Comparative Histological Observation of Liver Tissue Before and After Reproduction in Male and Female Frogs (Pelophylax ridibundus)

Abstract

Histomorphological structure of the liver of Ranidae species during the reproduction period contribute significantly to the production rate in the frog culture studies. In this study, the cytoplasmic status of hepatocytes, lipid and melanin pigment in liver parenchyma of adult male and female Pelophylax ridibundus (Pallas, 1771) were investigated before and after reproduction. For histological studies, adult frogs were collected from nature and placed in production ponds to examine the reproduction process. In this study, twenty four breeding male frogs, with average weight 36.63±12.84 g and length 69.29±9.15 mm, and twenty four breeding females with average weight and length of 56.61±19.59 g and 79.54±7.07 mm were used. Samples from the liver tissue of frog were taken in 2015 during (March) and after reproduction (June) period. Histologically pictures of liver parenchyma were taken from the as homogeneous fields as possible. Before the reproduction, it was found that the liver parenchyma of the female frogs were cytoplasmically intense, with a low amount of melanin pigment and lipid droplets. Similarly, male frogs liver parenchyma and hepatocytes were found to be cytoplasmically intense, containing a low amount of melanin pigment and lipid droplets. Both female and male frogs completed the reproduction in April and May. After reproduction period, a great amount of melanin pigment in the liver parenchyma was observed and the cytoplasmic density of the hepatocytes decreased. As a result, cytoplasmic decrease in hepatocytes, structural changes and increase in melanin pigments supports the idea that glycogen and lipids stored in the liver parenchyma are used for reproduction.

Keywords

• Ranidae,liver tissue,hepatocyte,histology,reproduction,Pelophylax ridibundus,reproduction

References

1. Akiyoshi, H. & Inoue, A. M. (2012). Comparative histological study of hepatic architecture in the three orders amphibian livers. Comparative Hepatology, 11(1), p. 1-8.
2. Arauco, L. R. R., De Stéfani, M. V., Nakaghi, L. S. O. & Oliveira-Bahia, V. R. L. D. (2007). Histology of kidney, liver and intestine of bullfrog tadpoles (Rana catesbeiana) fed with diets containing propolis. Ciência Rural, 37(5), 1436-1441.
3. Assisi, L., Autuori, F., Botte, V., Farrace, M. G. & Piacentini, M. (1999). Hormonal control of “tissue” transglutaminase induction during programmed cell death in frog liver. Experimental Cell Research, 247(2), 339-346.
4. Bambozzi, A. C., Seixas-Filho, J. D., Thomaz, L. A., Oshiro, L. M. Y., Braga, L. G. T. & Lima, S. L. (2004). Efeito do fotoperíodo sobre o desenvolvimento de girinos de rã-touro (Rana catesbeiana Shaw, 1802). Revista Brasileira de Zootecnia, 33(1), 1-7.
5. Barni, S. & Bernocchi, G., (1991). Internalization of erythrocytes into liver parenchymal cells in naturally hibernating frogs (Rana esculenta L.). J. Exp. Zool., 258(2), 143-150.
6. Barni, S., Bertone, V., Croce, A. C., Bottiroli, G., Bernini, F. & Gerzeli, G. (1999). Increase in liver pigmentation during natural hibernation in some amphibians. The Journal of Anatomy, 195(1), 19-25.
7. Barni, S., Vaccarone, R., Bertone, V., Fraschini, A., Bernini, F. & Fenoglio, C. (2002). Mechanisms of changes to the liver pigmentary component during the annual cycle (activity and hibernation) of Rana esculenta. L. Journal of Anatomy, 200(2), 185-194.
8. Cadeddu, G. & Castellano, S. (2012). Factors affecting variation in the reproductive investment of female treefrogs, Hyla intermedia. Zoology, 115(6), 372-378.

