In addition, components secreted into the lumen of the reproductive tracts that are later added onto the sperm membrane surface also regulate (inhibit or activate) the functions of the spermatozoa. This acquisition of additional proteins from the reproductive tracts may compensate for the inactivity of morphologically mature spermatozoa. In this review, we discuss the contributions of the male and female genital tracts to modifications of the sperm membrane surface at different stages of fertilization.