Persulfidation contributes to a group of redox post-translational modifications (PTMs), which arise exclusively on the sulfhydryl group of cysteine as a result of hydrogen sulfide (H2S) action. Redox-active molecules, including H2S, contribute to sperm development; therefore, redox PTMs represent an extremely important signalling pathway in sperm life. In this path, persulfidation prevents protein damage caused by irreversible cysteine hyperoxidation and thus maintains this signalling pathway. In our study, we detected both H2S and its production by all H2S-releasing enzymes (cystathionine γ-lyase (CTH), cystathionine β-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (MPST)) in male reproduction, including spermatozoa. We provided evidence that sperm H2S leads to persulfidation of proteins, such as glyceraldehyde-3-phosphate dehydrogenase, tubulin, and anchor protein A-kinase. Overall, this study suggests that persulfidation, as a part of the redox signalling pathway, is tightly regulated by enzymatic H2S production and is required for sperm viability.
- MeSH
- cystathionin-gama-lyasa metabolismus MeSH
- cystein metabolismus MeSH
- lidé MeSH
- rozmnožování MeSH
- sperma metabolismus MeSH
- sulfan * metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
The aim of this study was to evaluate the mutual relationship among perivascular adipose tissue (PVAT) and endogenous and exogenous H2S in vasoactive responses of isolated arteries from adult normotensive (Wistar) rats and hypertriglyceridemic (HTG) rats, which are a nonobese model of metabolic syndrome. In HTG rats, mild hypertension was associated with glucose intolerance, dyslipidemia, increased amount of retroperitoneal fat, increased arterial contractility, and endothelial dysfunction associated with arterial wall injury, which was accompanied by decreased nitric oxide (NO)-synthase activity, increased expression of H2S producing enzyme, and an altered oxidative state. In HTG, endogenous H2S participated in the inhibition of endothelium-dependent vasorelaxation regardless of PVAT presence; on the other hand, aortas with preserved PVAT revealed a stronger anticontractile effect mediated at least partially by H2S. Although we observed a higher vasorelaxation induced by exogenous H2S donor in HTG rats than in Wistar rats, intact PVAT subtilized this effect. We demonstrate that, in HTG rats, endogenous H2S could manifest a dual effect depending on the type of triggered signaling pathway. H2S within the arterial wall contributes to endothelial dysfunction. On the other hand, PVAT of HTG is endowed with compensatory vasoactive mechanisms, which include stronger anti-contractile action of H2S. Nevertheless, the possible negative impact of PVAT during hypertriglyceridemia on the activity of exogenous H2S donors needs to be taken into consideration.
- MeSH
- aorta thoracica patofyziologie MeSH
- cévní endotel patofyziologie MeSH
- cystathionin-gama-lyasa metabolismus MeSH
- hypertriglyceridemie metabolismus MeSH
- metabolický syndrom metabolismus patofyziologie MeSH
- modely nemocí na zvířatech MeSH
- noradrenalin farmakologie MeSH
- oxidace-redukce MeSH
- potkani Wistar MeSH
- signální transdukce * MeSH
- superoxiddismutasa metabolismus MeSH
- superoxidy metabolismus MeSH
- synthasa oxidu dusnatého, typ III metabolismus MeSH
- tuková tkáň metabolismus MeSH
- vazodilatace fyziologie MeSH
- vazokonstrikce účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The role of hydrogen sulfide (H2S) is addressed in Xenopuslaevis oocytes. Three enzymes involved in H2S metabolism, cystathionine β-synthase, cystathionine γ-lyase, and 3-mercaptopyruvate sulfurtransferase, were detected in prophase I and metaphase II-arrested oocytes and drove an acceleration of oocyte meiosis resumption when inhibited. Moreover, meiosis resumption is associated with a significant decrease in endogenous H2S. On another hand, a dose-dependent inhibition was obtained using the H2S donor, NaHS (1 and 5 mM). NaHS impaired translation. NaHS did not induce the dissociation of the components of the M-phase promoting factor (MPF), cyclin B and Cdk1, nor directly impacted the MPF activity. However, the M-phase entry induced by microinjection of metaphase II MPF-containing cytoplasm was diminished, suggesting upstream components of the MPF auto-amplification loop were sensitive to H2S. Superoxide dismutase and catalase hindered the effects of NaHS, and this sensitivity was partially dependent on the production of reactive oxygen species (ROS). In contrast to other species, no apoptosis was promoted. These results suggest a contribution of H2S signaling in the timing of amphibian oocytes meiosis resumption.
