ABSTRACT
After depletion of intracellular Ca2+ stores the capacitative response triggers an extracellular Ca2+ influx through store-operated channels (SOCs) which refills these stores. Our objective was to explore if human umbilical artery smooth muscle presented this response and if it was involved in the mechanism of serotonin- and histamine-induced contractions. Intracellular Ca2+ depletion by a Ca2+-free extracellular solution followed by Ca2+ readdition produced a contraction in artery rings which was inhibited by the blocker of Orai and TRPC channels 2-aminoethoxydiphenyl borate (2-APB), suggesting a capacitative response. In presence of 2-APB the magnitude of a second paired contraction by serotonin or histamine was significantly less than a first one, likely because 2-APB inhibited store refilling by capacitative Ca2+ entry. 2-APB inhibition of sarcoplasmic reticulum Ca2+ release was excluded because this blocker did not affect serotonin force development in a Ca2+-free solution. The PCR technique showed the presence of mRNAs for STIM proteins (1 and 2), for Orai proteins (1, 2 and 3) and for TRPC channels (subtypes 1, 3, 4 and 6) in the smooth muscle of the human umbilical artery. Hence, this artery presents a capacitative contractile response triggered by stimulation with physiological vasoconstrictors and expresses mRNAs for proteins and channels previously identified as SOCs.
Subject(s)
Humans , Boron Compounds/pharmacology , Muscle Contraction/drug effects , Muscle, Smooth/drug effects , Muscle, Smooth/metabolism , RNA, Messenger/genetics , Umbilical Arteries/drug effects , Vascular Capacitance/drug effects , Blotting, Western , Cells, Cultured , Calcium Channel Blockers/pharmacology , Calcium Channels/chemistry , Calcium Channels/genetics , Calcium Channels/metabolism , Calcium/metabolism , Histamine Agonists/pharmacology , Histamine/pharmacology , Membrane Proteins/genetics , Membrane Proteins/metabolism , Muscle, Smooth/cytology , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Sarcoplasmic Reticulum/drug effects , Sarcoplasmic Reticulum/metabolism , Serotonin Receptor Agonists/pharmacology , Serotonin/pharmacology , TRPC Cation Channels/genetics , TRPC Cation Channels/metabolism , Umbilical Arteries/cytology , Umbilical Arteries/metabolismABSTRACT
TRPV5 is believed to play an important role in the regulation of urinary calcium excretion. We assessed the effects of hydrochlorothiazide (HCTZ) on the expression of TRPV5, calbindin-D28K, and several sodium transporters in hypercalciuric rats. Sprague- Dawley rats were divided into 4 groups; control, HCTZ, high salt, and high salt with HCTZ group in experiment 1; control, HCTZ, high calcium (Ca), and high Ca with HCTZ group in experiment 2. To quantitate the expression of TRPV5, calbindin- D28K, and sodium transporters, western blotting was performed. In both experiments, HCTZ significantly decreased urinary calcium excretion. TRPV5 protein abundance decreased in all hypercalciuric rats, and restored by HCTZ in both high salt with HCTZ and high Ca with HCTZ group. Calbindin-D28K protein abundance increased in the high salt and high salt with HCTZ groups, but did not differ among groups in experiment 2. Protein abundance of NHE3 and NKCC2 decreased in all hypercalciuric rats, and were restored by HCTZ in only high Ca-induced hypercalciuric rats. In summary, protein abundance of TRPV5, NHE3, and NKCC2 decreased in all hypercalciuric rats. The hypocalciuric effect of HCTZ is associated with increased protein abundance of TRPV5 in high salt or calcium diet-induced hypercalciuric rats.
Subject(s)
Animals , Male , Rats , Biological Transport , Calcium/urine , Calcium Channels/chemistry , S100 Calcium Binding Protein G/biosynthesis , Hydrochlorothiazide/pharmacology , Hypercalciuria/therapy , Models, Biological , Rats, Sprague-Dawley , Sodium/metabolism , Sodium-Hydrogen Exchangers/chemistry , Sodium-Potassium-Chloride Symporters/metabolism , TRPV Cation Channels/biosynthesis , Thiazides/pharmacologyABSTRACT
Hypokalemic periodic paralysis (HOPP) is a rare disease characterized by reversible attacks of muscle weakness accompanied by episodic hypokalemia. Recent molecular work has revealed that the majority of familial HOPP is due to mutations in a skeletal muscle voltage-dependent calcium-channel: the dihydropyridine receptor. We report a 13-yr old boy with HOPP from a family in which 6 members are affected in three generations. Genetic examination identified a nucleotide 3705 C to G mutation in exon 30 of the calcium channel gene, CACNA1S. This mutation predicts a codon change from arginine to glycine at the amino acid position #1239 (R1239G). Among the three known mutations of the CACNA1S gene, the R1239G mutation was rarely reported. This boy and the other family members who did not respond to acetazolamide, showed a marked improvement of the paralytic symptoms after spironolactone treatment.
