Advertisement

Journal of Chemical Ecology

, Volume 31, Issue 11, pp 2501–2518 | Cite as

California Ground Squirrel (Spermophilus beecheyi) Defenses against Rattlesnake Venom Digestive and Hemostatic Toxins

  • James E. Biardi
  • David C. Chien
  • Richard G. Coss
Article

Abstract

Previous studies have shown that some mammals are able to neutralize venom from snake predators. California ground squirrels (Spermophilus beecheyi) show variation among populations in their ability to bind venom and minimize damage from northern Pacific rattlesnakes (Crotalus oreganus), but the venom toxins targeted by resistance have not been investigated. Four California ground squirrel populations, selected for differences in local density or type of rattlesnake predators, were assayed for their ability to neutralize digestive and hemostatic effects of venom from three rattlesnake species. In Douglas ground squirrels (S. b. douglasii), we found that animals from a location where snakes are common showed greater inhibition of venom metalloprotease and hemolytic activity than animals from a location where snakes are rare. Effects on general proteolysis were not different. Douglas ground squirrels also reduced the metalloprotease activity of venom from sympatric northern Pacific rattlesnakes (Crotalus oreganus oreganus) more than the activity of venom from allopatric western diamondback rattlesnakes (C. atrox), but enhanced fibrinolysis of sympatric venom almost 1.8 times above baseline levels. Two Beechey ground squirrel (S. b. beecheyi) populations had similar inhibition of venoms from northern and southern Pacific rattlesnakes (C. o. helleri), despite differences between the populations in the locally prevalent predator. However, the venom toxins inhibited by Beechey squirrels did vary among venom from Pacific rattlesnake subspecies, and between these venoms and venom from allopatric western diamondback rattlesnakes. Blood plasma from Beechey squirrels showed highest inhibition of metalloprotease activity of northern Pacific rattlesnake venom, general proteolytic activity and hemolysis of southern Pacific rattlesnake venom, and hemolysis by allopatric western diamondback venom. These results reveal previously cryptic variation in venom activity against resistant prey that suggests reciprocal adaptation at the molecular level.

Key Words

Spermophilus beecheyi Crotalus oreganus Crotalus atrox venom natural resistance coevolution predator–prey 

Notes

Acknowledgments

This manuscript was greatly improved by the comments of two anonymous reviewers. J.E.B. was supported by a Floyd and Mary Schwall Dissertation Fellowship in Biomedical Research.

