tacrolimus and propargylglycine

Acute hypoxia depolarizes carotid body chemoreceptor (glomus) cells and elevates intracellular Ca(2+) concentration ([Ca(2+)]i). Recent studies suggest that hydrogen sulfide (H2S) may serve as an oxygen sensor/signal in the carotid body during acute hypoxia. To further test such a role for H2S, we studied the effects of H2S on the activity of TASK channel and [Ca(2+)]i, which are considered important for mediating the glomus cell response to hypoxia. Like hypoxia, NaHS (a H2S donor) inhibited TASK activity and elevated [Ca(2+)]i. To inhibit the production of H2S, glomus cells were incubated (3h) with inhibitors of cystathionine-β-synthase and cystathionine-γ-lyase (DL-propargylglycine, aminooxyacetic acid, β-cyano-L-alanine; 0.3 mM). SF7 fluorescence was used to assess the level of H2S production. The inhibitors blocked L-cysteine- and hypoxia-induced elevation of SF7 fluorescence intensity. In cells treated with the inhibitors, hypoxia produced an inhibition of TASK activity and a rise in [Ca(2+)]i, similar in magnitude to those observed in control cells. L-cysteine produced no effect on TASK activity or [Ca(2+)]i and did not affect hypoxia-induced inhibition of TASK and elevation of [Ca(2+)]i. These findings suggest that under normal conditions, H2S is not a major signal in hypoxia-induced modulation of TASK channels and [Ca(2+)]i in isolated glomus cells.

Topics: Alkynes; Analysis of Variance; Animals; Animals, Newborn; Calcium; Carotid Body; Chemoreceptor Cells; Dose-Response Relationship, Drug; Gasotransmitters; Glycine; Hydrogen Sulfide; Hypoxia; Intracellular Fluid; Membrane Potentials; Nerve Tissue Proteins; Patch-Clamp Techniques; Potassium Channels, Tandem Pore Domain; Rats; Rats, Sprague-Dawley; Sodium Compounds; Tacrolimus; Time Factors

The role of hydrogen sulphide (H₂S) as a putative endogenous signalling molecule in the gastrointestinal tract has not yet been established. We investigated the effect of D,L-propargylglycine (PAG), an inhibitor of cystathionine γ-lyase (CSE), amino-oxyacetic acid (AOAA) and hydroxylamine (HA), inhibitors of cystathionine β-synthase (CBS) on rat colonic motility.. Immunohistochemistry, H₂S production, microelectrode and organ bath recordings were performed on rat colonic samples without mucosa and submucosa to investigate the role of endogenous H₂S in motility.. CSE and CBS were immunolocalized in the colon. H₂S was endogenously produced (15.6 ± 0.7 nmol·min⁻¹·g⁻¹ tissue) and its production was strongly inhibited by PAG (2 mM) and AOAA (2 mM). PAG (2 mM) caused smooth muscle depolarization and increased spontaneous motility. The effect was still recorded after incubation with tetrodotoxin (TTX, 1 µM) or N(ω) -nitro-L-arginine (L-NNA, 1 mM). AOAA (2 mM) caused a transient (10 min) increase in motility. In contrast, HA (10 µM) caused a 'nitric oxide-like effect', smooth muscle hyperpolarization and relaxation, which were antagonized by 1H-[1,2,4]oxadiazolo[4,3-α]quinoxalin-1-one (ODQ, 10 µM). Neither spontaneous nor induced inhibitory junction potentials were modified by AOAA or PAG.. We demonstrated that H₂S is endogenously produced in the rat colon. PAG and AOAA effectively blocked H₂S production. Our data suggest that enzymatic production of H₂S regulates colonic motility and therefore H₂S ight be a third gaseous inhibitory signalling molecule in the gastrointestinal tract. However, possible non-specific effects of the inhibitors should be considered.

Topics: Alkynes; Animals; Colon; Cystathionine beta-Synthase; Cystathionine gamma-Lyase; Cysteine; Enzyme Inhibitors; Gastrointestinal Motility; Gastrointestinal Tract; Glycine; Hydrogen Sulfide; Hydroxylamine; Male; Membrane Potentials; Muscle, Smooth; Nitric Oxide; Rats; Rats, Sprague-Dawley; Tacrolimus; Tetrodotoxin