Comparative study on the acrosin activity in sperm of the marine crab Charybdis japonica (A. Milne-Edwards, 1861) (Brachyura, Portunidae) under different environmental conditions

in Crustaceana
Restricted Access
Get Access to Full Text
Rent on DeepDyve

Have an Access Token?

Enter your access token to activate and access content online.

Please login and go to your personal user account to enter your access token.


Have Institutional Access?

Access content through your institution. Any other coaching guidance?


The optimal environmental conditions for acrosin activity in sperm of Charybdis japonica were assessed by orthogonal experiments, and the variation of the acrosin activity in sperm was analysed under hypothermic preservation at 4°C and cryopreservation in liquid nitrogen at −196°C, respectively. The acrosin activity and protein component in sperm from the female spermatheca and from the male seminal receptacle, were also compared. The highest acrosin activity was obtained at pH 8, 25°C and 30‰ salinity. The acrosin activity and viability rates of sperm decreased with the preservation time elapsed at 4°C, and a positive correlation between acrosin activity and viability rates (r=0.987; P<0.05) was recorded. These two indices decreased significantly before and after cryopreservation at −196°C, and subsequently changed slowly in the liquid nitrogen. The cryoprotectant had a significant effect on the viability rates of sperm, but not much on the acrosin activity. The acrosin activity of sperm from the male seminal receptacle was (122.53 ± 1.66) × 10−6 μIU under optimal environmental conditions, which was significantly higher than that of the sperm from the female spermatheca (105.65 ± 2.91) × 10−6 μIU (P<0.01). Three kinds of protein subunits in sperm acrosin from the male seminal receptacle were observed by dissociating SDS-PAGE (71.7, 69.2 and 67.3 kDa), which were degraded to some degree in the female spermatheca (70.4, 66.7 and 64.9 kDa). Several special components were noted in the female spermatheca, with one of higher molecular mass and five of lower molecular mass, which may play an important role in preserving the sperm in the female spermatheca.


International Journal of Crustacean Research



AnchordoguyT.CroweJ. H.GriffinF. J.ClarkW. H., 1988. Cryopreservation of sperm from the marine shrimp Sicyonia ingentis. Cryobiology, 25: 238-243.

BabaT.KashiwabaraS.WatanabeK.ItohH.MichikawaY.KimuraK.TakadaM.FukamizuA.AraiY., 1989. Activation and maturation mechanisms of boar acrosin zymogen based on the deduced primary structure. J. Biol. Chem., 264: 11920-11927.

BrayW. A.LawrenceA. L., 1998. Male viability determinations in Penaeus vannameri: evaluation of short-term storage of spermatophores up to 36 h and comparison of Ca2+-free saline and seawater as sperm homogenate media. Aquaculture, 160: 63-67.

CiereszkoA.DabrowskiK.MimsS. D.GlogowskiJ., 2000. Characteristics of sperm acrosin-like activity of paddlefish (Polyodon spathula Walbaum). Comp. Biochem. Physiol. (B), 125: 197-203.

CuiY. H.ZhaoR. L.WangQ.ZhangZ. Y., 2000. Determination of sperm activity for evaluation of male fertility. Asia. J. Androl., 2: 229-232.

DuP.QiuY. C., 2007. Research progress on the effect factors of the acrosin activity. Chinese J. Andro., 21: 54-57.

FritzH.TschescheH.FinkE., 1976. Proteinase inhibitors from boar seminal plasma. Methods Enzymol., 45: 834-847.

GowJ. C., 2000. Cryopreservation of aquatic invertebrate semen: a review, 2000. Aquaculture Research, 31: 259-271.

GuanW. B.WangG. Z.LiS. J., 2002. Cryopreservation of spermatozoa of mud crab (Scylla serrata) and viability assay by biostain method. J. Oceanogr. Taiwan Stra., 2: 457-463.

GuanW. B.WangG. Z.LiS. J.ChenJ. M., 2005. Acrosin activity of spermatozoa in spermathecae of Scylla serrata. Mar. Sci. Bull., 24: 87-91.

HowesL.JonesR., 2002. Interactions between zona pellucida glycoproteins and sperm proacrosin/acrosin during fertilization. J. Reprod. Immunol., 53: 181-192.

KennedyW. P.KaminskiJ. M.van der VenH. H., 1989. A simple, clinical assay to evaluate the acrosine activity of human spermatozoa. J. Andro., 10: 221-231.

LeungT. J. R.LawrenceA. L., 1987. Observation on the decline in sperm quality of Penaeus setiferus under laboratory conditions. Aquac., 65: 363-370.

LinJ. X.YanP.GuoX.ZengY. F.YaoY. N., 2007. Effects of crypreservation on enzyme activities of wild yak sperm. China Herbivor., 27: 10-13.

LiuH. J.FengL., 2002. Cultivation technique of marine crabs: 246-325. (China Agriculture Press, Beijing).

LiuQ.YangX. Z.ChengY. X.WuX. G.FanL. P.HuangZ. F., 2009. Comparison of sperm morphology acrosin activity and immune indices between precocious and normal Eriocheir sinensis. J. Fish. Sci., 16: 183-191.

LiuR. Z.GuoH.SunY.XueB. G., 2004. Effects of freezing on acrosin activities of human spermatozoa. Chinese J. Laborat. Diag., 8: 647-648.

MazurP., 1977. The role of intraceallular freezing in the death of cells cooled at supraoptimal rates. Cryobiology, 14: 251-272.

PiaoH. X.JinY.HanM. M., 2010. Effects of temperature and storage time on the pig sperm acrosin activity. J. Anhui Agric. Univ., 37: 54-57.

ReichartM.LedermanH.Har-EvenD.KedemP.BartoovB., 1993. Human sperm acrosin activity with relation to semen parameters and acrosomal ultrastructure. Andrology, 25: 59-66.

SharmaR.HoggJ.BromhamD. R., 1993. Is spermatozoan acrosin a predictor of fertilization and embryo quality in the human. Fertil. Steri., 60: 881-887.

WangL.LiC. Y., 1999. Spermathecal ultrastructure of freshwater crab Sinopotamon yantsekiense. Acta Hydrob. Sin., 23: 245-248.

WangZ. S.ZuoW. J.WangC. Z.XuW. G., 1995. Study on assaying acrosin activity of human spermatozoa. J. Norm. Bethune Univ., 21: 465-468.

XuX. H.YanB. L.XuJ. T.XuG. C.PuY. F.WuJ. X.WuC. L., 2010a. Spermatozoa morphology and influence of environmental factors on viability rates in crab Charybdis japonica. Acta Ocean. Sin., 32: 93-99.

XuX. H.YanB. L.XuJ. T.XuG. C.PuY. F.XuJ. H., 2010b. Cryopreservation of spermatozoa in Japanese swimming crab Charybdis japonica. Fisher. Sci., 29: 601-604.

XuX. H.YanB. L.XuJ. T.XuG. C.ShaoY. Z.WuJ. X.DaiJ., 2010c. Morphological changes and induced conditions of acrosome reaction in vitro induction of sperm from marine crab (Charybdis japonica). J. Fisher. China, 34: 1821-1828.

YuC. G.SongH. T.YaoG. Z., 2005. The quantity distribution and biological property of Charybdis japonica in the East China Sea. J. Shanghai Fish. Univ., 14: 40-45.


  • Acrosin activities and viability rates of spermatozoa of Charybdis japonica (A. Milne-Edwards, 1861) under cryopreservation (4°C). The superscript notations a-d denote significant differences (P<0.05) of the index changing with time elapsed. This figure is published in colour in the online edition of this journal, which can be accessed via

    View in gallery
  • Analysis of proteins in acrosin of spermatozoa of Charybdis japonica (A. Milne-Edwards, 1861) by SDS-PAGE. 1, Acrosin of spermatozoa from male seminal receptacle; 2, acrosin of spermatozoa from female spermatheca.

    View in gallery


Content Metrics

Content Metrics

All Time Past Year Past 30 Days
Abstract Views 7 7 4
Full Text Views 0 0 0
PDF Downloads 0 0 0
EPUB Downloads 0 0 0