References

  1. Anai, K., Suguki, M., Yoshida, E., Maruyama, M. 2002Neutralization of a snake venom hemorrhagic metalloproteinase prevents coagulopathy after subcutaneous injection of Bothrops jararaca venom in ratsToxicon406368CrossRefPubMedGoogle Scholar
  2. Ashton, K. G., Queiroz, A. 2001Molecular systematics of the Western Rattlesnake, Crotalus viridis (Viperidae), with comments on the utility of the D-loop in phylogenetic studies of snakesMol. Phylogenet. Evol.21176189CrossRefPubMedGoogle Scholar
  3. Astrup, T., Müllertz, S. 1952The fibrin plate method for estimating fibrinolytic activityArch. Biochem. Biophys.40346351CrossRefPubMedGoogle Scholar
  4. Baramova, E. N., Shannon, J. D., Bjarnason, J. B., Fox, J. W. 1989Degradation of extracellular matrix proteins by hemorrhagic metalloproteinasesArch. Biochem. Biophys.2756371CrossRefPubMedGoogle Scholar
  5. Bee, A., Theakston, R. D. G., Harrison, R. A., Carter, S. D. 2001Novel in vitro assays for assessing the haemorrhagic activity of snake venoms and for demonstration of venom metalloproteinase inhibitorsToxicon3914291434CrossRefPubMedGoogle Scholar
  6. Biardi, J. E. 2000. Adaptive variation and coevolution in California ground squirrel (Spermophilus beecheyi) and rock squirrel (Spermophilus variegatus) resistance to rattlesnake venom. Ph.D. dissertation, University of California, Davis.Google Scholar
  7. Biardi, J. E., Coss, R. G., Smith, D. G. 2000California ground squirrel (Spermophilus beecheyi) blood sera inhibits crotalid venom proteolytic activityToxicon38713721CrossRefPubMedGoogle Scholar
  8. Black, C. 1963A review of North American tertiary SciuridaeBull. Mus. Comp. Zool. Harv. Univ.130113248Google Scholar
  9. Brattstrom, B. H. 1953Records of Pleistocene reptiles from CaliforniaCopeia1953174179Google Scholar
  10. Braud, S., Bon, C., Wisner, A. 2000Snake venom proteins acting on hemostasisBiochimie82851859CrossRefPubMedGoogle Scholar
  11. Catanese, J. J., Kress, L. F. 1993Opossum serum α1-proteinase inhibitor: Purification, linear sequence, and resistance to inactivation by rattlesnake venom metalloproteinasesBiochemistry32509515CrossRefPubMedGoogle Scholar
  12. Chiszar, D., Walters, A., Urbaniak, J., Smith, M., Mackessy, S. P. 1999Discrimination between envenomated and nonenvenomated prey by Western Diamondback Rattlesnakes (Crotalus atrox): Chemosensory consequences of venomCopeia1999640648Google Scholar
  13. Coss, R. G. 1999Effects of relaxed natural selection on the evolution of behaviorFoster, S. A.Endler, J. A. eds. Geographic Variation in Behavior: Perspectives on Evolutionary MechanismsOxford University PressOxford180208Google Scholar
  14. Coss, R. G., Goldthwaite, R. O. 1995The persistence of old designs for perceptionThompson, N. S. eds. Perspectives in Ethology 11: Behavioral DesignPlenum PressNew York83148Google Scholar
  15. Coss, R. G., Owings, D. H. 1989Rattler battlersNat. Hist.53035Google Scholar
  16. Coss, R. G., Gusé, K. L., Poran, N. S., Smith, D. G. 1993Development of antisnake defenses in California ground squirrels (Spermophilus beecheyi). II. Microevolutionary effects of relaxed selection from rattlesnakesBehaviour124137164Google Scholar
  17. Creer, S., Malhotra, A., Thorpe, R. S., Stocklin, R., Favreau, P., Chou, W. H. 2003Genetic and ecological correlates of intraspecific variation in pitviper venom composition detected using matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) and isoelectric focusingJ. Mol. Evol.56317329CrossRefPubMedGoogle Scholar
  18. Daltry, J. C., Ponnudurai, G., Shin, C. K., Tan, N.-H., Thorpe, R. S., Wüster, W. 1996aElectrophoretic profiles and biological activities: Intraspecific variation in the venom of the Malayan pit viper (Calloselasma rhodostoma)Toxicon346779CrossRefGoogle Scholar
  19. Daltry, J. C., Wüster, W., Thorpe, R. S. 1996bDiet and snake venom evolutionNature379537540CrossRefGoogle Scholar
  20. Fitch, H. S. 1948Ecology of the California ground squirrel on grazing landsAm. Midl. Nat.39513596Google Scholar
  21. Fitch, H. S. 1949Study of snake populations in central CaliforniaAm. Midl. Nat.41513579Google Scholar
  22. Futuyma, D. J., Slatkin, M. 1983CoevolutionSinauer AssociatesSunderland, MAGoogle Scholar
  23. Goldthwaite, R. 1989. Ground squirrel antipredator behavior: Time, chance and divergence. Ph.D. dissertation, University of California, Davis.Google Scholar
