Purification and properties of alkaline phosphatase from boar seminal plasma

Iyer, Srividya; Daron, Harlow H.; Aull, John L.

doi:10.1530/jrf.0.0820657

Cited by 5 publications

(8 citation statements)

References 18 publications

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“…To better characterize the plasma enzyme, AP thermoinhibition was carried out at 60 °C, as described by Iyer et al . () with some modifications: the samples were kept at 60 °C for 1′15″, 2′30″, 5′, 10′, then placed in ice and the activity was measured within 30″. The same assay was also performed on the thermostable AP from bovine intestinal mucosa as a control.…”

Section: Methodsmentioning

confidence: 99%

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Alkaline phosphatase in boar sperm function

et al. 2013

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SUMMARYAlkaline phosphatase (AP) catalyses the detachment of phosphate residues from different substrates. Its activity has been demonstrated in seminal plasma and spermatozoa from porcine and other mammalian species; anyway, the role of AP in male reproduction has not been clarified yet and the aim of this study was to determine AP function in boar sperm capacitation and in vitro fertilization (IVF). AP activity was assayed in seminal plasma and in uncapacitated and in vitro capacitated (IVC) spermatozoa; in addition, capacitation was studied in presence of different doses of AP (1.2 and 2.5 IU/mL). The effect of different doses of AP (1.2 and 2.5 IU/mL) on several sperm parameters after IVC (viability, acrosome integrity with FITC-PSA, capacitation status with CTC staining, tyrosine phosphorylation) and on fertilizing ability during IVF were also evaluated. High AP activity was detected in seminal plasma, in particular in sperm-rich fraction; a lower activity was detected in uncapacitated spermatozoa while a significant decrease was evidenced after IVC. Viability was not changed by AP supplementation of the capacitating medium, whereas acrosome integrity and capacitation status were significantly affected by 1.2 and 2.5 doses, with a dose-dependent decrease in acrosome-reacted cells as well as in CTC B pattern displaying cells. As for sperm head protein phosphorylation, a decrease in relative fluorescence was detected in AP 2.5 group, if compared with capacitated one. After IVF, a dose-dependent decrease in penetrated oocytes was recorded, with an increase in monospermic zygote rate. In conclusion, we demonstrated that AP activity decreases under capacitating condition and that addition of AP to spermatozoa during capacitation results in a depression of the capacitating process and IVF. We can infer that AP plays a role in keeping spermatozoa quiescent until they are ejaculated and in modulating the acquisition of the fertilizing ability.

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Section: Methodsmentioning

confidence: 99%

“…In addition, Iyer et al . () reported that two isoforms of AP seem to be present in boar seminal plasma.…”

Section: Introductionmentioning

confidence: 98%

Alkaline phosphatase in boar sperm function

et al. 2013

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“…. , , _.. 1/4 ^ τ · j-' · ι η -j t. Λ The present investigation was concerned with the disammals (1)(2)(3)(4)(5)(6). It is secreted into seminal fluid by the ., .…”

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confidence: 99%

“…. ,,., j j " , c r\ Λ u +u * * tnbution of alkaline phosphatase activity in split ejacepididymis in rabbit and dog (5,6) and by the prostate , r , V ^i-ni-, 4 V "· /< o\ χ* Τ ι u-i * u · ulates of human semen, the response of this alkaline and testis in man (1,3 …”

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confidence: 99%

Alkaline Phosphatase in Human Semen: An Investigation Using Enzyme Inhibitors and Gel Electrophoresis

Lewin¹,

Golan²,

Soffer³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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Summary: Human seminal alkaline phosphatase was investigated with respect to its electrophoretic mobility, heat lability, and susceptibility to inhibition by phenylalanine, tartrate, and homoarginine. Total alkaline phosphatase activity in 30 samples of human semen was measured colorimetrically, using /?-nitrophenylphosphate as substrate. Using linear regression analysis, no significant correlation was found between the enzyme activity and the sperm count, sperm motility, semen volume, and the concentrations of seminal inositol and fructose. The alkaline phosphatase activity was higher in the earlier portion of split ejaculate samples. Sodium DL-tartrate (42mmol/l), which inhibits acid phosphatase, did not inhibit seminal alkaline phosphatase significantly. L-Homoarginine (10 mmol/1), an inhibitor of the liver and bone isoenzymes, inhibited the seminal enzyme (53%), whereas L-phenylalanine (12 mmol/1), a strong inhibitor of placental alkaline phosphate, decraased activity by about 10%. Electrophoresis of semen samples on agarose revealed a braod band which was not sharpened after treatment with neuraminidase. Semen total alkaline phosphatase was essentially totally inactivated by heating at 56 °C for 15 min or 10 min at 65 °C; similar behaviour has been reported for the liver and bone isoenzymes. Electrophoresis after heating did not reveal a residual band of heat-stable placental-like alkaline phosphatase. Semen alkaline phosphatase appears to contain more than one isoenzyme, but placental-like alkaline phosphatase cannot be more than a minor component.

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“…Alkaline phosphatase is present in both seminal plasma and spermatozoa. In seminal plasma, it is present in multiple forms that share similar kinetic properties, and it generally originates from epididymal fluid (Strzezek and Glogowski, 1979;Glogowski and Strzezek, 1980;Frenette et al, 1986;Glogowski, 1988;Iyer et al, 1988;Tang, 1998). Alkaline phosphatase is present in prostasomes (Fabiani and Ronquist, 1995) and is a component of sperm plasma membranes (Soucek and Vary, 1984;Parks et al, 1987).…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Alkaline Phosphatase Activity of Boar Semen by Pentoxifylline, Caffeine, and Theophylline

GLOGOWSKI,

DANFORTH,

CIERESZKO

2002

Journal of Andrology

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Methyl xanthines have been used frequently as additives to sperm suspensions in order to improve sperm characteristics. The mechanism of action on spermatozoa is generally assumed to be inhibition of sperm phosphodiesterase activity, resulting in elevation of complementary adenosine monophosphate levels in spermatozoa. The present study was designed to examine the effect of methyl xanthines (pentoxifylline, caffeine, and theophylline) on another important enzyme system, alkaline phosphatase, in boar seminal plasma and spermatozoa. Inhibition of sperm alkaline phosphatase could be distinguished from that of seminal plasma by a paradoxical stimulation by pentoxifylline at lower pH values in spermatozoa. Among the three methyl xanthines, theophylline exhibited the most dramatic inhibition of alkaline phosphatase activity and substrate inhibition was observed with increasing concentrations. Each methyl xanthine had a different action on alkaline phosphatase activity at lower pH; theophylline showed the highest inhibition, caffeine inhibition was not related to pH, and pentoxifylline did not inhibit alkaline phosphatase of seminal plasma and, in fact, it stimulated its activity (or that of a phosphatase with lower pH optimum) in spermatozoa. These results indicate another possible mechanism of action of methyl xanthines on sperm and are in agreement with data indicating that methyl xanthines are not specific inhibitors of sperm phosphodiesterase, because clearly, they inhibit alkaline phosphatase activity as well.

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Purification and properties of alkaline phosphatase from boar seminal plasma

Cited by 5 publications

References 18 publications

Alkaline phosphatase in boar sperm function

Alkaline phosphatase in boar sperm function

Alkaline Phosphatase in Human Semen: An Investigation Using Enzyme Inhibitors and Gel Electrophoresis

Inhibition of Alkaline Phosphatase Activity of Boar Semen by Pentoxifylline, Caffeine, and Theophylline

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