Laboratory of Reproductive Physiology
University of South Bohemia in České Budějovice
Faculty of Fisheries and Protection of Waters
Research Institute of Fish Culture and Hydrobiology
Contact
Prof. Otomar Linhart (dean of faculty)
linhart@frov.jcu.cz
Phone: +420 38777 4600
Dr. Martin Pšenička (head of laboratory, coordinator of STSMs)
psenicka@frov.jcu.cz
Phone: +420 725 787 925
Address
University of South Bohemia in České Budějovice
Faculty of Fisheries and Protection of Waters
Research Institute of Fish Culture and Hydrobiology
Zátiší 728/II, 389 25 Vodňany
Link
www.frov.jcu.cz
Group description
The laboratory carries out besides lecturing also basic and applied research with focus on reproduction of our economically significant and endangered fish species, especially sturgeons. The main tools are microscopes equipped with stroboscopic lamp, high speed camera and computer assisted system and software of image analyses for evaluation of sperm motility parameters, bioluminescent method for ATP measuring, proteomics, micromanipulations, fluorescent, confocal and electron microscopy. Tests of fertilization and hatching are performed at Genetic fisheries centre. The outputs are in form of methodologies, technologies and research articles focused on activation, reactivation and immobilization of gametes, sex determination, determination of maturity stages in females, gen and genomic manipulations, short and long term storage of gametes (cryoconservation), sex reversal and evaluation of endocrine disruptor effects. Technics of germ cells micromanipulations are used for cryopreservation of endangered and important fish species, for interspecies transplantations between species with different length of sex maturation, etc. The laboratory possesses a gen bank with cryopreserved sperm, blastomers and spermatogonia of various fish species and breeds.
The laboratory also offers assistance in sex and maturity stage determination and in artificial reproduction of fish itself.
The description of rotation of sterlet spermatozoa flagella on the bottom of drop. Time between frames is 2 millisec. This picture is from the study of Sergey Boryshpolets, which gained the award for the best presentation of PhD student during the 3rd International Workshop on the Biology of Fish Gametes in Budapest, Hungary.
Boryshpolets, S., Cosson, J., Bondarenko, V., Gillies, E., Rodina, M., Dzyuba, B. and Linhart O. Different swimming behaviours of sterlet (Acipenser ruthenus) spermatozoa close to solid and free surfaces. Theriogenology 79(1):81‐6.
The acrosome of Siberian sturgeon before and after activation.
Psenicka, M., Rodina, M., Linhart, O., 2010. Ultrastructural study on fertilization process in sturgeon
(Acipenser), function of acrosome and prevention of polyspermy. Animal Reproduction Science 117: 147–154.
Freezing of fish sperm and germ cells
A) Work on freezing machine freezing;
B) Freezing machine for cell freezing to -196 °C;
C) Gutters with individual boxes, heating, recirculation and UV sterilization are used for testing fertilization and hatching performance.
The determination of sex and oocyte stage using biopsy and collection of eggs in sterlet.
The vizualization of sturgeon (A.ruthenus) primordial germ cells and their transplantation into egg of Xenopus laevis.
The hermaphroditic carp gonad with spermatocytes and oocyte.
Relevant publications
1. Hatef, A., Alavi, S.M.H., Golshan, M., Linhart, O., 2013. Toxicity of environmental contaminants to fish spermatozoa function in vitro—A review. Aquatic Toxicology 140– 141: 134–144.
2. Li, P., Hulák, M., Li, Z.H., Šulc, M., Pšenička, M., Rodina, M., Gela, D., Linhart, O., 2013. Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein hosphorylation in tyrosine and threonine residues. Theriogenology 80: 84–89.
3. Dzyuba, B., Cosson, J., Yamener, G., Bondarenko, O., Rodina, M., Gela, D., Bondarenko, V., Shaliutina, A., Linhart, O., 2013. Hypotonic treatment prior to freezing improves cryoresistance of common carp (Cyprinus carpio L.) spermatozoa. Cryobiology 66 (2): 192–194.