- MeSH
- apoptóza účinky léků MeSH
- cyklin B metabolismus MeSH
- cystathionin-beta-synthasa antagonisté a inhibitory metabolismus MeSH
- cystathionin-gama-lyasa antagonisté a inhibitory metabolismus MeSH
- cytoplazma metabolismus MeSH
- faktor podporující zrání metabolismus MeSH
- fosfatasy cdc25 metabolismus MeSH
- katalasa metabolismus MeSH
- meióza účinky léků MeSH
- metafáze účinky léků MeSH
- oocyty chemie enzymologie metabolismus MeSH
- profáze meiózy I účinky léků MeSH
- proteinkinasy metabolismus MeSH
- proteiny buněčného cyklu metabolismus MeSH
- proteiny Xenopus metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- signální transdukce účinky léků MeSH
- sulfan metabolismus MeSH
- sulfidy metabolismus farmakologie MeSH
- sulfurtransferasy antagonisté a inhibitory metabolismus MeSH
- superoxiddismutasa metabolismus MeSH
- viabilita buněk účinky léků MeSH
- Xenopus laevis MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Knowledge about the expression and thus a role of enzymes that produce endogenous H2S - cystathionine-β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase - in renal tumors is still controversial. In this study we aimed to determine the expression of these enzymes relatively to the expression in unaffected part of kidney from the same patient and to found relation of these changes to apoptosis. To evaluate patient's samples, microarray and immunohistochemistry was used. METHODS: To determine the physiological importance, we used RCC4 stable cell line derived from clear cell renal cell carcinoma, where apoptosis induction by a mixture of five chemotherapeutics with/without silencing of H2S-producing enzymes was detected. Immunofluorescence was used to determine each enzyme in the cells. RESULTS: In clear cell renal cell carcinomas, expression of H2S-producing enzymes was mostly decreased compared to a part of kidney that was distal from the tumor. To evaluate a potential role of H2S-producing enzymes in the apoptosis induction, we used RCC4 stable cell line. We have found that silencing of cystathionine-β-synthase and cystathionine γ-lyase prevented induction of apoptosis. Immunofluorescence staining clearly showed that these enzymes were upregulated during apoptosis in RCC4 cells. CONCLUSION: Based on these results we concluded that in clear cell renal cell carcinoma, reduced expression of the H2S-producing enzymes, mainly cystathionine γ-lyase, might contribute to a resistance to the induction of apoptosis. Increased production of the endogenous H2S, or donation from the external sources might be of a therapeutic importance in these tumors.
- MeSH
- apoptóza * MeSH
- cystathionin-beta-synthasa genetika metabolismus MeSH
- cystathionin-gama-lyasa genetika metabolismus MeSH
- dospělí MeSH
- karcinom z renálních buněk patologie chirurgie MeSH
- ledviny metabolismus patologie chirurgie MeSH
- lidé středního věku MeSH
- lidé MeSH
- malá interferující RNA metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory ledvin patologie chirurgie MeSH
- nefrektomie MeSH
- RNA interference MeSH
- senioři MeSH
- sulfan metabolismus MeSH
- upregulace MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Porcine oocytes that have matured in in vitro conditions undergo the process of aging during prolonged cultivation, which is manifested by spontaneous parthenogenetic activation, lysis or fragmentation of aged oocytes. This study focused on the role of hydrogen sulfide (H2S) in the process of porcine oocyte aging. H2S is a gaseous signaling molecule and is produced endogenously by the enzymes cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (MPST). We demonstrated that H2S-producing enzymes are active in porcine oocytes and that a statistically significant decline in endogenous H2S production occurs during the first day of aging. Inhibition of these enzymes accelerates signs of aging in oocytes and significantly increases the ratio of fragmented oocytes. The presence of exogenous H2S from a donor (Na2S.9H2O) significantly suppressed the manifestations of aging, reversed the effects of inhibitors and resulted in the complete suppression of oocyte fragmentation. Cultivation of aging oocytes in the presence of H2S donor positively affected their subsequent embryonic development following parthenogenetic activation. Although no unambiguous effects of exogenous H2S on MPF and MAPK activities were detected and the intracellular mechanism underlying H2S activity remains unclear, our study clearly demonstrates the role of H2S in the regulation of porcine oocyte aging.
