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Demembranation of fish sperm: Design and verification this procedure for the different species of freshwaterfish and demonstration usage of this technique by study the effect of heavy metals to sperm axoneme
BLAŽKOVÁ, Jaroslava
The object of this study is to design demembranation method on four freshwater species and its application on study of the influence of HgCl2 on the axoneme and motity sperm motility parameters. Demembranation was designed and examined for all investigated species common carp (Cyprinus carpio), sterlet (Acipenser ruthenus), perch (Perca fluviatilis) and african catfish (Clarias gariepinus). One-step and two-step method was designed and tested for common carp. One-step method was designed for sterlet and perch. Two-step method of demembranation was designed for african catfish. Demembranation was designed and examined for all species under examination. Sperm motility was evidently increased above normal physiological value. Other sperm motility parameters (velocity, percent of motile cells) slightly decreased. HgCl2 in concentration 0,01 mM to the demembranation medium didn't show effect on flagellar microtubule aparat and then to the motility parameters, except sterlet; demembranated sterlet sperm was inhibited at all used concentration of HgCl2. Concentration 0,1 mM had inhibition effect on carp and africant catfish spermatozoa. Concentration 1 mM HgCl2 inhibited sperm of all tested species.
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Vliv endokrinních disruptorů na funkčnost spermií a změny ve steroidogenezi transkriptomik u ryb
HATEF, Azadeh
In the present study, both in vitro and in vivo experiments were performed to investigate modes of action of selected EDCs including mercury chloride (HgCl2), bisphenol A (BPA), vinclozolin (VZ) and Di-(2-ethylhexyl)-phthalate (DEHP) on male reproductive physiology in fish. Results obtained from in vitro studies showed that EDCs decrease sperm motility via disruption in energetics and damage to spermatozoa; however the effective doses are higher than reported in the environmentally relevant concentrations. In contrast, in vivo studies showed adverse effects of BPA, VZ, and DEHP on sperm quality at environmentally relevant concentrations. Decrease in sperm quality is associated with disruption in testicular steroidogenesis, particularly inhibition of androgens. Only BPA induced vitellogenin production, associated with increase in transcripts of estrogen receptor, androgen receptor and cytochrome P450 mRNA at the high dose approaching environmental level. Inhibition of androgen production is due to disruption in transfer of cholesterol into the steroidogenesis pathway, because transcript of steroidogenic acute regulatory protein mRNA was decreased. In addition, BPA acts through sex steroid-mediated mechanism while DEHP mode of action is sex-steroid independent mechanism.
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