  24. Gomulkiewicz, R., Thompson, J. N., Holt, R. D., Nuismer, S. L., Hochberg, M. E. 2000Hot spots, cold spots, and the geographic mosaic theory of coevolutionAm. Nat.156156174CrossRefPubMedGoogle Scholar
  25. Goodwin, R. L., Baumann, H., Berger, F. G. 1996Patterns of divergence during evolution of α-1 proteinase inhibitors in mammalsMol. Biol. Evol.13346358PubMedGoogle Scholar
  26. Grinnell, J., Camp, C. L. 1917A distribution list of amphibians and reptiles of CaliforniaUniv. Calif. Publ. Zool.17127137Google Scholar
  27. Gutiérrez, J. M., Rucavado, A. 2000Snake venom metalloproteinases: Their role in the pathogenesis of local tissue damageBiochimie82841850CrossRefPubMedGoogle Scholar
  28. Hall, E. R. 1981The Mammals of North America Vol. 1WileyNew YorkGoogle Scholar
  29. Heatwole, H., Poran, N. S. 1995Resistances of sympatric and allopatric eels to sea-snake venomsCopeia1136147Google Scholar
  30. Heatwole, H., Powell, J. 1998Resistance of eels (Gymnothorax) to the venom of sea kraits (Laticauda colubrine): A test of coevolutionToxicon36619625CrossRefPubMedGoogle Scholar
  31. Heatwole, H., Poran, N., King, P. 1999Ontogenetic changes in the resistance of bullfrogs (Rana catesbeiana) to the venom of copperheads (Agkistrodon contortrix contortrix) and cottonmouths (Agkistrodon piscivorus piscivorus)Copeia3808814Google Scholar
  32. Hersek, M. J., Owings, D. H. 1993Tail flagging by adult California ground squirrels: A tonic signal that serves different functions for males and femalesAnim. Behav.45129138CrossRefGoogle Scholar
  33. Hill, R. E., Hastie, N. D. 1987Accelerated evolution in the reactive centre regions of serine protease inhibitorsNature3269699CrossRefPubMedGoogle Scholar
  34. Hofmann, H., Bon, C. 1987Blood coagulation induced by the venom of Bothrops atrox. I. Identification, purification and properties of a prothrombin activatorBiochemistry26772780CrossRefPubMedGoogle Scholar
  35. Holman, J. A. 1979A review of North American tertiary snakesPubl. Mus. Mich. State Univ. Paleo. Series1203260Google Scholar
  36. Huang, S.-Y., Pérez, J. C. 1980Comparative study on hemorrhagic and proteolytic activities of snake venomsToxicon18203260Google Scholar
  37. Jorge da Silva, N.,Jr., Aird, S. D. 2001Prey specificity, comparative lethality and compositional differences of coral snake venomsComp. Biochem. Physiol., Part C128425456Google Scholar
  38. Klauber, L. M. 1972Rattlesnakes, Their Habits, Life Histories and Influence on Mankind, Vol. 1University of California PressBerkeleyGoogle Scholar
  39. Kordis, D., Gubensik, F. 2000Adaptive evolution of animal toxin multigene familiesGene2614352CrossRefPubMedGoogle Scholar
  40. Linsdale, J. M. 1946The California Ground Squirrel: A Record of Observations Made on the Hastings Natural History ReservationUniversity of California PressBerkeleyGoogle Scholar
  41. Mackessy, S. P. 1988Venom ontogeny in the Pacific rattlesnakes Crotalus viridis helleri and C. v. oreganusCopeia198892101Google Scholar
  42. Mackessy, S. P. 1993Fibrinogenolytic proteases from the venoms of juvenile and adult northern Pacific rattlesnakes (Crotalus viridis oreganus)Comp. Biochem. Physiol.106B181189Google Scholar
  43. Mackessy, S. P. 1996Characterization of the major metalloprotease isolated from the venom of the northern Pacific rattlesnake, Crotalus viridis oreganusToxicon3412771285CrossRefPubMedGoogle Scholar
  44. Mackessy, S. P., Williams, K., Aston, K. G. 2003Ontogenetic variation in venom composition and diet of Crotalus oreganus concolor: A case of venom paedomorphosis?Copeia4769782Google Scholar
  45. Markland, F. S. 1998Snake venoms and the hemostatic systemToxicon3617491800CrossRefPubMedGoogle Scholar
  46. Miller, L. H. 1912Contributions to avian paleontology from the Pacific coast of North AmericaUniv. Calif. Bull. Dept. Geol.761115Google Scholar
  47. Mishell, B. B., Shiigi, S. M. 1980Selected Methods in Cellular ImmunologyFreemanSan FranciscoGoogle Scholar
  48. Nuismer, S. L., Thomspon, J. N., Gomulkiewicz, R. 1999Gene flow and geographically structured coevolutionProc. R. Soc. Lond., B Bio. Sci.266605609Google Scholar
  49. Nussbaum, R. A., Brodie, E. D.,Jr., Storm, R. M. 1983Amphibians and Reptiles of the Pacific NorthwestUniversity of Idaho PressMoscow, IDGoogle Scholar
  50. Owings, D. H., Coss, R. G. 1977Snake mobbing by California ground squirrels: Adaptive variation and ontogenyBehaviour625069Google Scholar
