Medaka pp 365-378 | Cite as

Molecular Evolution of Teleostean Hatching Enzymes and Their Egg Envelope Digestion Mechanism: An Aspect of Co-Evolution of Protease and Substrate

  • Shigeki Yasumasu
  • Kaori Sano
  • Mari Kawaguchi


At the hatching of medaka embryos, the egg envelope is completely solubilized by two proteases, high choriolytic enzyme (HCE, or choriolysin H) and low choriolytic enzyme (LCE, or choriolysin L). HCE causes the egg envelope to swell, and LCE solubilizes the swollen envelope. Molecular phylogenetic analysis has shown that the hatching enzyme was originally composed of a single enzyme, and during evolution, the hatching system consisting of two types of enzymes was established by duplication and diversification of the genes. We compared the egg envelope digestion mechanism between zebrafish having the single enzyme, ZHE1, and medaka having the two enzymes, HCE and LCE. The digestion manner of ZHE1 was highly homologous to that of HCE with respect to swelling of the egg envelope. The cross-species digestion experiment using enzymes and substrates of both zebrafish and medaka revealed that the substrate specificity of ZHE1 is quite similar to that of HCE, whereas the specificity of LCE is different from those of ZHE1 and HCE. Further analysis showed that ZHE1 and HCE maintain the character of an ancestral hatching enzyme, swelling of the egg envelope, and that LCE acquires a new function, the complete digestion of the HCE-swollen egg envelope. Considering several factors such as the origin of egg envelope protein and the spawning environment of eggs, we discuss a co-evolutionary aspect of the hatching enzyme and egg envelope.


Black Rockfish Hatch Enzyme Zona Pellucida Domain Hatch Gland Cell High Choriolytic Enzyme 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Bond JS, Beynon RJ (1995) The astacin family of metalloendopeptidases. Protein Sci 4:1247–1261PubMedCrossRefGoogle Scholar
  2. Chang YS, Wang SC, Tsao CC, Huang FL (1996) Molecular cloning, structural analysis, and expression of carp ZP3 gene. Mol Reprod Dev 44:295–304PubMedCrossRefGoogle Scholar
  3. Darie CC, Janssen WG, Litscher ES, Wassarman PM (2008) Purified trout egg vitelline envelope proteins VEβ and VEγ polymerize into homomeric fibrils from dimers in vitro. Biochim Biophysica Acta 1784:385–392CrossRefGoogle Scholar
  4. Hamazaki TS, Nagahama Y, Iuchi I, Yamagami K (1989) A glycoprotein from the liver constitutes the inner layer of the egg envelope (zona pellucida interna) of the fish, Oryzias latipes. Dev Biol 133:101–110PubMedCrossRefGoogle Scholar
  5. Inohaya K, Yasumasu S, Ishimaru M, Ohyama A, Iuchi I, Yamagami K (1995) Temporal and spatial patterns of gene expression for the hatching enzyme in the teleost embryo, Oryzias latipes. Dev Biol 171:374–385PubMedCrossRefGoogle Scholar
  6. Ishida J (1944a) Hatching enzyme in the fresh-water fish, Oryzias latipes. Annot Zool Jpn 22:137–154Google Scholar
  7. Ishida J (1944b) Further studies on the hatching enzyme of the fresh-water fish, Oryzias latipes. Annot Zool Jpn 22:155–164Google Scholar
  8. Jovine L, Qi H, Williams Z, Litscher E, Wassarman PM (2002) The ZP domain is a conserved module for polymerization of extracellular proteins. Nat Cell Biol 4:457–461PubMedCrossRefGoogle Scholar
  9. Jovine L, Darie CC, Litscher ES, Wassarman PM (2005) Zona pellucida domain proteins. Annu Rev Biochem 74:83–114PubMedCrossRefGoogle Scholar
  10. Kawaguchi M, Yasumasu S, Hiroi J, Naruse K, Inoue M, Iuchi I (2006) Evolution of teleostean hatching enzyme genes and their paralogous genes. Dev Genes Evol 216:769–784PubMedCrossRefGoogle Scholar
  11. Kawaguchi M, Yasumasu S, Hiroi J, Naruse K, Suzuki T, Iuchi I (2007) Analysis of the exon–intron structures of fish, amphibian, bird and mammalian hatching enzyme genes, with special reference to the intron loss evolution of hatching enzyme genes in Teleostei. Gene (Amst) 392:77–88CrossRefGoogle Scholar
  12. Kawaguchi M, Nakagawa M, Noda T, Yoshizaki N, Hiroi J, Nishida M, Iuchi I, Yasumasu S (2008) Hatching enzyme of ovoviviparous black rockfish Sebastes schlegelii: environmental adaptation of hatching enzyme and evolutionary aspects of formation of pseudogene. FEBS J 275:2884–2898PubMedCrossRefGoogle Scholar
  13. Kawaguchi M, Fujita H, Yoshizaki N, Hiroi J, Okouchi H, Nagakura Y, Noda T, Watanabe S, Katayama S, Iwamuro S, Nishida M, Iuchi I, Yasumasu S (2009) Different hatching strategies in embryos of two species, pacific herring Clupea pallasii and Japanese anchovy Engraulis japonicus, that belong to the same order Clupeiformes, and their environmental adaptation. J Exp Zool B312:95–107CrossRefGoogle Scholar
  14. Kawaguchi M, Hiroi J, Miya M, Nishida M, Iuchi I, Yasumasu S (2010) Phylogeny-dependent intron–loss evolution of hatching enzyme genes in teleostei. BMC Evol Biol 10:260Google Scholar
  15. Kudo N, Yasumasu S, Iuchi I, Tanokura M (2004) Crystallization and preliminary X-ray analysis of HCE-1, a hatching enzyme of medaka fish, Oryzias latipes. Acta Crystallogr D60:725–726Google Scholar
  16. Kunz YW (2004) Developmental biology of teleost fishes. Springer Netherlands, AmsterdamGoogle Scholar
  17. Lee KS, Yasumasu S, Nomura K, Iuchi I (1994) HCE, a constituent of the hatching enzymes of Oryzias latipes embryos, releases unique proline-rich polypeptides from its natural substrate, the hardened chorion. FEBS Lett 339:281–284PubMedCrossRefGoogle Scholar
  18. Litscher ES, Janssen WG, Darie CC, Wassarman PM (2008) Purified mouse egg zona pellucida glycoproteins polymerize into homomeric fibrils under non-denaturing conditions. J Cell Physiol 214:153–157PubMedCrossRefGoogle Scholar
  19. Llorca O, Trujillo A, Blanco FJ, Bernabeu C (2007) Structural model of human endoglin, a transmembrane receptor responsible for hereditary hemorrhagic telangiectasia. J Mol Biol 365:694–705PubMedCrossRefGoogle Scholar
  20. Lyons CE, Payette KL, Price JL, Huang RCC (1993) Expression and structural analysis of a teleost homologue of a mammalian zona pellucida gene. J Biol Chem 268:21351–21358PubMedGoogle Scholar
  21. Monné M, Han L, Schwend T, Burendahl S, Jovine L (2008) Crystal structure of the ZP-N domain of ZP3 reveals the core fold of animal egg coats. Nature (Lond) 456:653–657CrossRefGoogle Scholar
  22. Murata K, Sasaki T, Yasumasu S, Iuchi I, Enami J, Yasumasu I, Yamagami K (1995) Cloning of cDNAs for the precursor protein of a low-molecular-weight subunit of the inner layer of the egg envelope (chorion) of the fish Oryzias latipes. Dev Biol 167:9–17PubMedCrossRefGoogle Scholar
  23. Murata K, Sugiyama H, Yasumasu S, Iuchi I, Yasumasu I, Yamagami K (1997) Cloning of cDNA and estrogen-induced hepatic gene expression for choriogenin H, a precursor protein of the fish egg envelope (chorion). Proc Natl Acad Sci USA 94:2050–2055PubMedCrossRefGoogle Scholar
  24. Ohi Y, Ogawa N (1970) Electrophoretic fractionation of the hatching enzyme of medaka, Oryzias latipes. Zool Mag (Tokyo) 79:17–18Google Scholar
  25. Okada A, Nagata K, Sano K, Yasumasu S, Kubota K, Ohtsuka J, Iuchi I, Tanokura M (2009) Crystallization and preliminary X-ray analysis of ZHE1, a hatching enzyme from the zebrafish Danio rerio. Acta Crystallogr F65:1018–1020Google Scholar
  26. Qi H, Williams Z, Wassarman PM (2002) Secretion and assembly of zona pellucida glycoproteins by growing mouse oocytes microinjected with epitope-tagged cDNAs for mZP2 and mZP3. Mol Biol Cell 13:530–541PubMedCrossRefGoogle Scholar
  27. Rankin T, Dean J (2000) The zona pellucida: using molecular genetics to study the mammalian egg coat. Rev Reprod 5:114–121PubMedCrossRefGoogle Scholar
  28. Sano K, Inohaya K, Kawaguchi K, Yoshizaki N, Iuchi N, Yasumasu S (2008) Purification and characterization of zebrafish hatching enzyme: an evolutionary aspect of the mechanism of egg envelope digestion. FEBS J 275:5934–5946PubMedCrossRefGoogle Scholar
  29. Sano K, Kawaguchi M, Yoshikawa M, Iuchi I, Yasumasu S (2010) Evolution of the teleostean zona pellucida gene inferred from the egg envelope protein genes of the Japanese eel, Anguilla japonica. FEBS J 277:4674–4684PubMedCrossRefGoogle Scholar
  30. Shoots AFM, Sackers MRJ, Overkamp PSG, Denucé JM (1983) Hatching in the teleost Oryzias latipes: limited proteolysis causes egg envelope swelling. J Exp Zool 226:93–100CrossRefGoogle Scholar
  31. Stehr CM, Hawkes JM (1979) The comparative ultrastructure of egg membrane and associated pore structures in the starry flounder, Platichthys stellatus (Pallas), and pink salmon, Oncorhynchus gorbuscha (Wallbaum). Cell Tissue Res 202:347–356PubMedCrossRefGoogle Scholar
  32. Sugiyama H, Iuchi I (2000) Molecular structure and hardening of egg envelope in fish. Recent Res Dev Comp Biochem Physiol 1:139–161Google Scholar
  33. Sugiyama H, Yasumasu S, Murata K, Iuchi I, Yamagami K (1998) The third egg envelope subunit in fish: cDNA cloning and analysis, and gene expression. Dev Growth Differ 40:35–45PubMedCrossRefGoogle Scholar
  34. Wang H, Gong Z (1999) Characterization of two zebrafish cDNA clones encoding egg envelope protein ZP2 and ZP3. Biochim Biophys Acta 1446:156–160PubMedCrossRefGoogle Scholar
  35. Yamagami K (1972) Isolation of a choriolytic enzyme (hatching enzyme) of the teleost, Oryzias latipes. Dev Biol 29:343–348PubMedCrossRefGoogle Scholar
  36. Yamagami K (1973) Some enzymological properties of a hatching enzyme (chorionase) isolated from the fresh-water teleost, Oryzias latipes. Comp Biochem Physiol 46B:603–616Google Scholar
  37. Yamagami K, Hamazaki TS, Yasumasu S, Masuda K, Iuchi I (1992) Molecular and cellular basis of formation, hardening, and breakdown of the egg envelope in fish. Int Rev Cytol 136:51–92PubMedCrossRefGoogle Scholar
  38. Yasumasu S, Iuchi I, Yamagami K (1988) Medaka hatching enzyme consists of two kinds of proteases which act cooperatively. Zool Sci 5:191–195Google Scholar
  39. Yasumasu S, Iuchi I, Yamagami K (1989a) Purification and partial characterization of high choriolytic enzyme (HCE), a component of the hatching enzyme of the teleost, Oryzias latipes. J Biochem 105:204–211PubMedGoogle Scholar
  40. Yasumasu S, Iuchi I, Yamagami K (1989b) Isolation and some properties of low choriolytic enzyme (LCE), a component of the hatching enzyme of the teleost, Oryzias latipes. J Biochem 105:212–218PubMedGoogle Scholar
  41. Yasumasu S, Yamada K, Akasaka K, Mitsunaga K, Iuchi I, Shimada H, Yamagami K (1992) Isolation of cDNAs for LCE and HCE, two constituent proteases of the hatching enzyme of Oryzias latipes, and concurrent expression of their mRNAs during development. Dev Biol 153:250–258PubMedCrossRefGoogle Scholar
  42. Yasumasu S, Iuchi I, Yamagami K (1994) cDNAs and the genes of HCE and LCE, two constituents of the medaka hatching enzyme. Dev Growth Differ 36:241–250CrossRefGoogle Scholar
  43. Yasumasu S, Kawaguchi M, Ouchi S, Sano S, Murata K, Sugiyama H, Akema T, Iuchi I (2010) Mechanism of egg envelope digestion by hatching enzymes, HCE and LCE in medaka, Oryzias latipes. J Biochem 148:439–448PubMedGoogle Scholar

Copyright information

© Springer 2011

Authors and Affiliations

  1. 1.Department of Materials and Life Sciences, Faculty of Science and TechnologySophia UniversityChiyoda-kuJapan
  2. 2.Graduate Program of Biological ScienceGraduate School of Science and Technology, Sophia UniversityChiyoda-kuJapan
  3. 3.Atmosphere and Ocean Research InstituteThe University of TokyoKashiwaJapan

Personalised recommendations