Abstract
Animals modulate intraspecific signal shape and intensity, notably during reproductive periods. Signal variability typically follows a seasonal scheme, traceable through the expression of visual, acoustic, chemical and behavioral patterns. The chemical channel is particularly important in lizards, as demonstrated by well-developed epidermal glands in the cloacal region that secrete lipids and proteins recognized by conspecifics. In males, the seasonal pattern of gland activity is underpinned by variation of circulating androgens. Changes in the composition of lipid secretions convey information about the signaler’s quality (e.g., size, immunity). Presumably, individual identity is associated with a protein signature present in the femoral secretions, but this has been poorly investigated. For the first time, we assessed the seasonal variability of the protein signal in relation to plasma testosterone level (T), glandular activity and the concentration of provitamin D3 in the lipid fraction. We sampled 174 male common wall lizards (Podarcis muralis) over the entire activity season. An elevation of T was observed one to two months before the secretion peak of lipids during the mating season; such expected delay between hormonal fluctuation and maximal physiological response fits well with the assumption that provitamin D3 indicates individual quality. One-dimensional electrophoretic analysis of proteins showed that gel bands were preserved over the season with an invariant region; a result in agreement with the hypothesis that proteins are stable identity signals. However, the relative intensity of bands varied markedly, synchronously with that of lipid secretion pattern. These variations of protein secretion suggest additional roles of proteins, an issue that requires further studies.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aguilar PM, Labra A, Niemeyer HM (2009) Chemical self-recognition in the lizard Liolaemus fitzgeraldi. J Ethol 27:181–184. https://doi.org/10.1007/s10164-008-0088-x
Aitchison J (1982) The statistical analysis of compositional data. J R Stat Soc Ser B 44:139–177
Alberts AC (1990) Chemical properties of femoral gland secretions in the desert iguana, Dipsosaurus dorsalis. J Chem Ecol 16:13–25. https://doi.org/10.1007/BF01021264
Alberts AC (1993) Chemical and behavioral studies of femoral gland secretions in iguanid lizards. Brain Behav Evol 41:255–260. https://doi.org/10.1159/000113847
Alberts AC, Werner DI (1993) Chemical recognition of unfamiliar conspecifics by green iguanas: functional significance of different signal components. Anim Behav 46:197–199. https://doi.org/10.1006/anbe.1993.1177
Alberts AC, Pratt NC, Phillips JA (1992a) Seasonal productivity of lizard femoral glands: relationship to social dominance and androgen levels. Physiol Behav 51:729–733. https://doi.org/10.1016/0031-9384(92)90109-F
Alberts AC, Sharp TR, Werner DI, Weldon PJ (1992b) Seasonal variation of lipids in femoral gland secretions of male green iguanas (Iguana iguana). J Chem Ecol 18:703–712. https://doi.org/10.1007/BF00994608
Alberts AC, Phillips JA, Werner DI (1993) Sources of intraspecific variability in the protein composition of lizard femoral gland secretions. Copeia 1993:775–781
Baeckens S (2019) Evolution of animal chemical communication : insights from non-model species and phylogenetic comparative methods. Belgian J Zool 149:63–93. https://doi.org/10.26496/bjz.2019.31
Baeckens S, Huyghe K, Palme R, Van Damme R (2017) Chemical communication in the lacertid lizard Podarcis muralis: the functional significance of testosterone. Acta Zool 98:94–103. https://doi.org/10.1111/azo.12160
