Extraction and quantification of acrosin, β-N-acetylglucosaminidase, and arylsulfatase-A from equine ejaculated spermatozoa

Brandon, C.I.; Srivastava, Pranay; Heusner, Gary L.; Fayrer-Hosken, R.A.

doi:10.1002/(sici)1097-010x(19971015)279:3<301::aid-jez11>3.0.co;2-c

Cited by 22 publications

(19 citation statements)

References 47 publications

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“…HR [12] and AH [14] are required for sperm to penetrate through the oocyte cumulus cell matrix, and these two enzymes are assumed to be required for penetration of the zona pellucida [2,3], in addition to AS. AS activity in non-capacitated stallion sperm has been reported to be very low after ejaculation, but its activity is very high in capacitated sperm after incubation [2]. In the present study, the activities of HR, AS and AH in dog sperm increased markedly after 7 hr of incubation, compared with 0.5 hr, and the percentages of capacitated sperm after 7 hr of incubation were significantly higher than after 0.5 hr.…”

supporting

confidence: 93%

“…Changes in hyaluronidase, acrosin and N-acetylhexosaminidase activities (mean ± S.E.) in spermatozoa collected from 12 beagle dogs and in canine capacitation medium (CCM) after 0.5 and 7 hr incubation Hyaluronidase Acrosin N-acetylhexosaminidase (p mol/min/mg or ml) (n mol/min/mg or ml) (p mol/min/mg or ml) 0.5 hr 7 hr 0.5 hr 7 hr 0.5 hr 7 hr It is well known that hyaluronidase (HR) [1,3] and acrosin (AS) [4,9] located within sperm acrosomes are very important enzymes for fertilization, and it has also been reported that N-acetylhexosaminidase (AH) in sperm is associated with penetration of the oocyte [2,14]. However, since there have been few reports on these enzymes in dog sperm [5], the authors examined the activities of HR, AS and AH in incubated dog sperm and in the culture medium.…”

mentioning

confidence: 99%

“…The activities of HR, AS and AH were significantly higher after 7 hr of incubation than after 0.5 hr (P<0.05, 0.01) ( Table 1). After 7 hr of incubation all three enzyme activities were detected in the medium following removal of the sperm ( [14] are required for sperm to penetrate through the oocyte cumulus cell matrix, and these two enzymes are assumed to be required for penetration of the zona pellucida [2,3], in addition to AS. AS activity in non-capacitated stallion sperm has been reported to be very low after ejaculation, but its activity is very high in capacitated sperm after incubation [2].…”

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confidence: 99%

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Changes in Hyaluronidase, Acrosin, and N-Acetylhexosaminidase Activities of Dog Sperm after Incubation.

Kawakami

Arai

Oishi

1999

The Journal of Veterinary Medical Science

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ABSTRACT. Hyaluronidase, acrosin and N-acetylhexosaminidase activities were examined in sperm collected from 12 beagle dogs and in culture medium after 0.5 hr and 7 hr of sperm incubation. The activities of the three enzymes were significantly higher at 7 hr than at 0.5 hr (P<0.05, 0.01), and the increases were associated with sperm capacitation. It was considered that the three enzymes in the dog sperm are related to fertilization by reason of the findings of the release of these enzymes from the sperm into the medium after 7 hr of incubation.-KEY WORDS: canine sperm, capacitation, enzyme.J. Vet. Med. Sci. 61(2): 183-184, 1999 The mean (±S.E.) values of %HA and %AR after 7 hr of sperm incubation were 56.0 ± 4.6% and 28.3 ± 1%, respectively (Fig. 1). The activities of HR, AS and AH were significantly higher after 7 hr of incubation than after 0.5 hr (P<0.05, 0.01) ( Table 1). After 7 hr of incubation all three enzyme activities were detected in the medium following removal of the sperm ( [14] are required for sperm to penetrate through the oocyte cumulus cell matrix, and these two enzymes are assumed to be required for penetration of the zona pellucida [2,3], in addition to AS. AS activity in non-capacitated stallion sperm has been reported to be very low after ejaculation, but its activity is very high in capacitated sperm after incubation [2]. In the present study, the activities of HR, AS and AH in dog sperm increased markedly after 7 hr of incubation, compared with 0.5 hr, and the percentages of capacitated sperm after 7 hr of incubation were significantly higher than after 0.5 hr. Thus it was concluded that the three enzyme activities increased in association with sperm capacitation.HR, AS and AH have been found to be released from the head of hamster [14] and human [13] sperm not only during the acrosome reaction but before it. The results of the present study do not demonstrate the release of the three enzymes from acrosome-intact sperm in the dog. However, release of these enzymes from dog sperm into the medium was observed. The acrosome reaction of dog sperm has Table 1. Changes in hyaluronidase, acrosin and N-acetylhexosaminidase activities (mean ± S.E.) in spermatozoa collected from 12 beagle dogs and in canine capacitation medium (CCM) after 0.5 and 7 hr incubation Hyaluronidase Acrosin N-acetylhexosaminidase (p mol/min/mg or ml) (n mol/min/mg or ml) (p mol/min/mg or ml) 0.5 hr 7 hr 0.5 hr 7 hr 0.5 hr 7 hr It is well known that hyaluronidase (HR) [1,3] and acrosin (AS) [4,9] located within sperm acrosomes are very important enzymes for fertilization, and it has also been reported that N-acetylhexosaminidase (AH) in sperm is associated with penetration of the oocyte [2,14]. However, since there have been few reports on these enzymes in dog sperm [5], the authors examined the activities of HR, AS and AH in incubated dog sperm and in the culture medium.The quality of the semen collected by digital manipulation from 12 normal beagle dogs, age of 2-6 years, was evaluated, and the sperm were washed wit...

