Seasonal Variations in Femoral Gland Secretions Reveals some Unexpected Correlations Between Protein and Lipid Components in a Lacertid Lizard

  • Marco MangiacottiEmail author
  • Stefano Pezzi
  • Marco Fumagalli
  • Alan Jioele Coladonato
  • Patrizia d’Ettorre
  • Chloé Leroy
  • Xavier Bonnet
  • Marco A. L. Zuffi
  • Stefano Scali
  • Roberto Sacchi


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.


Chemical communication Season Testosterone Quality Identity Femoral glands Cosinor models Lizards Podarcis muralis 



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.

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  1. Aguilar PM, Labra A, Niemeyer HM (2009) Chemical self-recognition in the lizard Liolaemus fitzgeraldi. J Ethol 27:181–184. CrossRefGoogle Scholar
  2. Aitchison J (1982) The statistical analysis of compositional data. J R Stat Soc Ser B 44:139–177Google Scholar
  3. Alberts AC (1990) Chemical properties of femoral gland secretions in the desert iguana, Dipsosaurus dorsalis. J Chem Ecol 16:13–25. CrossRefGoogle Scholar
  4. Alberts AC (1993) Chemical and behavioral studies of femoral gland secretions in iguanid lizards. Brain Behav Evol 41:255–260. CrossRefGoogle Scholar
  5. Alberts AC, Werner DI (1993) Chemical recognition of unfamiliar conspecifics by green iguanas: functional significance of different signal components. Anim Behav 46:197–199. CrossRefGoogle Scholar
  6. 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. CrossRefGoogle Scholar
  7. 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. CrossRefGoogle Scholar
  8. Alberts AC, Phillips JA, Werner DI (1993) Sources of intraspecific variability in the protein composition of lizard femoral gland secretions. Copeia 1993:775–781CrossRefGoogle Scholar
  9. Baeckens S (2019) Evolution of animal chemical communication : insights from non-model species and phylogenetic comparative methods. Belgian J Zool 149:63–93. CrossRefGoogle Scholar
  10. 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. CrossRefGoogle Scholar
  11. 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. CrossRefGoogle Scholar
  12. 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. CrossRefGoogle Scholar
  13. Bonnet X, Naulleau G (1996) Are body reserves important for reproduction in male dark Green snakes (Colubridae: Coluber viridiflavus)? Herpetologica 52:137–146Google Scholar
  14. 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. CrossRefGoogle Scholar
  15. 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 33–54Google Scholar
  16. 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. CrossRefGoogle Scholar
  17. Chamut S, Jahn GA, Arce OEA, Manes ME (2012) Testosterone and reproductive activity in the male tegu lizard, Tupinambis merianae. Herpetol Conserv Biol 7:299–305Google Scholar
  18. 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. CrossRefGoogle Scholar
  19. Cornelissen G (2014) Cosinor-based rhythmometry. Theor Biol Med Model 11:16CrossRefGoogle Scholar
  20. Crews D (1984) Gamete production, sex hormone secretion, and mating behavior uncoupled. Horm Behav 18:22–28. CrossRefGoogle Scholar
  21. 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. CrossRefGoogle Scholar
  22. 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. CrossRefGoogle Scholar
  23. 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. CrossRefGoogle Scholar
  24. 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. CrossRefGoogle Scholar
  25. Gan F, Ruan G, Mo J (2006) Baseline correction by improved iterative polynomial fitting with automatic threshold. Chemom Intell Lab Syst 82:59–65. CrossRefGoogle Scholar
  26. 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 497–513Google Scholar
  27. Grafen A (1990) Biological signals as handicaps. J Theor Biol 144:517–546. CrossRefGoogle Scholar
  28. 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. CrossRefGoogle Scholar
  29. 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. CrossRefGoogle Scholar
  30. 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. CrossRefGoogle Scholar
  31. 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 155–178Google Scholar
  32. Kéry M (2010) Introduction to winBUGS for ecologists: a Bayesian approach to regression, ANOVA, mixed models and related analyses, 1st edn. Elsevier Inc., AmsterdamGoogle Scholar
  33. Kruschke JK (2010) Bayesian data analysis. Wiley Interdiscip Rev Cogn Sci 1:658–676. CrossRefGoogle Scholar
  34. 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. CrossRefGoogle Scholar
  35. 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. CrossRefGoogle Scholar
  36. 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. CrossRefGoogle Scholar
  37. López P, Gabirot M, Martín J (2009) Immune activation affects chemical sexual ornaments of male Iberian wall lizards. Naturwissenschaften 96:65–69. CrossRefGoogle Scholar
  38. 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. Google Scholar
  39. Magnhagen C (1991) Predation risk as a cost of reproduction. Trends Ecol Evol 6:183–186. CrossRefGoogle Scholar
  40. 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. Google Scholar
  41. 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. CrossRefGoogle Scholar
  42. 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. CrossRefGoogle Scholar
  43. 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. CrossRefGoogle Scholar
  44. Martín J, López P (2007) Scent may signal fighting ability in male Iberian rock lizards. Biol Lett 3:125–127. CrossRefGoogle Scholar
  45. Martín J, López P (2015) Condition-dependent chemosignals in reproductive behavior of lizards. Horm Behav 68:14–24. CrossRefGoogle Scholar
  46. 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. CrossRefGoogle Scholar
  47. Martín J, Moreira PL, López P (2007b) Status-signalling chemical badges in male Iberian rock lizards. Funct Ecol 21:568–576. CrossRefGoogle Scholar
  48. 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. CrossRefGoogle Scholar
  49. 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. CrossRefGoogle Scholar
  50. 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. CrossRefGoogle Scholar
  51. 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. CrossRefGoogle Scholar
  52. 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. CrossRefGoogle Scholar
  53. McLean GS, Lee AK, Wilson KJ (1973) A simple method of obtaining blood from lizards. Copeia 1973:338–339CrossRefGoogle Scholar
  54. Meredith M, Kruschke J (2018) HDInterval: highest (posterior) density intervals. R package version 0.2.0. Accessed 27 Aug 2018
  55. 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. CrossRefGoogle Scholar
  56. 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. CrossRefGoogle Scholar
  57. 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. CrossRefGoogle Scholar
  58. Plummer M (2008) Penalized loss functions for Bayesian model comparison. Biostatistics 9:523–539. CrossRefGoogle Scholar
  59. Poncet P (2012) modeest: Mode Estimation. R package version 2.3.2. Accessed 5 March 2019
  60. Poynton C (2012) Digital video and HD: algorithms and interfaces. Elsevier Science, AmsterdamGoogle Scholar
  61. R Core Team (2018) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. Accessed 9 May 2018
  62. Randall D, Burggren W, French K (1997) Eckert animal physiology: mechanisms and adaptations, 5th edn. New York W.H. Freeman and Co, New YorkGoogle Scholar
  63. Ranke J (2018) chemCal: calibration functions for analytical chemistry. R package version 0.2.1. Accessed 8 Aug 2018
  64. Refinetti R, Cornélissen G, Halberg F (2007) Procedures for numerical analysis of circadian rhythms. Biol Rhythm Res 38:275–325. CrossRefGoogle Scholar
  65. 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. Google Scholar
  66. 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. CrossRefGoogle Scholar
  67. 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. CrossRefGoogle Scholar
  68. 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. CrossRefGoogle Scholar
  69. 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. CrossRefGoogle Scholar
  70. 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. CrossRefGoogle Scholar
  71. Schwenk K (1995) Of tongues and noses: chemoreception in lizards and snakes. Trends Ecol Evol 10:7–12. CrossRefGoogle Scholar
  72. 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. CrossRefGoogle Scholar
  73. 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. CrossRefGoogle Scholar
  74. Su U, Yajima M (2015) R2jags: Using R to run “JAGS”. R package version 0.5–7. Accessed 9 May 2018
  75. Tibbetts EA, Dale J (2007) Individual recognition: it is good to be different. Trends Ecol Evol 22:529–537. CrossRefGoogle Scholar
  76. 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. CrossRefGoogle Scholar
  77. Wenig P, Odermatt J (2010) OpenChrom: a cross-platform open source software for the mass spectrometric analysis of chromatographic data. BMC Bioinformatics 11:405. CrossRefGoogle Scholar
  78. West-Eberhard MJ (1979) Sexual selection, social competition, and evolution. Proc Am Philos Soc 123:222–234Google Scholar
  79. 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–1061073CrossRefGoogle Scholar
  80. Whiting MJ, While GM (2017) Sociality in lizards. In: Rubenstein DR, Abbot P (eds) Comparative Social Evolution. Cambridge University Press, Cambridge, pp 354–389Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Marco Mangiacotti
    • 1
    • 2
    Email author
  • Stefano Pezzi
    • 1
  • Marco Fumagalli
    • 3
  • Alan Jioele Coladonato
    • 1
  • Patrizia d’Ettorre
    • 4
  • Chloé Leroy
    • 4
  • Xavier Bonnet
    • 5
  • Marco A. L. Zuffi
    • 6
  • Stefano Scali
    • 2
  • Roberto Sacchi
    • 1
  1. 1.Department of Earth and Environmental SciencesUniversity of PaviaPaviaItaly
  2. 2.Museo di Storia Naturale di MilanoMilanItaly
  3. 3.Department of Biology and Biotechnologies “L. Spallanzani”, Unit of BiochemistryUniversity of PaviaPaviaItaly
  4. 4.LEEC Laboratoire d’Ethologie Expérimentale et ComparéeVilletaneuseFrance
  5. 5.Centre d’Etudes Biologiques de ChizéCNRS UMR 7372 - Université de La RochelleVilliers-en-BoisFrance
  6. 6.Museo di Storia Naturale dell’Università di PisaCalciItaly

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