Activity and Localization of NADH-dependant Oxidoreductase (Diaphorase) in Boar Spermatozoa

Atanassov, B.; Denkova, R; Georgiev, G.

doi:10.1111/j.1439-0272.1987.tb02331.x

Cited by 7 publications

(2 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The oxidative enzyme NADPH‐d is also a sperm‐specific key enzyme for sperm metabolism and plays a role in sperm function through electron transfer from NAD(P)H (the promoter of sperm capacitation) to different acceptors, thereby generating ROS (Gavella et al, ). Previous studies (Atanassov et al, ; Kacheva et al, ) demonstrated that NADPH‐d enzyme activity was localized to the mitochondria in the midpiece of the spermatozoa, and Kacheva et al () suggested that low sperm motility in the buffalo may be due to low NADPH‐d activity.…”

Section: Discussionmentioning

confidence: 98%

Region‐specific localization of NOS isoforms and NADPH‐diaphorase activity in the intratesticular and excurrent duct systems of adult domestic cats (Felis catus)

Lïman

Alan

2016

Microscopy Res & Technique

View full text Add to dashboard Cite

Nitric oxide (NO) is produced by nitric oxide synthases (NOSs) and plays an important role in all levels of reproduction from the brain to the reproductive organs. Recently, it has been discovered that all germ cells and Leydig cells in the cat testis exhibit stage-dependent nuclear and cytoplasmic endothelial (eNOS) and inducible (iNOS)-NOS immunoreactivity and cytoplasmic nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) reactivity. As a continuation of this finding, in this study, cellular localization of NADPH-d and immunolocalization and expression of all three NOS isoforms were investigated in the intratesticular (tubuli recti and rete testis), and excurrent ducts (efferent ductules, epididymal duct and vas deferens) of adult cats using histochemistry, immunohistochemistry and western blotting. NADPH-d activity was found in the midpiece of the spermatozoa tail and epithelial cells of all of ducts, except for nonciliated cells of the efferent ductules. Even though the immunoblotting results revealed similar levels of nNOS, eNOS and iNOS in the caput, corpus and cauda segments of epididymis and the vas deferens, immunostainings showed cell-specific localization in the efferent ductules and region- and cell-specific localization in the epididymal duct. All of three NOS isoforms were immunolocalized to the nuclear membrane and cytoplasm of the epithelial cells in all ducts, but were found in the tail and the cytoplasmic droplets of spermatozoa. These data suggest that NO/NOS activity might be of importance not only for the functions of the intratesticular and excurrent ducts but also for sperm maturation.

show abstract

Section: Discussionmentioning

confidence: 98%

Region‐specific localization of NOS isoforms and NADPH‐diaphorase activity in the intratesticular and excurrent duct systems of adult domestic cats (Felis catus)

Lïman

Alan

2016

Microscopy Res & Technique

View full text Add to dashboard Cite

show abstract

“…Diaphorase controls the ratio between reduced and oxidized forms of NADH; thus it takes part in the sperm energy balance (Atanassov et al 1987). This oxidoreductase is integrated with the mitochondrial respiratory chain and generates superoxide anions, affecting the level of reactive oxygen species (ROS) (Fraczek and Kurpisz 2005).…”

Section: Introductionmentioning

confidence: 99%

Sperm mitochondria diaphorase activity a gene mapping study of recombinant inbred strains of mice

Gołas¹,

Malek²,

Piasecka³

et al. 2010

Int. J. Dev. Biol.