9. Cayuela, H., Besnard, A., Bonnaire, E., Perret, H., Rivoalen, J., Miaud, C. & Joly, P. (2014). To breed or not to breed: past reproductive status and environmental cues drive current breeding decisions in a long-lived amphibian. Oecologia, 176(1), 107-116.
10. Chen, W., Zhang, L-X. & Lu, X. (2011). Higher pre-hibernation energy storage in anurans from cold environments: A case study on a temperate frog Rana chensinensis along a broad latitudinal and altitudinal gradients. Annales Zoologici Fennici, 48(4), 214–220.
11. Crawshaw, G.J. & Weinkle, T.K. (2000). Clinical and pathological aspects of the amphibian liver. Seminars in Avian and Exotic Pet Medicine, 9(3), 165-173.
12. Dinsmore, S. C. & Swanson, D. L. (2008). Temporal patterns of tissue glycogen, glucose, and glycogen phosphorylase activity prior to hibernation in freeze-tolerant chorus frogs, Pseudacris triseriata. Canadian Journal of Zoology, 86(10), 1095-1100.
13. Fenoglio, C., Bernocchi, G. & Barni, S. (1992). Frog hepatocyte modifications induced by seasonal variations: a morphological and cytochemical study. Tissue and Cell, 24(1), 17-29.
14. Gernhofer, M., Pawet, M., Schramm, M., Müller, E. & Triebskorn, R. (2001). Ultrastructural Biomarkers as Tools to Characterize the Health Status of Fish in Contaminated Streams. Journal of Aquatic Ecosystem Stress and Recovery, 8, 241-226.
15. Haar, J. L. & Hightower, J.A.A. (1976). Light and electron microscopic investigation of the hepatic parenchyma of the adult newt, Notophthalmus viridescens. The Anatomical Record, 185(3), 313-323.
16. Hipolito, M., Leme, M. C. M. & Bach, E. E. (2001). Lesões anátomo-histopatológicas em rãs-touro (Rana catesbeiana Shaw, 1802) associadas à deterioração da ração. Arquivos do Instituto de Biologia, 68(1), 111-114.
17. Hipolito, M., Martins, A. M. C. R. P. F. & Bach, E. E. (2004). Aspectos bioquímicos em fígado de rãs-touro (Rana catesbeiana Shaw, 1802) sadias e doentes. Arquivos do Instituto Biológico, 71(2), 147-153.
18. Mentino, D., Mastrodonato, M., Rossi, R. & Scillitani, G., (2014). Histochemical and structural characterization of egg extra-cellular matrix in bufonid toads, Bufo bufo and Bufotes balearicus: Molecular diversity versus morphological uniformity. Microscopy Research and Technique, 77(11), 910–917.
19. Mentino, D., Scillitani, G., Marra, M. & Mastrodonato, M. (2017). Seasonal changes in the liver of a non-hibernating population of water frogs, Pelophylax kl. esculentus (Anura: Ranidae). The European Zoological Journal, 84(1), 525-535.
20. Pasanen, S. & Koskela, P. (1974). Seasonal and age variation in the metabolism of the common frog, Rana temporaria L. in northern Finland. Comparative Biochemistry and Physiology Part A: Physiology, 47(2), 635-654.
21. Purrello, M., Scalia, M., Corsaro, C., Di Pietro, C., Piro, S. & Sichel, G. (2001). Melanosynthesis, differentiation, and apoptosis in Kupffer cells from Rana esculenta. Pigment cell research, 14(2), 126-131.
22. Seixas Filho, J. T., Hipólito, M., Pereira, M. M., Martins, A. M. C. R. P. F., Rodrigues, E. and Mello, S.C.R.P., (2013). Liver histopathological changes in breeding bullfrogs. Acta Scientiarium Animal Science, 35:461-465.
23. Singh, R. P. & Sinha, R. C. (1989). Seasonal changes in energy reserves in the common frog, Rana tigrina. The Japanese Journal of Physiology, 39(6), 969-973.
24. Wester, P. W. & Canton, J. H. (1991). The usefulness of histopathology in aquatic toxicity studies. Comparative Biochemistry and Physiology. C, Comparative Pharmacology and Toxicology, 100(1-2), 115-117.