The hypothalamic suprachiasmatic nuclei (SCN), the circadian master clock in mammals, releases ATP in a rhythm, but the role of extracellular ATP in the SCN is still unknown. In this study, we examined the expression and function of ATP-gated P2X receptors (P2XRs) in the SCN neurons of slices isolated from the brain of 16- to 20-day-old rats. Quantitative RT-PCR showed that the SCN contains mRNA for P2X 1-7 receptors and several G-protein-coupled P2Y receptors. Among the P2XR subunits, the P2X2 > P2X7 > P2X4 mRNAs were the most abundant. Whole-cell patch-clamp recordings from SCN neurons revealed that extracellular ATP application increased the frequency of spontaneous GABAergic IPSCs without changes in their amplitudes. The effect of ATP appears to be mediated by presynaptic P2X2Rs because ATPγS and 2MeS-ATP mimics, while the P2XR antagonist PPADS blocks, the observed enhancement of the frequency of GABA currents. There were significant differences between two SCN regions in that the effect of ATP was higher in the ventrolateral subdivision, which is densely innervated from outside the SCN. Little evidence was found for the presence of P2XR channels in somata of SCN neurons as P2X2R immunoreactivity colocalized with synapsin and ATP-induced current was observed in only 7% of cells. In fura-2 AM-loaded slices, BzATP as well as ADP stimulated intracellular Ca(2+) increase, indicating that the SCN cells express functional P2X7 and P2Y receptors. Our data suggest that ATP activates presynaptic P2X2Rs to regulate inhibitory synaptic transmission within the SCN and that this effect varies between regions.

• MeSH
• Adenosine Triphosphate pharmacology   MeSH
• Excitatory Amino Acid Antagonists pharmacology   MeSH
• Biophysical Phenomena drug effects   MeSH
• Sodium Channel Blockers pharmacology   MeSH
• gamma-Aminobutyric Acid pharmacology   MeSH
• Platelet Aggregation Inhibitors pharmacology   MeSH
• Rats MeSH
• Cells, Cultured MeSH
• RNA, Messenger metabolism   MeSH
• Patch-Clamp Techniques MeSH
• Synaptic Transmission drug effects   MeSH
• Neural Inhibition drug effects   MeSH
• Neurons drug effects   MeSH
• Animals, Newborn MeSH
• Suprachiasmatic Nucleus cytology   MeSH
• Rats, Wistar MeSH
• Purinergic Agents pharmacology   MeSH
• Receptors, Purinergic P2X genetics   metabolism   MeSH
• Gene Expression Regulation drug effects   MeSH
• Synaptic Potentials drug effects   MeSH
• In Vitro Techniques MeSH
• Tetrodotoxin pharmacology   MeSH
• Calcium metabolism   MeSH
• Dose-Response Relationship, Drug MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Adenosine Triphosphate MeSH
• Excitatory Amino Acid Antagonists MeSH
• Sodium Channel Blockers MeSH
• gamma-Aminobutyric Acid MeSH
• Platelet Aggregation Inhibitors MeSH
• RNA, Messenger MeSH
• Purinergic Agents MeSH
• Receptors, Purinergic P2X MeSH
• Tetrodotoxin MeSH
• Calcium MeSH

In spite of the inhibitory effects of ethanol (EtOH) on platelet function, soft blood clots are often observed in cadaveric blood in cases of sudden death after alcohol ingestion. In order to resolve this discrepancy, we have focused on the role of vascular endothelial cells. We tried to investigate the effects of EtOH and LPS on endothelial cells from various perspectives; thrombogenic factor (Von Willebrand factor, VWF), fibrinolytic factor (tissue plasminogen activator, tPA) and inflammatory factor (Interleukin-6, IL-6). Human umbilical vein endothelial cells (HUVECs) were incubated with various concentrations of EtOH (0-160 mM) with or without LPS. Treatment with EtOH and LPS increased VWF release from HUVECs without enhancement mRNA expression. Treatment with 40 mM of EtOH also increased IL-6 release from HUVECs without enhancement mRNA expression. Although EtOH inhibited LPS-induced IL-6 mRNA expression, 20 mM of EtOH still had an increasing effect on the release of IL-6. These doses of EtOH are consistent with a moderate drunkenness level in a normal person. On the other hand, mRNA expression and release reaction of tPA were not affected by EtOH and LPS addition. In conclusion, EtOH enhances procoagulant status via VWF release and IL-6 production cooperation with LPS and may contribute to soft blood clot formation in cadaveric blood.