Subject(s)
Adolescent , Female , Humans , Male , Acetazolamide/pharmacology , Arginine/chemistry , Calcium Channels/chemistry , Codon , Exons , Family Health , Glycine/chemistry , Hypokalemia/metabolism , Hypokalemic Periodic Paralysis/diagnosis , Korea , Muscle, Skeletal/metabolism , Mutation , Pedigree , Protein Structure, Tertiary , Sequence Analysis, DNA , Spironolactone/pharmacologyABSTRACT
Effects of pH on vascular tone and L-type Ca2+ channels were investigated using Mulvany myograph and voltage-clamp technique in rabbit basilar arteries. In rabbitbasilar arteries, high K+ produced tonic contractions by 11+/-0.6 mN (mean+/-S.E., n=19). When extracellular pH (pHo) was changed from control 7.4 to 7.9 ([alkalosis]o), K+-induced contraction was increased to 128+/-2.1% of the control (n=13). However, K+-induced contraction was decreased to 73+/-1.3% of the control at pHo 6.8 ([acidosis]o, n=4). Histamine (10 micrometer) also produced tonic contraction by 11+/-0.6 mN (n=17), which was blocked by post-application of nicardipine (1 micrometer). [alkalosis]o and [acidosis]o increased or decreased histamine-induced contraction to 134+/-5.7% and 27+/-7.6% of the control (n=4, 6). Since high K+- and histamine-induced tonic contractions were affected by nicardipine and pHo, the effect of pHo on voltage-dependent L-type Ca2+ channel (VDCCL) was studied. VDCCL was modulated by pHo: the peak value of Ca2+ channel current (IBa) at a holding of 0 mV decreased in [acidosis]o by 41+/-8.8%, whereas that increased in [alkalosis]o by 35+/-2.1% (n=3). These results suggested that the external pH regulates vascular tone partly via the modulation of VDCC in rabbit basilar arteries.
Subject(s)
Animals , Rabbits , Arteries/pathology , Basilar Artery/pathology , Calcium/metabolism , Calcium Channels/chemistry , Electrophysiology , Histamine/chemistry , Hydrogen-Ion Concentration , Muscle Cells/cytology , Muscle Contraction , Muscle, Smooth/pathology , Patch-Clamp Techniques , Potassium/chemistry , Stress, Mechanical , Time FactorsABSTRACT
No abstract available.
Subject(s)
Humans , Mice , Animals , Calcium Channels/physiology , Calcium Channels/chemistry , Gene Expression Regulation , Ion Channel Gating/physiology , Mutation , Nervous System/metabolism , Nervous System Diseases/physiopathology , Nervous System Diseases/genetics , Neurons/physiologyABSTRACT
Los canales iónicos actúan acelerando el movimiento de los iones a través de las membranas biológicas. Cada canal iónico se activa por un estímulo específico (p. ej. eléctrico, mecánico, químico, etc.). Los receptores de membrana que actúan como canales iónicos (RMCI), pueden pasar a un estado denominado desensibilizado, cuando el agonista se encuentra ligado al receptor. El estado desensibilizado de un RMCI, como por ejemplo, el receptor nicotínico para la acetilcolina (nAChR), es un estado no funcional del canal y es un caso particular del denominado agotamiento de receptores "receptors rundown". La desensibilización de los RCMI sólo involucra una reducción de su actividad y no de su eliminación de la membrana. La desensibilización es importante en el control de la transmisión sináptica y en el desarrollo del sistema nervioso. En esta revisión se discuten los resultados más relevantes relacionados a su caracterización y modulación, de igual manera, algunos aspectos relacionados con los principales modelos cinéticos que le han tomado en consideración. Finalmente, se plantea la utilización de nuevas técnicas de biología molecular y electrofisiología para el estudio de la desensibilización y su importancia en los sistemas biológicos