Chemoecology

, Volume 22, Issue 3, pp 199–206 | Cite as

Chemical investigations of defensive steroid sequestration by the Asian snake Rhabdophis tigrinus

  • Deborah A. Hutchinson
  • Alan H. Savitzky
  • Akira Mori
  • Gordon M. Burghardt
  • Jerrold Meinwald
  • Frank C. Schroeder
Research Paper

Abstract

Rhabdophis tigrinus is an Asian natricine snake that possesses unusual defensive glands on the dorsal surface of its neck. These nuchal glands typically contain cardiotonic steroidal toxins known as bufadienolides, which are also abundant in the skin of toads. Feeding experiments demonstrated that toads consumed as prey are the ultimate sources of the bufadienolides in nuchal glands of R. tigrinus. Indeed, snakes on a toad-free Japanese island (Kinkasan, Miyagi Prefecture) lack these compounds in their nuchal glands, confirming that these snakes are unable to synthesize defensive bufadienolides. However, when snakes from Kinkasan are fed toads in the laboratory, they accumulate bufadienolides in their nuchal glands, indicating that they have not lost the ability to sequester defensive compounds from prey. In contrast, R. tigrinus from a toad-rich island (Ishima, Tokushima Prefecture) possess large quantities of bufadienolides, reflecting the abundance of toads from which these compounds can be sequestered. Feeding experiments involving gravid R. tigrinus demonstrated that bufadienolides can be provisioned to offspring so that hatchlings are chemically defended before their first toad meal. Maternal provisioning of bufadienolides can take place through two routes: by deposition in yolk and by diffusion in utero, even late in gestation. We applied bufadienolides to the surface of eggs from Kinkasan and found that the embryos are able to take up these compounds into their nuchal glands, demonstrating the feasibility of uptake across the eggshell. Female R. tigrinus provision bufadienolides to their offspring in direct proportion to their own level of chemical defense. By feeding toad-derived bufotoxins to R. tigrinus hatchlings, we determined that the sequestration of these compounds involves at least three types of modification: hydrolytic cleavage of suberylarginine side chains, hydroxylation, and epimerization.

Keywords

Dietary toxins Toad Bufadienolides Nuchal glands Antipredator defense 

References

  1. Akizawa T, Yasuhara T, Kano R, Nakajima T (1985) Novel polyhydroxylated cardiac steroids in the nuchal glands of the snake, Rhabdophis tigrinus. Biomed Res 6:437–441Google Scholar
  2. Brower LP (1984) Chemical defence in butterflies. In: Vane-Wright RI, Ackery PR (eds) The biology of butterflies. Academic Press, London, pp 109–134Google Scholar
  3. Daly JW, Garaffo HM, Spande TF, Jaramillo C, Rand AS (1994) Dietary source for skin alkaloids of poison frogs (Dendrobatidae)? J Chem Ecol 20:943–955CrossRefGoogle Scholar
  4. Daly JW, Garraffo HM, Spande TF, Clark VC, Ma JM, Ziffer H, Cover JF Jr (2003) Evidence for an enantioselective pumiliotoxin 7-hydroxylase in dendrobatid poison frogs of the genus Dendrobates. Proc Natl Acad Sci USA 100:11092–11097PubMedCrossRefGoogle Scholar
  5. Duffey SS (1980) Sequestration of plant natural products by insects. Annu Rev Entomol 25:447–477CrossRefGoogle Scholar
  6. Dumbacher JP, Spande TF, Daly JW (2000) Batrachotoxin alkaloids from passerine birds: a second toxic bird genus (Ifrita kowaldi) from New Guinea. Proc Natl Acad Sci USA 97:12970–12975PubMedCrossRefGoogle Scholar
  7. Dumbacher JP, Wako A, Derrickson SR, Samuelson A, Spande TF, Daly JW (2004) Melyrid beetles (Choresine): a putative source for the batrachotoxin alkaloids found in poison-dart frogs and toxic passerine birds. Proc Natl Acad Sci USA 101:15857–15860PubMedCrossRefGoogle Scholar
  8. Dumbacher JP, Menon GK, Daly JW (2009) Skin as a toxin storage organ in the endemic New Guinean genus Pitohui. Auk 126:520–530CrossRefGoogle Scholar
  9. Eisner T, Goetz MA, Hill DE, Smedley SR, Meinwald J (1997) Firefly “femmes fatales” acquire defensive steroids (lucibufagins) from their firefly prey. Proc Natl Acad Sci USA 94:9723–9728PubMedCrossRefGoogle Scholar
  10. Erspamer V (1994) Bioactive secretions of the amphibian integument. In: Heatwole H, Barthalmus GT, Heatwole AY (eds) Amphibian biology. Surrey Beatty and Sons, Chipping Norton, Australia, pp 176–350Google Scholar
  11. Hutchinson DA, Mori A, Savitzky AH, Burghardt GM, Wu X, Meinwald J, Schroeder FC (2007) Dietary sequestration of defensive steroids in nuchal glands of the Asian snake Rhabdophis tigrinus. Proc Natl Acad Sci USA 104:2265–2270PubMedCrossRefGoogle Scholar
  12. Hutchinson DA, Savitzky AH, Mori A, Meinwald J, Schroeder FC (2008) Maternal provisioning of sequestered defensive steroids by the Asian snake Rhabdophis tigrinus. Chemoecology 18:181–190CrossRefGoogle Scholar
  13. Kamano Y, Kotake A, Hashima H, Inoue M, Morita H, Takeya K, Itokawa H, Nandachi N, Segawa T, Yukita A, Saitou K, Katsuyama M, Pettit GR (1998) Structure-cytotoxic activity relationship for the toad poison bufadienolides. Bioorg Med Chem 6:1103–1115PubMedCrossRefGoogle Scholar
  14. Mori A, Moriguchi H (1988) Food habits of snakes in Japan: a critical review. Snake 20:98–113Google Scholar
  15. Mori A, Layne D, Burghardt GM (1996) Description and preliminary analysis of antipredator behavior of Rhabdophis tigrinus tigrinus, a colubrid snake with nuchal glands. Jpn J Herpetol 16:94–107Google Scholar
  16. Muller JK, Gross TS, Borgert CJ (2007) Topical dose delivery in the reptilian egg treatment model. Environ Toxicol Chem 26:914–919PubMedCrossRefGoogle Scholar
  17. Nakamura K (1935) On a new integumental poison gland found in the nuchal region of a snake, Natrix tigrina. Mem Coll Sci Kyoto Imp Univ 10B:229–240Google Scholar
  18. Nishida R (2002) Sequestration of defensive substances from plants by Lepidoptera. Annu Rev Entomol 47:57–92PubMedCrossRefGoogle Scholar
  19. Porto AM, Baralle FE, Gros EG (1972) Biosynthesis of bufadienolides in toads III. J Steroid Biochem 3:11–17PubMedCrossRefGoogle Scholar
  20. Saporito RA, Garraffo HM, Donnelly MA, Edwards AL, Longino JT, Daly JW (2004) Formicine ants: an arthropod source for the pumiliotoxin alkaloids of dendrobatid poison frogs. Proc Natl Acad Sci USA 101:8045–8050PubMedCrossRefGoogle Scholar
  21. Saporito RA, Donnelly MA, Norton RA, Garraffo HM, Spande TF, Daly JW (2007) Oribatid mites as a major dietary source for alkaloids in poison frogs. Proc Natl Acad Sci USA 104:8885–8890PubMedCrossRefGoogle Scholar
  22. Smith MA (1938) The nucho-dorsal glands of snakes. Proc Zool Soc Lond B 107:575–583, pl IGoogle Scholar
  23. Urano A, Ishihara K (1987) Biology of toads. Shokabo, TokyoGoogle Scholar
  24. Wibbels T, Bull JJ, Crews D (1991) Synergism between temperature and estradiol: a common pathway in turtle sex determination? J Exp Zool 260:130–134PubMedCrossRefGoogle Scholar
  25. Williams BL, Brodie ED Jr, Brodie ED III (2004) A resistant predator and its toxic prey: persistence of newt toxin leads to poisonous (not venomous) snakes. J Chem Ecol 30:1901–1919PubMedCrossRefGoogle Scholar

Copyright information

© Springer Basel AG 2011

Authors and Affiliations

  • Deborah A. Hutchinson
    • 1
  • Alan H. Savitzky
    • 2
  • Akira Mori
    • 3
  • Gordon M. Burghardt
    • 4
    • 5
  • Jerrold Meinwald
    • 6
  • Frank C. Schroeder
    • 6
    • 7
  1. 1.Department of BiologyCoastal Carolina UniversityConwayUSA
  2. 2.Department of Biological SciencesOld Dominion UniversityNorfolkUSA
  3. 3.Department of Zoology, Graduate School of ScienceKyoto UniversityKyotoJapan
  4. 4.Department of PsychologyUniversity of TennesseeKnoxvilleUSA
  5. 5.Department of Ecology and Evolutionary BiologyUniversity of TennesseeKnoxvilleUSA
  6. 6.Department of Chemistry and Chemical BiologyCornell UniversityIthacaUSA
  7. 7.Boyce Thompson Institute, Cornell UniversityIthacaUSA

Personalised recommendations