Marine Biology

, Volume 147, Issue 3, pp 681–697 | Cite as

Correlating gene expression with larval competence, and the effect of age and parentage on metamorphosis in the tropical abalone Haliotis asinina

  • Daniel J. Jackson
  • Nathan Ellemor
  • Bernard M. Degnan
Research Article


The non-geniculate crustose coralline alga (CCA) Mastophora pacifica can induce the metamorphosis of competent Haliotis asinina (Vetigastropoda) larvae. The ability to respond to this natural cue varies considerably with larval age, with a higher proportion of older larvae (e.g. 90 h) able to metamorphose in response to M. pacifica than younger larvae (e.g. 66 h). Here we document the variation in time to acquisition of competence within a larval age class. For example, after 18 h of exposure to M. pacifica, approximately 15 and 36% of 84 and 90-h-old H. asinina larvae had initiated metamorphosis, respectively. This age-dependent response to M. pacifica is also observed when different aged larvae are exposed to CCA for varying periods. A higher proportion of older larvae require shorter periods of exposure to CCA than younger larvae in order to initiate metamorphosis. In this experiment, as in the previous, a small proportion of young larvae were able to respond to brief periods of CCA exposure, suggesting that they had developed the same state of competency as the majority of their older counterparts. Comparisons of the proportions of larvae undergoing metamorphosis between families reveals that parentage also has a significant (P<0.05) affect on whether an individual will initiate metamorphosis at a given age. These familial differences are more pronounced when younger, largely pre-competent larvae (i.e. 66 h old) are exposed to M. pacifica, with proportions of larvae undergoing metamorphosis differing by as much as 10 fold between families. As these data suggest that variation in the rate of development of the competent state has a genetic basis, and as a first step towards identifying the molecular basis to this variation, we have identified numerous genes that are differentially expressed later in larval development using a differential display approach. Spatial expression analysis of these genes suggests that they may be directly involved in the acquisition of competence, or may play a functional role in the postlarva following metamorphosis.



We are grateful to J. Pechenik, S. Degnan and two anonymous reviewers for critical comments that greatly improved this manuscript, and to the staff of Heron Island Research for their assistance with collection and maintenance of abalone broodstock. The Bribie Island Aquaculture Research Centre provided tanks and seawater systems with which experiments were conducted. This research was supported by grants from the Australian Research Council to BMD.