- MeSH
- cystathionin-beta-synthasa metabolismus MeSH
- cystathionin-gama-lyasa metabolismus MeSH
- embryo savčí účinky léků MeSH
- inhibitory enzymů farmakologie MeSH
- kultivace embrya MeSH
- kultivované buňky MeSH
- oocyty účinky léků fyziologie MeSH
- partenogeneze účinky léků MeSH
- prasata MeSH
- stárnutí buněk účinky léků MeSH
- sulfan metabolismus farmakologie MeSH
- sulfurtransferasy metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hereditary cystathioninuria is due to mutations in the CTH gene that encodes for cystathionase, a pyridoxal-5'-phosphate (PLP) dependent enzyme. To date, mutations in this gene have been described in 10 unrelated cystathioninuric patients. Enzyme assays have showed that mutated cystathionase exhibits lower activity than controls. As cystathioninuria is usually accompanied by a wide variety of symptoms, it has been questioned whether it is a disease or just a biochemical finding not associated with the clinical picture of these patients. This is the first report of Spanish patients with cystathioninuria and mild to severe neurological symptoms in childhood. After oral pyridoxine therapy biochemical parameters have normalized but clinical amelioration was not evident. All patients were homozygotes for the c.200C>T (p.T67I) variant which is the most prevalent inactivating mutation in the CTH gene. To further investigate the history of the alleles carrying the c.200C>T transition in Europe, we also constructed the haplotypes on the CTH locus in our Spanish patients as well as in a clinical series of cystathioninuric patients from the Czech Republic harboring the same nucleotide change. We suggest that the CTH p.T67I substitution could have an ancient common origin, which probably occurred in the Neolithic Era and spread throughout Europe.
- MeSH
- alely * MeSH
- cystathionin-gama-lyasa genetika MeSH
- dítě MeSH
- genetická variace genetika MeSH
- hyperhomocysteinemie genetika MeSH
- lidé MeSH
- předškolní dítě MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Evropa MeSH
O sirovodíku (H2S) bylo prokázáno, že je důležitou signální biologickou molekulou endogenního původu. V savčích tkáních je produkován endogenně zejména z cysteinu třemi různými enzymy a hraje důležitou úlohu za fyziologických a patofyziologických podmínek v centrální nervové soustavě, v kardiovaskulárním systému a v hladkém svalstvu. Pro výzkum je také inspirující účinek sirovodíku snižující metabolický obrat a navozující u experimantálních hlodavců stav blízký hibernaci.
Hydrogen sulfide (H2S) has been shown to be an important signaling biological molecule of endogenous origin. It is produced endogenously especially from cystein by three enzymes in mammalian tissues and plays important roles in physiological and pathophysiological conditions in the central nervous system, in cardiovascular system and in smooth muscles. A challange for research is the effect of hydrogen sulphide inducing hypometabolism and mimeting hibernation.
- MeSH
- cystathionin-beta-synthasa metabolismus MeSH
- cystathionin-gama-lyasa metabolismus MeSH
- hibernace MeSH
- kardiovaskulární systém enzymologie chemie MeSH
- modely u zvířat MeSH
- neurotransmiterové látky fyziologie MeSH
- střevní nervový systém chemie MeSH
- sulfan farmakokinetika farmakologie metabolismus MeSH
Hydrogen sulfide is an important signalling biological compound of endogenous origin. It is produced in mammalian tissues by enzymes ß-synthase and cystathionine ?-lyase especially from cysteine and homocysteine. H2S plays important roles under physiological and pathophysiological conditions in the central nervous and cardiovascular systems, in smooth muscles and other organs.
- MeSH
- centrální nervový systém chemie účinky léků MeSH
- cystathionin-beta-synthasa metabolismus MeSH
- cystathionin-gama-lyasa metabolismus MeSH
- financování organizované MeSH
- hormony farmakokinetika farmakologie chemie MeSH
- kardiovaskulární systém chemie účinky léků MeSH
- lidé MeSH
- metabolismus fyziologie účinky léků MeSH
- plyny farmakologie chemie izolace a purifikace MeSH
- sulfan farmakokinetika farmakologie chemie MeSH
- svaly chemie účinky léků MeSH
- zánět metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH