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Jelly plug dissolution in Discoglossus pictus eggs (Anura) involves peroxidase-like activity and oxidative opening of disulphide bonds

Published online by Cambridge University Press:  26 September 2008

G. Pitari ,
S. Dupré ,
C. Fusco ,
G. Maurizi  and
C. Campanella
G. Pitari
Affiliation:
Università dell'Aquila and Università di Roma 'La Sapienza', Italy
S. Dupré
Affiliation:
Università dell'Aquila and Università di Roma 'La Sapienza', Italy
C. Fusco
Affiliation:
Università dell'Aquila and Università di Roma 'La Sapienza', Italy
G. Maurizi
Affiliation:
Università dell'Aquila and Università di Roma 'La Sapienza', Italy
C. Campanella*
Affiliation:
Università dell'Aquila and Università di Roma 'La Sapienza', Italy
*
C. Campanella, Dipartimento di Biologia Evolutiva e Comparata, Università di Napoli, 80134 Napoli, Italy.
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Summary

In amphibian eggs the formation of a capsular chamber is one of the most striking events occurring either upon oviposition or after fertilisation. In the egg of the anuran Discoglossus pictus a capsular chamber forms following fertilisation or activation; the egg with its vitelline envelope rotates in this chamber according to gravity. Previous work showed that the chamber is the product of plug dissolution. The plug is a lens-shaped jelly coat, typical of Discoglossus, covering only part of the animal hemisphere. Its dissolution is caused by material released from the egg about 15 min after fertilisation through exocytosis of at least two types of vacuoles. Liquefaction of the plug correlates with the reduction of disulphide bonds present in the jelly matrix. In this study we investigated the nature of the substances released from the egg and some changes occurring in the plug during liquefaction. SDS-PAGE showed that the proteic profile of the plug changes dramatically after fertilisation, confirming proteic cleavage in the plug matrix during its dissolution. Through in vitro tests and electrophoretic analysis of the Ringer solution in which the egg exudate was collected, an increase in the activity of the solution was determined in the presence of hydrogen peroxide, and peroxidase activity was depicted in the egg exudate. The presence of free thiol groups and cysteic acid residues (or cysteine sulphinic acid) in the plugs of activated eggs was established, suggesting that during plug dissolution some disulphide bonds are oxidatively opened. This suggests that enzyme(s) with peroxidase activity are released following fertilisation. We surmise that such enzymes are contained in the intraovular vacuoles the exocytosis of which triggers the onset of plug liquefaction. The possible release of hydrogen peroxide from the egg is discussed.

Keywords

Anuran eggsDisulphide bondsPeroxidase activityPlug dissolution
Type
Article
Information
Zygote , Volume 1 , Issue 1 , February 1993 , pp. 61 - 69
Copyright
Copyright © Cambridge University Press 1993