  51. Owings, D. H., Coss, R. G., McKernon, D., Rowe, M. P., Arrowood, P. C. 2001Snake-directed antipredator behavior of rock squirrels (Spermophilus variegatus): Population differences and snake-species discriminationBehavior138575595CrossRefGoogle Scholar
  52. Ownby, C. L. 1982Pathology of rattlesnake envenomationTu, A. T. eds. Rattlesnake Venoms, Their Actions and TreatmentMarcel DekkerNew York163210Google Scholar
  53. Palmer, M. A. 1993A gelatin test to detect activity and stability of proteases produced by Dichelobacter (bacteriodes) nodosusVet. Microbiol.36113122CrossRefPubMedGoogle Scholar
  54. Perales, J., Domont, G. B. 2002Are inhibitors of metalloproteinases, phospholipases A2 and myotoxins members of the innate immune system?Ménez, X. eds. Perspectives in Molecular ToxinologyWileyNew York435456Google Scholar
  55. Pérez, J. C., Sánchez, E. E. 1999Natural protease inhibitors to hemorrhagins in snake venoms and their potential use in medicineToxicon37703728CrossRefPubMedGoogle Scholar
  56. Pook, C. E., Wüster, W., Thorpe, R. S. 2000Historical biogeography of the Western rattlesnake (Serpentes: Viperidae: Crotalus viridis), inferred from mitochondrial DNA sequence informationMol. Phylogenet. Evol.15269282CrossRefPubMedGoogle Scholar
  57. Poran, N. S., Coss, R. G. 1990Development of antisnake defenses in California ground squirrels (Spermophilus beecheyi). I. Behavioral and immunological relationshipsBehaviour112222245Google Scholar
  58. Poran, N. S., Coss, R. G., Benjamini, E. 1987Resistance of California ground squirrels (Spermophilus beecheyi) to the venom of the Northern Pacific rattlesnake (Crotalus viridis oreganus): A study of adaptive variationToxicon25767777CrossRefPubMedGoogle Scholar
  59. Rowe, M. P., Owings, D. H. 1990Probing, assessment, and management during interactions between ground squirrels and rattlesnakes. 1. Risks related to rattlesnake size and body temperatureEthology86237249Google Scholar
  60. Rowe, M. P., Owings, D. H. 1996Probing, assessment, and management during interactions between ground squirrels (Rodentia: Sciuridae) and rattlesnakes (Squamata: Viperidae). 2. Cues afforded by rattlesnake rattlingEthology102856874Google Scholar
  61. Russel, F. E. 1980Snake Venom PoisoningLippincottPhiladelphiaGoogle Scholar
  62. Sasa, M. 1999Diet and snake venom evolution: Can local selection alone explain intraspecific venom variation?Toxicon37249252CrossRefPubMedGoogle Scholar
  63. Sasa, M. 1999ReplyToxicon37259260CrossRefGoogle Scholar
  64. Soto, J. G., Pérez, J. C., Minton, S. A. 1988Proteolytic, hemorrhagic and hemolytic activities of snake venomsToxicon26875882CrossRefPubMedGoogle Scholar
  65. Stock, C. 1918The Pleistocene fauna from Hawver caveUniv. Calif. Bull. Dept. Geol.10461515Google Scholar
  66. Swaisgood, R. R., Rowe, M. P, Owings, D. H. 1999Assessment of rattlesnake dangerousness by California ground squirrels: Exploitation of cues from rattling soundsAnim. Behav.5713011310CrossRefPubMedGoogle Scholar
  67. Thomas, R. G., Pough, F. H. 1979The effect of rattlesnake venom on digestion of preyToxicon17221228CrossRefPubMedGoogle Scholar
  68. Thompson, J. N. 1994The Coevolutionary ProcessUniversity of Chicago PressChicagoGoogle Scholar
  69. Thompson, J. N. 1997Evaluating the dynamics of coevolution among geographically structured populationsEcology7816191623Google Scholar
  70. Thwin, M. M., Gopalakrishnakone, P. 1998Snake envenomation and protective natural endogenous proteins: a mini review of the recent developments (1991–1997)Toxicon3614711482CrossRefPubMedGoogle Scholar
  71. Tomihara, Y., Yonaha, K., Nozaki, M., Yoshita, C. 1990Neutralization of hemorrhagic snake venoms by sera of Trimesurus flavoviridis (Habu), Herpestes edwardsii (mongoose) and Dinodon semicarinatus (Akamata)Toxicon28989991CrossRefPubMedGoogle Scholar
  72. Tu, A. T. 1991Tissue damaging effects by snake venoms: Hemorrhage and myonecrosisTu, A. T. eds. Handbook of Natural Toxins, Vol. 5, Reptile Venoms and ToxinsMarcel DekkerNew York297347Google Scholar
  73. Wüster, W., Daltry, J. C., Thorpe, R. S. 1999Can diet explain intraspecific venom variation? Reply to Sasa.Toxicon37253258CrossRefGoogle Scholar
  74. Zar, J. H. 1984Biostatistical AnalysisPrentice-HallEnglewood Cliffs, NJGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • James E. Biardi
    • 1
  • David C. Chien
    • 2
  • Richard G. Coss
    • 2
  1. 1.PROF Postdoctoral Program and Department of ChemistryUniversity of CaliforniaDavisUSA
  2. 2.Department of PsychologyUniversity of CaliforniaDavisUSA

Personalised recommendations