View full text Add to dashboard Cite

In order to study the genetic control of semen quality parameters, we derived a set of recombinant inbred (RI) mice from crosses between two inbred strains, KE and CBA/Kw, which differ significantly in gamete quality and fertility parameters. In this work, we used male mice from the two parental strains and from ten RI strains to map genes controlling quantitative traits such as sperm mitochondrial diaphorase activity, and assessed the correlation between this trait, sperm motility and in vivo fertilization efficiency. We analyzed sperm mitochondrial dehydrogenase (diaphorase) activity (NADH-dependent NBT assay) cytochemically by means of computerized image densitometry and obtained values for four parameters: 1) integrated optical density (IOD) for all pixels of the midpiece, 2) mean optical density (MOD) for the midpiece pixels, 3) length of sperm midpiece and 4) area of sperm midpiece. Polymorphic microsatellite marker profiles were prepared for 20 mouse chromosomes in the ten RI strains. We used Map Manager QTX software to correlate the strain distribution patterns (SDPs) of the four measured parameters with the SDPs of the analyzed markers. Hypothetical genes modifying diaphorase activity were mapped to chromosomal region 19q43-19q47, containing, for example, Poll, Sfxn2, Cyp17a1 and Usmg5 genes. Chromosomal regions 18q44 and 18q49-18q80 also showed correlation with the SDPs of diaphorase activity. Katnal2, Me2 and Stard6 candidate genes were proposed from this region. Diaphorase activity in the mouse sperm midpiece did not correlate with in vivo fertilization efficiency, but was negatively correlated with the linearity and straightness of sperm movement.

show abstract

Development of an Image Analysis System to Monitor the Retention of Residual Cytoplasm by Human Spermatozoa: Correlation With Biochemical Markers of the Cytoplasmic Space, Oxidative Stress, and Sperm Function

Gomez,

Buckingham,

Brindle

et al. 1996

Journal of Andrology

257

View full text Add to dashboard Cite

A method has been developed for quantifying the residual cytoplasm present in the midpiece of human spermatozoa, based upon the imaging of NADH oxidoreductase activity. This procedure used NADH and nitroblue tetrazolium as electron donor and acceptor, respectively, and resulted in the discrete staining of the entire midpiece area, including the residual cytoplasm. Image analysis techniques were then used to generate binary images of the midpiece, from which objective measurements of this cellular domain could be undertaken. Such data were found to be highly correlated with biochemical markers of the cytoplasmic space, such as creatine kinase (CK) and glucose‐6‐phosphate dehydrogenase (G‐6‐PDH), in sperm populations depleted of detectable leukocyte contamination. Morphometric analysis of the sperm midpiece was also found to reflect semen quality in that it predicted the proportion of the ejaculate that would be recovered from the high‐density region of Percoll gradients and was negatively correlated with the movement and morphology of the spermatozoa in semen. Variation in the retention of excess residual cytoplasm was also associated with differences in the functional competence of washed sperm preparations, both within and between ejaculates. Thus, within‐ejaculate comparisons of high‐ and low‐density sperm subpopulations revealed a relative disruption of sperm function in the low‐density fraction. This disruption was associated with the presence of excess residual cytoplasm in the midpiece, high concentrations of cytoplasmic enzymes, and the enhanced‐generation reactive oxygen species (ROS). Functional differences between individual high‐density Percoll preparations were also negatively correlated with the area of the midpiece and the corresponding capacity of the spermatozoa to generate ROS. These findings suggest that one of the factors involved in the etiology of defective sperm function is the incomplete extrusion of germ cell cytoplasm during spermiogenesis as a consequence of which the spermatozoa experience a loss of function associated with the induction of oxidative stress.

show abstract

Activity and Localization of NADH-dependant Oxidoreductase (Diaphorase) in Boar Spermatozoa

Cited by 7 publications

References 8 publications

Region‐specific localization of NOS isoforms and NADPH‐diaphorase activity in the intratesticular and excurrent duct systems of adult domestic cats (Felis catus)

Region‐specific localization of NOS isoforms and NADPH‐diaphorase activity in the intratesticular and excurrent duct systems of adult domestic cats (Felis catus)

Sperm mitochondria diaphorase activity a gene mapping study of recombinant inbred strains of mice

Development of an Image Analysis System to Monitor the Retention of Residual Cytoplasm by Human Spermatozoa: Correlation With Biochemical Markers of the Cytoplasmic Space, Oxidative Stress, and Sperm Function

Contact Info

Product

Resources

About