Variability of testes morphology and the presence of testis-ova in the European blind cave salamander (Proteus anguinus)

Authors

  • Lilijana Bizjak Mali

DOI:

https://doi.org/10.14720/abs.60.1.15669

Keywords:

testis, morphology, spermatogenesis, proteus, Proteus anguinus

Abstract

The European blind cave salamander, Proteus anguinus, is a neotenic, permanently aquatic amphibian with a very long life span, late sexual maturity, and the ability to reproduce for at least 30 years. Proteus is considered to be vulnerable species, and yet very little is known about its reproductive biology. The objective of this study is to describe the detailed morphology of the testes of adult Proteus and determine the maturation state of the gonads and gametogenesis with respect to body size and seasonality. This research showed that testis size increases with adult male body length, but the shape and meiotic condition of the testes are highly variable and independent of the length of the specimen. The testis of Proteus has a simple cystic type of organization in which cysts are enclosed in lobules, with synchronous maturation of the germ cells within each cyst. Spermatogenesis progresses in a caudocephalic direction within the testis, as in other salamanders, and appears to be seasonal, despite the fact that Proteus is a cave animal living in stable environmental conditions. Surprisingly, the testes of approximately one third of the specimens, regardless of
their morphology or meiotic condition, contained testis-ova located randomly among groups of spermatogonia and spermatocytes. These testes-ova contained germinal vesicles with lampbrush chromosomes, and thus correspond to primary oocytes. The
presence of testis-ova may be related to a sex-chromosome turnover involving X-Y translocation that was recently discovered in Proteus.

References

Bizjak Mali, L., Sepčić, K., Bulog, B., 2013. Long-term starvation in cave salamander effects on liver ultrastructure and energy reserve mobilization. J. Morph., 274(8), 887-900. DOI: https://doi.org/10.1002/jmor.20145

Bizjak Mali, L., Bulog, B., 2010. Ultrastructure of previtellogene oocytes in the neotenic cave salamander Proteus anguinus anguinus (Amphibia, Urodela, Proteidae). Protoplasma, 246, 33-39. DOI: https://doi.org/10.1007/s00709-010-0117-9

Bizjak Mali, L., Talaber, I, Žibert, U., Bulog, B., 2010. Oogenesis in Proteus: stages of oocyte development. In: Moškrič, A. (ed.), Trontelj, P. (eds.). Abstract book. Postojna: Organizing committee, 20th International Conference on Subterranean Biology, pp. 116-117.

Bizjak Mali, L., Bulog, B., 2011. Follicular ovarian atresia in the olm (Proteus anguinus anguinus). In: MCM 2011 : [proceedings], 10th Multinational Congress on Microscopy 2011, Urbino, Italy. [S. l.]: Società Italiana Scienze Micriscopiche, pp. 295-296.

Bizjak Mali, L., Talaber, I, Žibert, U., Ceket, D., Habič, L., Bulog, B., 2013. Oogenesis of the olm. The Anatomical Record, 296, spec. feat. 1.

Briegleb, W. 1961. Die Spermatophore des Grottenolms. Zool Anz, 166, 87-91.

Briegleb, W. 1962. Zur Biologie und Ökologie des Grottenolmes (Proteus anguinus Laur. 1768). Z. Morph. Ökol., 51, 271-334. DOI: https://doi.org/10.1007/BF00407657

Bruce, R.C., 2003. Life history. In: Reproductive biology and phylogeny of Urodela. Sever D.M. (ed.). 1st edition, USA, Science Publichers, Inc., 477-525.

Bulog, B., Bizjak Mali, L., Kos, M., Mihajl, K., Prelovšek, P.M., Aljančič, G., 2000. Biology and functional morphology of Proteus anguinus (Amphibia, Caudata), Acta boil. Slov., 43(3), 85-102.

Bulog, B., Bizjak Mali, L., 2014. Olm - cave salamander. In: ŠTANGELJ, Mojmir (ed.), et al. Natural heritage of Bela krajina, Slovenia. Bela krajina Museum, pp. 176-187.