Baeckens S, Martín J, García-Roa R, Van Damme R (2018) Sexual selection and the chemical signal design of lacertid lizards. Zool J Linnean Soc 183:445–457. https://doi.org/10.1093/zoolinnean/zlx075
Bickel DR, Frühwirth R (2006) On a fast, robust estimator of the mode: comparisons to other robust estimators with applications. Comput Stat Data Anal 50:3500–3530. https://doi.org/10.1016/j.csda.2005.07.011
Bonnet X, Naulleau G (1996) Are body reserves important for reproduction in male dark Green snakes (Colubridae: Coluber viridiflavus)? Herpetologica 52:137–146
Carazo P, Font E, Desfilis E (2008) Beyond “nasty neighbours” and “dear enemies”? Individual recognition by scent marks in a lizard (Podarcis hispanica). Anim Behav 76:1953–1963. https://doi.org/10.1016/j.anbehav.2008.08.018
Carretero MA (2006) Reproductive cycles in Mediterranean lacertids: plasticity and constraints. In: Corti C, Lo Cascio P, Biaggini M (eds) Mainland and insular lacertid lizards: a mediterannean perspective. Firenze University Press, Firenze, pp 33–54
Carretero MA, Ribeiro R, Barbosa D, Sá-Sousa P, Harris DJ (2006) Spermatogenesis in two iberian podarcis lizards: relationships with male traits. Anim Biol 56:1–12. https://doi.org/10.1163/157075606775904759
Chamut S, Jahn GA, Arce OEA, Manes ME (2012) Testosterone and reproductive activity in the male tegu lizard, Tupinambis merianae. Herpetol Conserv Biol 7:299–305
Cooper WE (1991) Responses to prey chemicals by a lacertid lizard, Podarcis muralis: prey chemical discrimination and poststrike elevation in tongue-flick rate. J Chem Ecol 17:849–863. https://doi.org/10.1007/BF01395595
Cornelissen G (2014) Cosinor-based rhythmometry. Theor Biol Med Model 11:16
Crews D (1984) Gamete production, sex hormone secretion, and mating behavior uncoupled. Horm Behav 18:22–28. https://doi.org/10.1016/0018-506X(84)90047-3
Dale J, Lank DB, Reeve HK (2001) Signaling individual identity versus quality: a model and case studies with ruffs, Queleas, and house finches. Am Nat 158:75–86. https://doi.org/10.1086/320861
Dawley EM, Crowder J (1995) Sexual and seasonal differences in the vomeronasal epithelium of the red-backed salamander (Plethodon cinereus). J Comp Neurol 359:382–390. https://doi.org/10.1002/cne.903590303
Font E, Barbosa D, Sampedro C, Carazo P (2012) Social behavior, chemical communication, and adult neurogenesis: studies of scent mark function in Podarcis wall lizards. Gen Comp Endocrinol 177:9–17. https://doi.org/10.1016/j.ygcen.2012.02.015
Gabirot M, López P, Martín J, de Fraipont M, Heulin B, Sinervo B, Clobert J (2008) Chemical composition of femoral secretions of oviparous and viviparous types of male common lizards Lacerta vivipara. Biochem Syst Ecol 36:539–544. https://doi.org/10.1016/j.bse.2008.03.006
Gan F, Ruan G, Mo J (2006) Baseline correction by improved iterative polynomial fitting with automatic threshold. Chemom Intell Lab Syst 82:59–65. https://doi.org/10.1016/j.chemolab.2005.08.009
Garfin DE (2009) One-dimensional gel electrophoresis. In: Burgess RR, Deutscher MPBT-M in E (eds) Methods in Enzymology, 2nd edn. Academic Press, Cambridge pp 497–513
Grafen A (1990) Biological signals as handicaps. J Theor Biol 144:517–546. https://doi.org/10.3102/0002831211408061
Graham SP, Earley RL, Hoss SK, Schuett GW, Grober MS (2008) The reproductive biology of male cottonmouths (Agkistrodon piscivorus): do plasma steroid hormones predict the mating season? Gen Comp Endocrinol 159:226–235. https://doi.org/10.1016/j.ygcen.2008.09.002
Gribbins KM, Gist DH (2003) Cytological evaluation of spermatogenesis within the germinal epithelium of the male European Wall lizard, Podarcis muralis. J Morphol 258:296–306. https://doi.org/10.1002/jmor.10143