• MeSH
• Cell Line MeSH
• Endothelium, Vascular drug effects   metabolism   MeSH
• Ethanol pharmacology   MeSH
• Blood Coagulation MeSH
• Interleukin-6 metabolism   MeSH
• Blood Coagulation Factors metabolism   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Lipopolysaccharides pharmacology   MeSH
• Postmortem Changes MeSH
• Tissue Plasminogen Activator metabolism   MeSH
• Umbilical Veins drug effects   MeSH
• von Willebrand Factor metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Ethanol MeSH
• Interleukin-6 MeSH
• Blood Coagulation Factors MeSH
• Lipopolysaccharides MeSH
• Tissue Plasminogen Activator MeSH
• von Willebrand Factor MeSH

The aim of present study was to evaluate the action of green tea and its constituents on rabbit ovarian functions and some non-reproductive indexes. In in vitro experiments, rabbit ovarian fragments were cultured with green tea constituents - epigallocatechin-3-gallate (EGCG), green tea polyphenols (GTPP) and resveratrol (RSV) (at 0, 1, 10 or 100 μg/mL medium). The accumulation of an apoptosis marker - caspase 3 and the release of progesterone (P4) and testosterone (T) were measured. In in vivo experiments, does were fed a standard diet or a diet enriched with green tea powder. The weight gain, mortality, ovarian length and weight, conception and kindling rate, number of liveborn, stillborn, and weaned pups, diameter of ovarian follicles and some blood haematological and biochemical parameters were analysed. Culture of ovarian fragments with EGCG increased accumulation of caspase 3, whilst both GTTP and RSV decreased it. EGCG inhibited both P4 and T output, GTPP stimulated P4 and inhibited T, whilst RSV promoted release of both P4 and T. Feeding with green tea increased ovarian length and diameter of ovarian non-ovulated peri-ovulatory haemorrhagic but not of primary and secondary growing follicles. Furthermore, green tea reduced conception and kindling rate, the number of liveborn and weaned pups, increased female mortality but not their weight gain. It reduced platelet distribution width, but it did not affect other haematological and biochemical indexes. These observations suggest that dietary green tea can reduce rabbit doe's viability, ovarian functions and fecundity, perhaps due to changes in ovarian cell apoptosis, steroid hormones release and blockade of the ovulation of large ovarian follicles. The anti-reproductive action of green tea could be due to its constituent - EGCG with pro-apoptotic and anti-steroid hormone properties.

• Keywords
• Conception rate, Fecundity, Green tea, Ovaries histology, Rabbit female, Steroid hormones,
• MeSH
• Apoptosis MeSH
• Tea chemistry   MeSH
• Fertility drug effects   MeSH
• Caspase 3 metabolism   MeSH
• Catechin analogs & derivatives   isolation & purification   pharmacology   MeSH
• Rabbits * MeSH
• Ovary drug effects   metabolism   MeSH
• Polyphenols isolation & purification   pharmacology   MeSH
• Resveratrol isolation & purification   pharmacology   MeSH
• Animals MeSH
• Check Tag
• Rabbits * MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Tea MeSH
• epigallocatechin gallate MeSH Browser
• Caspase 3 MeSH
• Catechin MeSH
• Polyphenols MeSH
• Resveratrol MeSH