  1. Arndt A, Smith MJ (1998) Genetic diversity and population structure in two species of sea cucumber: differing patterns according to mode of development. Mol Ecol 7:1053–1064CrossRefGoogle Scholar
  2. Arnold JM, Eri R, Degnan BM, Lavin MF (1997a) Novel gene containing multiple EGF-like motifs transiently expressed in the papillae of the ascidian tadpole larva. Dev Dyn 210:264–273Google Scholar
  3. Arnold JM, Kennett C, Degnan BM, Lavin MF (1997b) Transient expression of a novel serine protease in the ectoderm of the ascidian Herdmania momus during development. Dev Gen Evol 206:455–463Google Scholar
  4. Ayre DJ, Hughes TP (2000) Genotypic diversity and gene flow in brooding and spawning corals along the Great Barrier Reef, Australia. Evolution 54:1590–1605PubMedGoogle Scholar
  5. Baxter GT, Morse DE (1992) Cilia from abalone larvae contain a receptor-dependent G protein transduction system similar to that in mammals. Biol Bull 183:147–154Google Scholar
  6. Bishop CD, Brandhorst BP (2001) NO/cGMP signaling and HSP90 activity represses metamorphosis in the sea urchin Lytechinus pictus. Biol Bull 201:394–404Google Scholar
  7. Bishop CD, Bates WR, Brandhorst BP (2001) Regulation of metamorphosis in ascidians involves NO/cGMP signaling and HSP90. J Exp Zool 289:374–384Google Scholar
  8. Bonar DB (1978) Morphogenesis at metamorphosis in opisthobranch molluscs. In: Chia F-S, Rice ME (eds) Settlement and metamorphosis of marine invertebrate larvae. Elsevier, New York, pp 177–196Google Scholar
  9. Breton S, Dufresne F, Desrosiers G, Blier PU (2003) Population structure of two northern hemisphere polychaetes, Neanthes virens and Hediste diversicolor (Nereididae), with different life-history traits. Mar Biol 142:707–715Google Scholar
  10. Burke RD (1983) The induction of metamorphosis of marine invertebrate larvae. Stimulus and response. Can J Zool 61:1701–1719Google Scholar
  11. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159CrossRefPubMedGoogle Scholar
  12. Clare AS, Thomas RF, Rittschof D (1995) Evidence for the involvement of cyclic AMP in the pheremonal modulation of barnacle settlement. J Exp Biol 198:655–664Google Scholar
  13. Collin R (2001) The effects of mode of development on phylogeography and population structure of North Atlantic Crepidula (Gastropoda: Calyptraeidae). Mol Ecol 10:2249–2262CrossRefPubMedGoogle Scholar
  14. Coon SL, Bonar DB, Weiner RM (1985) Induction of settlement and metamorphosis of the Pacific oyster, Crassostrea gigas (Thunberg), by L-DOPA and catecholamines. J Exp Mar Biol Ecol 94:211–221Google Scholar
  15. Counihan R, McNamara DC, Souter DC, Jebreen EJ, Preston NP, Johnson CR, Degnan BM (2001) Pattern, synchrony and predictability of spawning of the tropical abalone Haliotis asinina from Heron Reef, Australia. Marine Ecology Progress Series 213:193–202Google Scholar
  16. Degnan BM, Morse DE (1995) Developmental and morphogenetic gene regulation in Haliotis rufescens larvae at metamorphosis. Am Zool 35:391–398Google Scholar
  17. Degnan BM, Groppe JC, Morse DE (1995) Chymotrypsin mRNA expression in digestive gland amoebocytes: cell specification occurs prior to metamorphosis and gut morphogenesis in the gastropod, Haliotis rufescens. Roux’s Arch Dev Biol 205:97–101Google Scholar
  18. Degnan BM, Souter D, Degnan SM, Long SC (1997) Induction of metamorphosis in larvae of the ascidian Herdmania momus with potassium ions requires attainment of competence and an anterior signalling center. Dev Gen Evol 206:370–376Google Scholar
  19. Edmands S, Potts DC (1997) Population genetic structure in brooding sea anemones (Epiactis spp.) with contrasting reproductive modes. Mar Biol 127:485–498CrossRefGoogle Scholar
  20. Eri R, Arnold JM, Hinman VF, Green KM, Jones MK, Degnan BM, Lavin MF (1999) Hemps, a novel EGF-like protein, plays a central role in ascidian metamorphosis. Development 126:5809–5818Google Scholar
  21. Giusti AF, Hinman VF, Degnan SM, Degnan BM, Morse DE (2000) Expression of a Scr/Hox5 gene in the larval central nervous system of the gastropod Haliotis, a non-segmented spiralian lophotrochozoan. Evol Dev 2:294–302Google Scholar
  22. Hadfield MG (1977) Chemical interactions in larval settling of a marine gastropod. In: Faulkner DJ, Fenical WH (eds) Marine natural products chemistry. Plenum, San Diego, pp 403–413Google Scholar
  23. Hadfield MG (1978) Metamorphosis in marine molluscan larvae: an analysis of stimuls and response. In: Chia FS, Rice ME (eds) Settlement and metamorphosis of marine invertebrate larvae. Elsevier, Amsterdam, pp 165–175Google Scholar
  24. Hadfield MG (1984) Settlement requirements of molluscan larvae: new data on chemical and genetic roles. Aquaculture 39:283–298Google Scholar
  25. Hadfield MG (1998) The DP Wilson lecture. Research on settlement and metamorphosis of marine invertebrate larvae: past, present and future. Biofouling 12:9-29Google Scholar
  26. Hadfield MG (2000) Why and how marine-invertebrate larvae metamorphose so fast. Cell Dev Biol 11:437–443Google Scholar
  27. Hadfield MG, Strathmann MF (1996) Variability, flexibility and plasticity in life histories of marine invertebrates. Oceanol Acta 19:323–334Google Scholar
  28. Hellberg ME (1996) Dependence of gene flow on geographic distance in two solitary corals with different larval dispersal capabilities. Evolution 50:1167–1175Google Scholar
  29. Hoskin MG (1997) Effects of contrasting modes of larval development on the genetic structures of populations of three species of prosobranch gastropods. Mar Biol 127:647–656Google Scholar
  30. Jackson DJ, Leys SP, Hinman VF, Woods R, Lavin MF, Degnan BM (2002) Ecological regulation of development: induction of marine invertebrate metamorphosis. Int J Dev Biol 46:679–686Google Scholar
  31. Knight J, Rowley AF, Yamazaki M, Clare AS (2000) Eicosanoids are modulators of larval settlement in the barnacle, Balanus amphitrite. J Mar Biol Assoc UK 80:113–117Google Scholar
  32. Kyle CJ, Boulding EG (2000) Comparative population genetic structure of marine gastropods (Littorina spp.) with and without pelagic larval dispersal. Marine Biology 137:835–845Google Scholar
  33. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132PubMedGoogle Scholar
  34. Lee Y-H, Huang GM, Cameron RA, Graham G, Davidson EH, Hood L, Britten RJ (1999) EST analysis of gene expression in early cleavage-stage urchin embryos. Development 126:3857–3867Google Scholar
  35. Lehninger AL, Nelson DL, Cox MM (1993) Principles of biochemistry. Worth, New YorkGoogle Scholar
  36. Marshall DJ, Keough MJ (2003a) Sources of variation in larval quality for free-spawning marine invertebrates: egg size and the local sperm environment. Invertebr Reprod Dev 44:63–70Google Scholar
  37. Marshall DJ, Keough MJ (2003b) Variation in the dispersal potential of non-feeding invertebrate larvae: the desperate larva hypothesis and larval size. Mar Ecol Prog Ser 255:145–153Google Scholar
  38. Marshall DJ, Bolton TF, Keough MJ (2003) Offspring size affects the post-metamorphic performance of a colonial marine invertebrate. Ecology 84:3131–3137Google Scholar
  39. Martin KJ, Pardee AB (1999) Principles of Differential Display. Methods Enzymol 303:234–258Google Scholar
  40. Morse DE, Hooker N, Duncan H, Jensen L (1979) γ-aminobutyric acid, a neurotransmitter, induces planktonic abalone larvae to settle and begin metamorphosis. Science 204:407–410Google Scholar
  41. Moss GA (1999) Factors affecting settlement and early post-settlement survival of the New Zealand abalone Haliotis australis. N Z J Mar Freshw Res 33:271–278Google Scholar
  42. Moss GA, Tong LJ (1992a) Effect of stage of larval development on the settlement of the abalone, Haliotis iris. N Z J Mar Freshw Res 26:69–73Google Scholar
  43. Moss GA, Tong LJ (1992b) Techniques for enhancing larval settlement of the abalone, Haliotis iris, on artificial surfaces. N Z J Mar Freshw Res 26:75–79Google Scholar
  44. O’Brien E, Degnan BM (2002a) Developmental expression of a class IV POU gene in the gastropod Haliotis asinina supports a conserved role in sensory cell development in bilaterians. Dev Gen Evol 212:394–398Google Scholar
  45. O’Brien E, Degnan BM (2002b) Pleiotropic developmental expression of HasPOUIII, a class III POU gene, in the gastropod Haliotis asinina. Mech Dev 114:129–202Google Scholar
  46. Pasquinelli AE, Reinhart BJ, Slack F, Martindale MQ, Kuroda MI, Maller B, Hayward DC, Ball EE, Degnan B, Muller P, Spring J, Srinivasan A, Fishman M, Finnerty J, Corbo J, Levine M, Leahy P, Davidson E, Ruvkun G (2000) Conservation of the sequence and temporal expression of let-7 heterochronic regulatory RNA. Nature 408:86–89Google Scholar
  47. Pechenik JA, Gee CC (1993) Onset of Metamorphic Competence in Larvae of the Gastropod Crepidula Fornicata (L.), Judged by a Natural and an Artificial Cue. J Exp Mar Biol Ecol 167:59–72Google Scholar
  48. Pechenik JA, Heyman WD (1987) Using KCl to determine size at competence for larvae of the marine gastropod Crepidula Fornicata (L.). J Exp Mar Biol Ecol 112:27–38Google Scholar
  49. Pechenik JA, Qian PY (1998) Onset and maintenance of metamorphic competence in the marine polychaete Hydroides elegans Haswell in response to three chemical cues. J Exp Mar Biol Ecol 226:51–74Google Scholar
  50. Pechenik JA, Li W, Cochrane DE (2002) Timing is everything: the effects of putative dopamine antagonists on metamorphosis vary with larval age and experimental duration in the prosobranch gastropod Crepidula fornicata. Biol Bull 202:137–147Google Scholar
  51. Raimondi PT, Keough MJ (1990) Behavioural variability in marine larvae. Aust J Ecol 15:427–437Google Scholar
  52. Roberts R (2001) A review of settlement cues for larval abalone (Haliotis spp.). J Shellfish Res 20:571–586Google Scholar
  53. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  54. Spaulding DC, Morse DE (1991) Purification and characterization of sulfatases from Haliotis rufescens: evidence for changes in synthesis and heterogeneity during development. J Comp Physiol 161:498–515Google Scholar
  55. Strathmann R (1974) The spread of sibling larvae of sedentary marine invertebrates. Am Nat 108:29–44Google Scholar
  56. Todd CD, Lambert WJ, Thorpe JP (1998) The genetic structure of intertidal populations of two species of nudibranch molluscs with planktotrophic and pelagic lecithotrophic larval stages: are pelagic larvae “for” dispersal? J Expl Mar Biol Ecol 228:1-28Google Scholar
  57. Trapido-Rosenthal HG (1986) Initial characterization of receptors for molecules that induce the settlement and metamorphosis of Haliotis rufescens larvae. Diss Abstr Int B 47Google Scholar
  58. Trapido-Rosenthal HG, Morse DE (1986a) Availability of chemosensory receptors is down regulated by habituation of larvae to a morphogenetic signal. Proc Nat Acad Sci 83:7658–7662Google Scholar
  59. Trapido-Rosenthal HG, Morse DE (1986b) Regulation of receptor-mediated settlement and metamorphosis in larvae of a gastropod mollusc (Haliotis rufescens). Bull Mar Sci 39:383–392Google Scholar
  60. Wodicka LM, Morse DE (1991) cDNA sequences reveal mRNAs for two G[alpha] signal transducing proteins from larval cilia. Biol Bull 180:318–327Google Scholar
  61. Woods RG, Roper KE, Gauthier M, Bebell LM, Sung K, Degnan BM, Lavin MF (2004) Gene expression during early ascidian metamorphosis requires signalling by Hemps, an EGF-like protein. Development 131:2921–2933Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Daniel J. Jackson
    • 1
  • Nathan Ellemor
    • 1
  • Bernard M. Degnan
    • 1
  1. 1.School of Integrative BiologyUniversity of QueenslandBrisbaneAustralia

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