Heathcote RJP, Bell E, d’Ettorre P, While GM, Uller T (2014) The scent of sun worship: basking experience alters scent mark composition in male lizards. Behav Ecol Sociobiol 68:861–870. https://doi.org/10.1007/s00265-014-1700-4
Johnstone RA (1997) The evolution of animal signals. In: Krebs JR, Davies NB (eds) Behavioural ecology: an evolutionary approach, fourth. Blackwell Publishing Ltd, Malden, pp 155–178
Kéry M (2010) Introduction to winBUGS for ecologists: a Bayesian approach to regression, ANOVA, mixed models and related analyses, 1st edn. Elsevier Inc., Amsterdam
Kruschke JK (2010) Bayesian data analysis. Wiley Interdiscip Rev Cogn Sci 1:658–676. https://doi.org/10.1002/wcs.72
López P, Martín J (2001) Fighting roles and rival recognition reduce costs of aggression in male lizards, Podarcis hispanica. Behav Ecol Sociobiol 49:111–116. https://doi.org/10.1007/s002650000288
López P, Martín J (2005) Female Iberian wall lizards prefer male scents that signal a better cell-mediated immune response. Biol Lett 1:404–406. https://doi.org/10.1098/rsbl.2005.0360
López P, Amo L, Martín J (2006) Reliable signaling by chemical cues of male traits and health state in male lizards, Lacerta monticola. J Chem Ecol 32:473–488. https://doi.org/10.1007/s10886-005-9012-9
López P, Gabirot M, Martín J (2009) Immune activation affects chemical sexual ornaments of male Iberian wall lizards. Naturwissenschaften 96:65–69. https://doi.org/10.1007/s00114-008-0451-3
MacGregor HEA, Lewandowsky RAM, D’Ettorre P, Davies NW, Leroy C, Uller T, While GM (2017) Chemical communication, sexual selection, and introgression in wall lizards. Evolution (N Y) 71:2327–2343. https://doi.org/10.1111/evo.13317
Magnhagen C (1991) Predation risk as a cost of reproduction. Trends Ecol Evol 6:183–186. https://doi.org/10.1016/0169-5347(91)90210-O
Mangiacotti M, Fumagalli M, Scali S, Zuffi MAL, Cagnone M, Salvini R, Sacchi R (2017) Inter- and intra-population variability of the protein content of femoral gland secretions from a lacertid lizard. Curr Zool 63:657–665. https://doi.org/10.1093/cz/zow113
Mangiacotti M, Fumagalli M, Cagnone M, Viglio S, Bardoni AM, Scali S, Sacchi R (2019a) Morph-specific protein patterns in the femoral gland secretions of a colour polymorphic lizard. Sci Rep 9:8412. https://doi.org/10.1038/s41598-019-44889-7
Mangiacotti M, Gaggiani S, Coladonato AJ, Scali S, Zuffi MAL, Sacchi R (2019b) First experimental evidence that proteins from femoral glands convey identity related information in a lizard. Acta Ethol 22:57–65. https://doi.org/10.1007/s10211-018-00307-1
Martín J, López P (2006) Links between male quality, male chemical signals, and female mate choice in Iberian rock lizards. Funct Ecol 20:1087–1096. https://doi.org/10.1111/j.1365-2435.2006.01183.x
Martín J, López P (2007) Scent may signal fighting ability in male Iberian rock lizards. Biol Lett 3:125–127. https://doi.org/10.1098/rsbl.2006.0589
Martín J, López P (2015) Condition-dependent chemosignals in reproductive behavior of lizards. Horm Behav 68:14–24. https://doi.org/10.1016/j.yhbeh.2014.06.009
Martín J, López P, Gabirot M, Pilz KM (2007a) Effects of testosterone supplementation on chemical signals of male Iberian wall lizards: consequences for female mate choice. Behav Ecol Sociobiol 61:1275–1282. https://doi.org/10.1007/s00265-007-0358-6
Martín J, Moreira PL, López P (2007b) Status-signalling chemical badges in male Iberian rock lizards. Funct Ecol 21:568–576. https://doi.org/10.1111/j.1365-2435.2007.01262.x
Martín J, Amo L, López P (2008) Parasites and health affect multiple sexual signals in male common wall lizards, Podarcis muralis. Naturwissenschaften 95:293–300. https://doi.org/10.1007/s00114-007-0328-x