Callard, G.V., 1992. Autocrine and paracrine role of steroids during spermatogenesis: Studies in Squalus acanthias and Necturus maculosus. J. Experimental Zoo., 261, 132-142. DOI: https://doi.org/10.1002/jez.1402610204

Chan, L.M., 2003. Seasonality, microhabitat and cryptic variation in tropical salamander reproductive cycles. Biological Journal of the Linnean Society, 78, 489–496. DOI: https://doi.org/10.1046/j.0024-4066.2002.00157.x

De Sa, R., Berois, N., 1986. Spermatogenesis and Histology of the Testes of the Caecilian, Chthonerpeton indistinctum. Journal of Herpetology, 20 (4), 510-514. DOI: https://doi.org/10.2307/1564247

Dimitri, P., Pisano, C., 1989. Position effect variegation in Drosophila melanogaster: relationship between suppression effect and the amount of y chromosome. Genetics, 122, 793-800. DOI: https://doi.org/10.1093/genetics/122.4.793

Durand, J., Delay, B., 1981. Influence of temperature on the development of Proteus anguinus (Caudata: Proteidae) and relation with its habitat in the subterranean world. J. Thermal. Biol., 6 (1), 53–57. DOI: https://doi.org/10.1016/0306-4565(81)90044-9

Exbrayat, J.M., 2009. Oogenesis and female reproductive system in Amphibia – Gymnophiona. In: Reproduction of Amphibians. Ogielska M. (ed.). Poland, Zoological Institute University of WrocŁaw, Science publishers, pp. 305-342.

Ferguson-Smith, M., 1966. X-Y chromosomal interchange in the aetiology of true hermaphroditism and of XX Klinefelter’s syndrome. Lancet, 2, 475–476. DOI: https://doi.org/10.1016/S0140-6736(66)92778-4

Flament, S., Dumond, H., Chardard, D., Chesnel, A., 2009. Lifelong testicular differentiation in Pleurodeles waltl (Amphibia, Caudata). Reproductive biology and endocrinology, 7, 21. DOI: https://doi.org/10.1186/1477-7827-7-21

Gregory, T.R., 2001. The bigger the C-value, the larger the cell: genome size and red blood cell size in vertebrates. Blood Cells Mol. Dis., 27(5), 830-843. DOI: https://doi.org/10.1006/bcmd.2001.0457

Gregory, T.R., 2005. Animal Genome Size Database. http://www.genomesize.com

Griffing, A.H., Bowerman, J., Sessions, S.K., 2017. Histology reveals testicular oocytes and trematode cysts in the threatened Oregon spotted frog (Rana pretiosa). Northwestern Naturalist, 98, 24-32. DOI: https://doi.org/10.1898/NWN16-13.1

Gorički, Š., Trontelj, P., 2006. Structure and evolution of the mitochondrial control region and flanking sequences in the European cave salamander Proteus anguinus. Gene, 378, 31–41. DOI: https://doi.org/10.1016/j.gene.2006.04.016

Gorički, Š., Stanković, D., Snoj, A., Kuntner, M., Jeffery, W. J., Trontelj, P., Pavićević, M., Grizelj, Z., Năpăruş-Aljančič, M., Aljančič, G., 2017. Environmental DNA in subterranean biology: rangeextension and taxonomic implications for Proteus. Scientific Reports, 7, 45054. DOI: https://doi.org/10.1038/srep45054

Hecker, M., Murphy, M. B., Coady, K. K., Vileneuve, D. L., Jones, P. D., Carr, J. A., Van der Kraak, G., 2006. Terminology of gonadal anomalies in fish and amphibians resulting from chemical expousures. Rev. Environ. Contam. Toxicol., 187, 103-131. DOI: https://doi.org/10.1007/0-387-32885-8_3

Hayes, T. B., Collins, A., Lee, M., Mendoza, M., Noriega, N., Stuart, A.A., Vonk, A., 2002. Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses. Proc. Natl. Acad. Sci. U S A, 99(8), 5476-80. DOI: https://doi.org/10.1073/pnas.082121499

Hervant, F., Mathieu, J., Durand, J. P., 2001. Behavioural, physiological and metabolic responses to long-term starvation and refeeding in a blind cave-dwelling (Proteus anguinus) and a surfacedwelling (Euproctus asper) salamander. J. Exp. Biol., 204, 269–281. DOI: https://doi.org/10.1242/jeb.204.2.269

Humason, G.L., 1979. Animal Tissue Technique. San Francisco: W.H. Freeman. Humphrey, R.R., 1922. The multiple testis in Urodeles. Biological Bulletin, 4, 45-67. DOI: https://doi.org/10.2307/1536690

Juberthie, C., Durand, J., Dupuy, M., 1996. La Rreproduction des Protées (Proteus anguinus): Bilan de 35 ans D‘elevage dans les grotteslaboratories de Moulis et D‘aulignac. Mémoires de Biospéologie, Tome XXIII, pp. 53–56.

Kezer, J., 1962. The chromosome number of the Europan cave salamander Proteus anguinus Laurenti. Biološki vestnik, 10, 45-48.

Kobayashi, M., Iwasawa, H., 1976. Development of the testis in the frog Rana nigromaculata, with special reference to germ cell maturation. Copeia, 3, 461-467. DOI: https://doi.org/10.2307/1443360

Kobayashi, T., Kumakura, M., Yoshie, S., Sugishima, T., Horie, Y., 2014. Dynamics of testis-ova in a wild population of Japanese pond frogs, Rana nigromaculata. J. Exp. Zool., 00A, 1–6. DOI: https://doi.org/10.1002/jez.1898

Kos, M., Bulog, B., Röhlich, A.S.P., 2001. Immunocytochemical demonstration of visual pigments in the degenerate retinal and pineal photoreceptors of the blind cave salamander (Proteus anguinus). Cell Tissue Res., 303, 15–25. DOI: https://doi.org/10.1007/s004410000298

Kosai, P., Jiraungkoorskul, W., Sachamahithinant, C., Jiraungkoorskul, K., 2011. Induction of testisova in nile tilapia (Oreochromis niloticus) exposed to 17S-estradiol. Natural Science, 3, 227-233. DOI: https://doi.org/10.4236/ns.2011.33029

Lambert, M. R., Gillera, G. S. J., Barberb, L. B, Fitzgerald, K. C., Skellya, D. K., 2015. Suburbanization, estrogen contamination, and sex ratio in wild amphibian populations. PNAS, 112(38), 11881-11886. DOI: https://doi.org/10.1073/pnas.1501065112

Kiernan, J.A., 1990. Histological and Histochemical methods: Theory and practise. 2nd edition, Oxford, Pergamon press.

Langecker, T.G., 2000. The effects of continuous darkness on cave ecology and caverniculous evolution. In: Culver D.C. et al (eds.) Ecosystems of the world: subterranean ecosystems, 1st edn. Elsevier, Amsterdam, pp. 135–157.

Macgregor, H.C., Walker, M.H., 1973. The arrangement of chromosomes in nucleus of sperm from plethodontid salamanders. Chromosoma, 40, 243-262. DOI: https://doi.org/10.1007/BF00326179

Noble, G.K., 1931. The Biology of Amphibia. Dover Publ., N.Y. Ogielska, M., Bartmanska, J., 1998. Development of testes and differentiation of germ cells in water frogs of the Rana esculenta - Complex (Amphibia, Anura). Amphibia-reptilia, 20, 251-263. DOI: https://doi.org/10.1163/156853899507059

Ogielska, M., Bartmanska, J., 2009. Spermatogenesis and male reproductive system in Amphibia – Anura. In: Reproduction of Amphibians. Ogielska M. (ed.). Poland, Zoological Institute University of WrocŁaw, Science publishers, pp. 66-68.

Pierantoni, R., Cobellis, G., Meccariello, R., Palmiero, C., Fienga, G., Minucci, S., Fasano, S., 2002. The amphibian testis as model to study germ cell progression during spermatogenesis. Comparative Biochemistry and Physiology Part B, 132, 131–139. DOI: https://doi.org/10.1016/S1096-4959(01)00543-7

Presnell, J.K., Schreibman, M.P., 1997. Humason’s animal tissue techniques, 5th edn. The Johns Hopkins University Press, Baltimore.

Pudney, J., 1995. Spermatogenesis in nonmommalian vertebrates. Microscopy Research and Technique, 32, 459-497. DOI: https://doi.org/10.1002/jemt.1070320602

Pudney, J., Callard, V.G., 1984. Organization of Interstitial Tissue in the Testis of the Salamander Necturus maculosus (Caudata: Proteidae). J. Morph., 181, 87-95. DOI: https://doi.org/10.1002/jmor.1051810108

Scheltinga, D.M., Jamieson, B.G.M., 2003. The mature spermatozoon. In: Reproductive biology and phylogeny of Urodela. Sever D.M. (ed.). 1st edition, USA, Science Publichers, Inc., 204-274.

Schlegel, P.A., Steinfartz, S., Bulog, B., 2009. Non-visual sensory physiology and magnetic orientation in the Blind Cave Salamander, Proteus anguinus (and some other cave-dwelling urodele species). Review and new results on light-sensitivity and non-visual orientation in subterranean urodeles (Amphibia). Animal Biology, 59, 351–384. DOI: https://doi.org/10.1163/157075609X454971

Sessions, K.S., 2008. Evolutionary cytogenetics in salamanders. Chromosome Research, 16, 183-201. DOI: https://doi.org/10.1007/s10577-007-1205-3

Sessions, K.S., Bizjak Mali, L., Green, D. M., Trifonov, V., Ferguson-Smith, M.A., 2016. Evidence for sex chromosome turnover in proteid salamanders. Cytogenetic and genome research, 148(4), 305-313. DOI: https://doi.org/10.1159/000446882

Sever, M., 1974. The occurrence of multiple testis in the genus Eurycea (Amphibia: Plethodontidae). Herpetologica, 30(2), 187-193.

Sever, M., 2002. Female sperm storage in amphibians. J Exp Zool, 292, 165–179. DOI: https://doi.org/10.1002/jez.1152

Shoop, R. C., 1965. Aspects of reproduction in Louisiana Necturus populations. Am. Midl. Nat., 74, 357–367. DOI: https://doi.org/10.2307/2423266

Sket, B., Arntzen, J.W., 1994. A black, non-troglomorphic amphibian from the karst of Slovenia: Proteus anguinus parkelj n. ssp. (Urodela: Proteidae). Bijdragen tot de Dierkunde, 64, 33-53. DOI: https://doi.org/10.1163/26660644-06401002

Sket, B., 1997. Distribution of Proteus (Amphibia: Urodela: Proteidae) and its possible explanation. J. Biogeogr., 24, 263–280. DOI: https://doi.org/10.1046/j.1365-2699.1997.00103.x

Smita, M, Oommen, O.V., Jancy, M.G., Akbarsha, M.A., 2004. Stages in spermatogenesis of two species of caecilians, Ichthyophis tricolor and Uraeotyphlus cf. narayani (Amphibia: Gymnophiona): Light and electron microscopic study. J. Morphol., 261, 92–104. DOI: https://doi.org/10.1002/jmor.10231

Uribe, M.C.A., 2003. The testes, spermatogenesis and male reproductive ducts. In: Reproductive biology and phylogeny of Urodela. Sever D.M. (ed.). 1st edition, USA, Science Publichers, Inc., 183-202.

Uribe, M.C.A., 2009. Spermatogenesis and male reproductive system in Amphibia – Urodela. In: Reproduction of Amphibians. Ogielska M. (ed.). Poland, Zoological Institute University of WrocŁaw, Science publishers, 100-124.

Uribe, M.C., Mejía-Roa, V., 2014. Testicular structure and germ cells morphology in salamanders. Spermatogenesis, 4(3), e988090. DOI: https://doi.org/10.4161/21565562.2014.988090

Vandel, A., Bouillon, M., 1959. Le Protée et son interet biologique. Ann. Speleol., 14, 111-127.

Vandel, A., 1965. Biospeleology, the biology of cavernicolous animals. Pregamon Press, Oxford, pp. 552. DOI: https://doi.org/10.1016/B978-0-08-010242-9.50027-X

Voituron, Y., de Fraipont M., Issartel, J., Guillaume, O., Clobert, J., 2011. Extreme lifespan of the human fish (Proteus anguinus): a challenge for ageing mechanisms. Biology letters, 7(1), 105-7. DOI: https://doi.org/10.1098/rsbl.2010.0539

Wake, M. H., 1968. Evolutionary morphology of the caecilian urogenital system. I. The gonads and the fat bodies. J. Morphol., 126(3), 291-331. DOI: https://doi.org/10.1002/jmor.1051260303

Downloads

Published

01.07.2017

Issue

Section

Original Research Paper

How to Cite

Bizjak Mali, L. (2017). Variability of testes morphology and the presence of testis-ova in the European blind cave salamander (Proteus anguinus). Acta Biologica Slovenica, 60(1), 53-74. https://doi.org/10.14720/abs.60.1.15669