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supporting

confidence: 93%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Changes in Hyaluronidase, Acrosin, and N-Acetylhexosaminidase Activities of Dog Sperm after Incubation.

Kawakami

Arai

Oishi

1999

The Journal of Veterinary Medical Science

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“…We have previously shown the presence of ASA (68 kDa) on the convex ridge surface of the mouse AI sperm head (Tantibhedhyangkul et al 2002). However, in other mammals ASA has also been shown as an acrosomal enzyme (Dudkiewicz 1984, Nikolajczyk & O'Rand 1992, Brandon et al 1997. Therefore, we asked whether mouse ASA was present as part of the acrosomal content (AC) and on the surface of AR sperm.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, the second aim of our study was to determine whether Cys69, Lys123, and Lys302 in the active site pocket of ASA were important for ASA-ZP interaction. In addition, we wanted to verify whether ASA existed in the acrosome of mouse sperm, as documented in other mammalian sperm (Dudkiewicz 1984, Nikolajczyk & O'Rand 1992, Brandon et al 1997. Information of the ASA localization on the sperm head should shed more light to its roles during sperm binding and penetration through the ZP.…”

Section: Introductionmentioning

confidence: 99%

Sperm arylsulfatase A binds to mZP2 and mZP3 glycoproteins in a nonenzymatic manner

Xu¹,

Liu²,

Srakaew³

et al. 2012

Reproduction

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We have shown previously that sperm surface arylsulfatase A (ASA) of mouse, pig, and human is involved in sperm-egg zona pellucida (ZP) binding. By treating capacitated mouse sperm with A23187 to induce the acrosome reaction, we demonstrated by immunoblotting that ASA also existed in the acrosomal content and on the inner acrosomal membrane. Since mZP2 and mZP3 are known as sperm receptors, whereas mZP1 as a cross-linker of mZP2/mZP3, we determined whether purified ASA bound to mZP2 and mZP3 selectively. The three mZP glycoproteins were purified from solubilized ovarian ZP by size exclusion column chromatography. Immuno-dot blot analyses revealed that purified sperm ASA bound to mZP2 at the highest level followed by mZP3, whereas the binding of ASA to mZP1 was minimal. The results confirmed the physiological significance of sperm ASA in the ZP binding process. The binding of ASA to mZP2 and mZP3 was, however, not dependent on the active site pocket amino acids, Cys69, Lys123, and Lys302, which are pertinent to the capturing of an arylsulfate substrate, since ASA mutant with Ala substitution at these three residues still bound to mZP2 and mZP3. The availability of the active site pocket of ASA bound to the ZP suggested that ASA would still retain enzymatic activity, which might be important for subsequent sperm penetration through the ZP.

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The mammalian acrosome as a secretory lysosome: New and old evidence

Moreno

Alvarado

2006

Molecular Reproduction Devel

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The morphological and biochemical characteristics of the acrosome depart well from any other vesicles in somatic cells, making it one of a kind amongst secretory vesicles. The components of the acrosome include a mixture of unique enzymes like acrosin and other enzymes that when present in somatic cells are commonly found in lysosomes, peroxisomes, and even in the cytoplasm. Several observations have pointed out that acrosomal biogenesis has unique features not previously described in secretory vesicle biogenesis of somatic cells. In this review we discuss the evidence supporting a molecular link between the machinery involved in lysosome and acrosome biogenesis, link which may help account for the acrosome unique composition.

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Extraction and quantification of acrosin, β-N-acetylglucosaminidase, and arylsulfatase-A from equine ejaculated spermatozoa

Cited by 22 publications

References 47 publications

Changes in Hyaluronidase, Acrosin, and N-Acetylhexosaminidase Activities of Dog Sperm after Incubation.

Changes in Hyaluronidase, Acrosin, and N-Acetylhexosaminidase Activities of Dog Sperm after Incubation.

Sperm arylsulfatase A binds to mZP2 and mZP3 glycoproteins in a nonenzymatic manner

The mammalian acrosome as a secretory lysosome: New and old evidence

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