Martín J, Castilla AM, López P, Al-Jaidah M, Al-Mohannadi SF, Al-Hemaidi AAM (2016) Chemical signals in desert lizards: are femoral gland secretions of male and female spiny-tailed lizards, Uromastyx aegyptia microlepis adapted to arid conditions? J Arid Environ 127:192–198. https://doi.org/10.1016/j.jaridenv.2015.12.004
Martins EP, Ord TJ, Slaven J, Wright JL, Housworth EA (2006) Individual, sexual, seasonal, and temporal variation in the amount of sagebrush lizard scent marks. J Chem Ecol 32:881–893. https://doi.org/10.1007/s10886-006-9029-8
Mayerl C, Baeckens S, Van Damme R (2015) Evolution and role of the follicular epidermal gland system in non-ophidian squamates. Amphibia-Reptilia 36:185–206. https://doi.org/10.1163/15685381-00002995
McGraw KJ, Hill GE (2004) Plumage color as a dynamic trait: carotenoid pigmentation of male house finches ( Carpodacus mexicanus ) fades during the breeding season. Can J Zool 82:734–738. https://doi.org/10.1139/z04-043
McLean GS, Lee AK, Wilson KJ (1973) A simple method of obtaining blood from lizards. Copeia 1973:338–339
Meredith M, Kruschke J (2018) HDInterval: highest (posterior) density intervals. R package version 0.2.0. https://CRAN.R-project.org/package=HDInterval. Accessed 27 Aug 2018
Paul MJ, Zucker I, Schwartz WJ (2008) Tracking the seasons: the internal calendars of vertebrates. Philos Trans R Soc B Biol Sci 363:341–361. https://doi.org/10.1098/rstb.2007.2143
Peig J, Green AJ (2009) New perspectives for estimating body condition from mass/length data: the scaled mass index as an alternative method. Oikos 118:1883–1891. https://doi.org/10.1111/j.1600-0706.2009.17643.x
Pellitteri-Rosa D, Martín J, López P, Bellati A, Sacchi R, Fasola M, Galeotti P (2014) Chemical polymorphism in male femoral gland secretions matches polymorphic coloration in common wall lizards (Podarcis muralis). Chemoecology 24:67–78. https://doi.org/10.1007/s00049-014-0148-3
Plummer M (2008) Penalized loss functions for Bayesian model comparison. Biostatistics 9:523–539. https://doi.org/10.1093/biostatistics/kxm049
Poncet P (2012) modeest: Mode Estimation. R package version 2.3.2. https://CRAN.R-project.org/package=modeest. Accessed 5 March 2019
Poynton C (2012) Digital video and HD: algorithms and interfaces. Elsevier Science, Amsterdam
R Core Team (2018) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. https://www.R-project.org/. Accessed 9 May 2018
Randall D, Burggren W, French K (1997) Eckert animal physiology: mechanisms and adaptations, 5th edn. New York W.H. Freeman and Co, New York
Ranke J (2018) chemCal: calibration functions for analytical chemistry. R package version 0.2.1. https://CRAN.R-project.org/package=chemCal. Accessed 8 Aug 2018
Refinetti R, Cornélissen G, Halberg F (2007) Procedures for numerical analysis of circadian rhythms. Biol Rhythm Res 38:275–325. https://doi.org/10.1080/09291010600903692
Roig JM, Llorente GA, Carretero MA (2000) Reproductive cycle in a Pyrenean oviparous population of the common lizard (Zootoca vivipara). Netherlands J Zool 50:15–27. https://doi.org/10.1163/156854200x00243
Sacchi R, Pupin F, Gentilli A, Rubolini D, Scali S, Fasola M, Galeotti P (2009) Male-male combats in a polymorphic lizard: residency and size, but not color, affect fighting rules and contest outcome. Aggress Behav 35:274–283. https://doi.org/10.1002/ab.20305
Sacchi R, Scali S, Pellitteri-Rosa D, Pupin F, Gentilli A, Tettamanti S, Cavigioli L, Racina L, Maiocchi V, Galeotti P, Fasola M (2010) Photographic identification in reptiles: a matter of scales. Amphibia-Reptilia 31:489–502. https://doi.org/10.1163/017353710X521546