4. Boryshpolets, S., Cosson, J., Bondarenko V., Gillies, Rodina, M., Dzyuba, B., Linhart, O., 2013. Different swimming behaviors of sterlet (Acipenser ruthenus) spermatozoa close to solid and free surfaces. Theriogenology 79: 81–86.
5. Bondarenko, O., Dzyuba, B., Cosson, J., Yamanater, G., Prokopchuk, G., Pšenička, M., Linhart, O., 2013. Volume changes during the motility period of fish spermatozoa: Interspecies differences. Theriogenology 79 (5): 872–881.
6. Shaliutina, A., Hulák, M., Gazo, I., Linhartová, P., Linhart, O., 2013. Effect of short-term storage on quality parameters, DNA integrity, and oxidative stress in Russian (Acipenser gueldenstaedtii) and Siberian (Acipenser baerii) sturgeon sperm. Animal Reproduction Science 139: 127–135.
7. Alavi, S.M.H., Rodina, M., Gela, D., Linhart, O., 2012. Sperm biology and control of reproduction in sturgeon: (I) testicular development, sperm maturation and seminal plasma characteristics. Rev Fish Biol Fisheries 22: 695-717.
8. Alavi, S.M.H., Hatef, A., Pšenička, M., Kašpar, V., Boryshpolets, S., Dzyuba, B., Cosson, J., Bondarenko, V., Rodina, M., Gela, D., Linhart, O., 2012. Sperm biology and control of reproduction in sturgeon: (II) Sperm morphology, acrosome reaction, motility and cryopreservation. Reviews in Fish Biology and Fisheries 22: 861–886.
9. Dzyuba, B., Boryshpolets, S., Shaliutina, A., Rodina, M., Yamaner, G., Gela, D., Dzyuba, V., Linhart, O., 2012. Spermatozoa motility, cryoresistance, and fertilizing ability in sterlet Acipenser ruthenus during sequential stripping. Aquaculture 356-357: 272-278.
10. Dzyuba, B., Cosson, J., Yamaner, G., Bondarenko, O., Rodina, M., Gela, D., Bondarenko, V., Shaliutina, A., Linhart, O., 2012. Hypotonic treatment prior to freezing improves cryoresistance of common carp (Cyprinus carpio L.) spermatozoa. Cryobiology 66(2):192-4.
11. Hatef, A., Alavi, S.M.H., Abdulfatah, A., Fontaine, P., Rodina, M., Linhart, O., 2012. Adverse effects of Bisphenol A on reproductive physiology in male goldfish at environmentally relevant concentrations. Ecotoxicology and Environmental Safety 76: 56-62. (IF 2011 = 2,294)
12. Hatef, A., Alavi, S.M.H., Milla, S., Křišťan, J., Golshan, M., Fontaine, P., Linhart, O., 2012. Anti-androgen vinclozolin impairs sperm quality and steroidogenesis in goldfish. Aquatic Toxicology 122–123: 181–187.
13. Li, P., Li, Z.H., Dzyuba, B., Hulak, M., Rodina, M., Linhart O., 2010. Evaluating the impacts of osmotic and oxidative stress on common carp (Cyprinus carpio, L.) sperm caused by cryopreservation techniques. Biology of Reproduction 83: 852-858.
14. Psenicka, M., Rodina, M., Linhart, O., 2010. Ultrastructural study on fertilization process in sturgeon (Acipenser), function of acrosome and prevention of polyspermy. Animal Reproduction Science 117: 147-154.
15. Pšenička, M., Flajšhans, M., Hulák, M., Kašpar, V., Rodina, M., Borishpolet, S., Linhart, O., 2010. The influence of ploidy level on ultrastructure and motility of tench (Tinca tinca L.) spermatozoa. Reviews in Fish Biology and Fisheries 20 (3): 331-338.