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References

Barja de Quiroga, G., Gil, P. & Alonso-Bedate, M. (1985). Catalase enzymatic activity and electrophoretic patterns in adult amphibians: a comparative study. Comp. Biochem. Physiol. 80B, 853–8.Google Scholar
Bellini, L. & Campanella, C. (1974). Dati istochimici sulle gelatine ovalari e sull'ovidutto in Discoglossus pictus in rapporto alia fecondazione. Boll. Zool. 41, 438.Google Scholar
Campanella, C., Talevi, R., Atripaldi, U. & Quaglia, L. (1986). The cortical endoplasmic reticulum and its possible role in activation of Discoglossus pictus (Anura) eggs. In: Molecular and Cellular Biology of Fertilisation, ed. Hedrick, J.L., pp. 185–203. New York: Plenum Publishing Co.CrossRefGoogle Scholar
Campanella, C., Amore, F., Pitari, G., Fusco, C., Maurizi, G. & Dupré, S. (1992). Post-fertilization changes in Discoglossus pictus (Anura) eggs result in the formation of a capsular chamber where the egg rotates. Int. J. Dev. Biol.(in press)Google Scholar
Carroll, E.J. & Hedrick, J.L. (1974). Hatching in the toad Xenopus laevis: morphological events and evidence for a hatching enzyme. Dev. Biol. 38, 1–13.CrossRefGoogle ScholarPubMed
Carroll, E.J., Wei, S.H., Nagel, G.M. & Ruibal, R. (1991). Structure and Macromolecular composition of the egg and embryo jelly coats of the anuran Lepidobatrachus laevis. Dev. Growth Differ. 33, 37–43.CrossRefGoogle ScholarPubMed
Foerder, C.A. & Shapiro, B.M. (1977). Release of ovoperoxidase from sea urchin eggs hardens the fertilization membrane with tyrosine crosslinks. Proc. Natl. Acad. Sci. USA 74, 4214–18.CrossRefGoogle ScholarPubMed
Foerder, C.A., Klebanoff, B.M. & Shapiro, B.M. (1978). Hydrogen peroxide production, chemiluminescence and the respiratory burst of fertilization: interrelated events in early sea urchin development. Proc. Natl. Acad. Sci. USA 75, 3183–7.CrossRefGoogle ScholarPubMed
Ghiara, G. (1960). Osservazioni sugli strati superficiali e corticali di ovociti e di uova di Discoglossus pictus. Arch. Zool. Ital. 45, 9–92.Google Scholar
Heinecke, J.W. & Shapiro, B.M. (1989). Respiratory burst oxidase of fertilization. Proc. Natl. Acad. Sci. USA 86, 1259–63.CrossRefGoogle ScholarPubMed
Hibbard, H. (1928). Contribution a l'érude de l'ovogénèse chez Discoglossus pictus. Otth. Arch. Biol. 38, 251–326.Google Scholar
Hirs, J.M. (1967). Performic acid oxidation. Methods Enzymol. 11,197–9.CrossRefGoogle Scholar
Kao, M. & Elinson, R. P. (1988). The entire mesodermal mantle behaves as Speman's organizer in dorso-anterior enhanced Xenopus laevis embryos. Dev. Biol. 127, 64–77.CrossRefGoogle Scholar
Kapitany, R.A. & Zebrowski, E.J. (1973). A high resolution PAS stain for polyacrylamide gel electrophoresis. Anal. Biochem. 56, 361–9.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680–5.CrossRefGoogle ScholarPubMed
Merril, C.R., Goldman, D., Sedman, S.A. & Ebert, M.H. (1981). Ultrasensitive stain in polyacrylamide gels shows regional variation in cerebro-spinal fluid proteins. Science 211, 1437–1438.CrossRefGoogle Scholar
Monroy, A. (1965). Chemistry and Physiology of Fertilization. New York: Holt, Reinehart & Winston.Google Scholar
Salthe, S.N. (1963). The egg capsule in the Amphibia. Morphol. 113, 161–71.CrossRefGoogle ScholarPubMed
Somers, C.E. & Shapiro, B.M. (1989). Insights into the molecular mechanisms involved in sea urchin fertilization envelope assembly. Dev. Growth Differ. 31, 1–7.CrossRefGoogle ScholarPubMed
Spencer, M.A. (1969). A new convenient method for estimation of total cystine-cysteine in proteins. Anal. Biochem. 32, 185–7.CrossRefGoogle ScholarPubMed
Talevi, R. & Campanella, C. (1988). Fertilization in Discoglossus pictus. I. Sperm-egg interactions in distinct regions of the dimple and occurrence of a late stage of sperm penetration. Dev. Biol. 130, 524–35.CrossRefGoogle ScholarPubMed
Talevi, R., Dale, B. & Campanella, C. (1985). Fertilization and activation potentials in Discoglossus pictus (Anura) eggs. Dev. Biol. 130, 524–35.CrossRefGoogle Scholar
Wakahara, M. (1989). Specification and establishment of dorso-ventroe polarity in eggs and embryos of Xenopus laevis. Dev. Growth Differ. 31,197–207.CrossRefGoogle ScholarPubMed
Weidman, P.J., Kay, E.S. & Shapiro, B.M. (1985). Assembly of the sea urchin fertilization membrane: isolation of proteoliasin, a calcium-dependent ovoperoxidase binding protein. Cell Biol. 100, 938–46.CrossRefGoogle Scholar