Sacchi R, Pellitteri-Rosa D, Capelli A, Ghitti M, Di Paoli A, Bellati A, Scali S, Galeotti P, Fasola M (2012) Studying the reproductive biology of the common wall lizard using ultrasonography. J Zool 287:301–310. https://doi.org/10.1111/j.1469-7998.2012.00917.x
Sacchi R, Ghitti M, Scali S, Mangiacotti M, Zuffi MAL, Sannolo M, Coladonato AJ, Pasquesi G, Bovo M, Pellitteri-Rosa D (2015) Common Wall lizard females (Podarcis muralis) do not actively choose males based on their colour morph. Ethology 121:1145–1153. https://doi.org/10.1111/eth.12431
Sacchi R, Scali S, Mangiacotti M, Sannolo M, Zuffi MAL, Pupin F, Gentilli A, Bonnet X (2017) Seasonal variations of plasma testosterone among colour-morph common wall lizards (Podarcis muralis). Gen Comp Endocrinol 240:114–120. https://doi.org/10.1016/j.ygcen.2016.09.012
Schwenk K (1995) Of tongues and noses: chemoreception in lizards and snakes. Trends Ecol Evol 10:7–12. https://doi.org/10.1016/S0169-5347(00)88953-3
Sillero N, Campos J, Bonardi A, Corti C, Creemers R, Crochet PA, Isailović JC, Denoël M, Ficetola GF, Gonçalves J, Kuzmin S, Lymberakis P, De Pous P, Rodríguez A, Sindaco R, Speybroeck J, Toxopeus B, Vieites DR, Vences M (2014) Updated distribution and biogeography of amphibians and reptiles of Europe. Amphibia-Reptilia 35:1–31. https://doi.org/10.1163/15685381-00002935
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid [published erratum appears in Anal Biochem 1987 May 15;163(1):279]. Anal Biochem 150:76–85. https://doi.org/10.1016/0003-2697(85)90442-7
Su U, Yajima M (2015) R2jags: Using R to run “JAGS”. R package version 0.5–7. https://CRAN.R-project.org/package=R2jags. Accessed 9 May 2018
Tibbetts EA, Dale J (2007) Individual recognition: it is good to be different. Trends Ecol Evol 22:529–537. https://doi.org/10.1016/j.tree.2007.09.001
van Wyk JH (1990) Seasonal testicular activity and morphometric variation in the femoral glands of the lizard Cordylus polyzonus polyzonus (Sauria: Cordylidae). J Herpetol 24:405–409. https://doi.org/10.2307/1565059
Wenig P, Odermatt J (2010) OpenChrom: a cross-platform open source software for the mass spectrometric analysis of chromatographic data. BMC Bioinformatics 11:405. https://doi.org/10.1186/1471-2105-11-405
West-Eberhard MJ (1979) Sexual selection, social competition, and evolution. Proc Am Philos Soc 123:222–234
Westneat DF, Birkhead TR (1998) Alternative hypotheses linking the immune system and mate choice for good genes. Proc R Soc Lond Ser B Biol Sci 265:1065 LP–1061073
Whiting MJ, While GM (2017) Sociality in lizards. In: Rubenstein DR, Abbot P (eds) Comparative Social Evolution. Cambridge University Press, Cambridge, pp 354–389
Acknowledgments
The study was performed in accordance with the European and Italian laws on animal use in scientific research, and all the protocols have been authorized by Italian Environmental Ministry (Aut. Prot. PNM-2015-0010423, PNM-2016-0002154). This research was funded by the FRG_2016 (Italian Ministry of Education, University and Research - MIUR) to Roberto Sacchi. We would like to thank Matteo Panaccio for his help during fieldwork, Lorenzo Balestrazzi for his help in lab analysis, the Associate Editor and two anonymous reviewers for their constructive comments on the early version of the manuscript. The authors declared no competing interests.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
ESM 1
(PDF 110 kb)
Rights and permissions
About this article
Cite this article
Mangiacotti, M., Pezzi, S., Fumagalli, M. et al. Seasonal Variations in Femoral Gland Secretions Reveals some Unexpected Correlations Between Protein and Lipid Components in a Lacertid Lizard. J Chem Ecol 45, 673–683 (2019). https://doi.org/10.1007/s10886-019-01092-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10886-019-01092-2