Percutaneous angioplasty and stenting of subclavian arteries before surgical coronary revascularization in a patient with an aberrant right subclavian artery.

An aberrant right subclavian artery occurs in less than 2% of the population. An associated stenosis of the subclavian artery carries a risk of subclavian-coronary steal in patients who undergo coronary revascularization. We report on the case of a 54-year-old man admitted to our hospital for a coronary artery bypass graft (CABG). Angiographic examination revealed bilateral subclavian-artery stenosis with an aberrant right subclavian artery, anomalous origin of the right vertebral artery from the right common carotid artery, and left vertebral-artery occlusion. The patient underwent successful bilateral subclavian angioplasty and stenting.

Inhibitory effect of nitrovasodilators and cyclic GMP on ET-1-activated Ca(2+)-permeable nonselective cation channel in rat aortic smooth muscle cells.

1. In single vascular smooth muscle cells (VSMCs) isolated from the aortae of male Wistar rats, we examined the effects of nitric oxide (NO) donors such as sodium nitroprusside (SNP) and S-nitroso-N-acetyl-DL-penicillamine (SNAP), and 8-bromo-guanosine-3':5'-cyclic monophosphate (8-bromo-cyclic GMP) on endothelin-1 (ET-1)-activated Ca(2+)-permeable nonselective cation channel by use of whole-cell recordings of patch-clamp technique and monitoring of intracellular free Ca(2+)-concentration ([Ca2+]i) with fura-2 real-time digital microfluorometry. 2. ET-1 evoked an initial transient peak and a subsequent sustained elevation in [Ca2+]i. After removal of extracellular Ca2+. ET-1 evoked only an initial transient peak without a sustained phase. Nifedipine (1 microM), a specific blocker of the L-type voltage-operated Ca2+ channel (VOC), reduced the sustained phase to about 40% of the control level. The remaining part of the sustained phase was abolished by 30 microM SK&F 96365, a blocker of nonselective cation channels. 3. The nifedipine-resistant sustained elevation in [Ca2+]i was abolished by 100 microM SNP, 10 microM SNAP and 300 microM 8-bromo-cyclic GMP. Neither SNP, SNAP nor 8-bromo-cyclic GMP significantly affected the basal level of [Ca2+]i. 4. In a VSMC clamped at a holding potential of -60 mV with K+ in the pipette solution replaced by Cs+, application of 10(-8) M ET-1 induced an inward current with an increase in baseline fluctuation. With fluctuation analysis, unit conductance of the ET-1-induced current was calculated to be about 21 pS. The ET-1-induced current was linearly related to the membrane potentials with its reversal potential of -5.5 mV. 5. The ET-1-induced current was reversibly and completely inhibited by 30 microM SK&F 96365 or 500 microM Cd2+. The current inhibited by SK&F 96365 or Cd2+ was linearly related to membrane potential with a reversal potential of about -5 mV. 6. The ET-1-induced current was reversibly and completely inhibited by 100 microM SNP, 10 microM SNAP and 300 microM 8-bromo-cyclic GMP. The current inhibited by SNP, SNAP or 8-bromo-cyclic GMP showed linear voltage-dependence and reversed at about -5 mV. 7. In a bath solution in which all cations were replaced by 30 mM Ca2+ and 100 mM nonpermeant cation N-methyl-D-glucamine (NMDG), ET-1 evoked a current with a reversal potential of -11 mV, from which PCa2+/Pcs1 was calculated to be 2.1. This Ca2+ current was also abolished by 100 microM SNP, 10 microM SNAP and 300 microM 8-bromo-cyclic GMP. The current inhibited by SNP, SNAP or 8-bromo-cyclic GMP showed linear voltage-dependence and reversed at about -11 mV. 8. These results taken together indicate that NO through a cyclic GMP signalling pathway inhibits ET-1-activated Ca(2+)-permeable nonselective cation channels, thereby suppressing the sustained increase in [Ca2+]i. Thus, the present study indicates that this Ca(2+)-permeable nonselective cation channel is an important target for nitrovasodilators.

Pharmacological characterization of Ca2+ entry channels in endothelin-1-induced contraction of rat aorta using LOE 908 and SK&F 96365.

We have recently shown that endothelin-1 (ET-1) activates two types of Ca2+-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and store-operated Ca2+ channel (SOCC). These channels can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. Here we characterized Ca2+ entry channels involved in ET-1-induced contractions of rat thoracic aortic rings and increases in the intracellular free Ca2+ concentration ([Ca2+]i) of single smooth muscle cells using these blockers. LOE 908 or a blocker of voltage-operated Ca2+ channel nifedipine had no effect on the contractions and increases in [Ca2+]i induced by thapsigargin or ionomycin, whereas SK&F 96365 abolished them. The contractions and increases in [Ca2+]i induced by ET-1 depended on extracellular Ca2+ but were resistant to nifedipine. The responses to lower concentrations (< or =0.1 nM) of ET-1 were abolished by either SK&F 96365 or LOE 908. The responses to higher concentrations (> or = 1 nM) were abolished by SK&F 96365, but were partially resistant to LOE 908. SK&F 96365 inhibited the LOE 908-resistant contractions induced by higher concentrations of ET-1 with IC50 values similar to those for contractions induced by thapsigargin or ionomycin. These results show that the contractions and increases in [Ca2+]i of rat aortic smooth muscles at lower concentrations of ET-1 involve only one Ca2+ entry channel which is sensitive to SK&F 96365 and LOE 908 (NSCC-2), whereas those at higher concentrations of ET-1 involve another Ca2+ entry channel which is sensitive to SK&F 96365 but resistant to LOE 908 (SOCC) in addition to the former channel.

Activation of three types of voltage-independent Ca2+ channel in A7r5 cells by endothelin-1 as revealed by a novel Ca2+ channel blocker LOE 908.

1. We have shown that in addition to voltage-operated Ca2+ channel (VOC), endothelin-1 (ET-1) activates two types of Ca2+-permeable nonselective cation channel (NSCC) in A7r5 cells: its lower concentrations (< or = 1 nM; lower [ET-1]) activate only an SK&F 96365-resistant channel (NSCC-1), whereas its higher concentrations (> or = 10 nM; higher [ET-1]) activate an SK&F 96365-sensitive channel (NSCC-2) as well. 2. We now characterized the effects of a blocker of Ca2+ entry channel LOE 908 on NSCCs and store-operated Ca2+ channel (SOCC) in A7r5 cells, and using two drugs, clarified the involvement of these channels in the ET-1-induced increase in the intracellular free Ca2+ concentrations ([Ca2+]i). Whole-cell recordings and [Ca2+]i monitoring with fluo-3 were used. 3. LOE 908 up to 10 microM had no effect on increases in [Ca2+]i induced by thapsigargin or ionomycin, but SK&F 96365 abolished them. 4. In the cells clamped at -60 mV, both lower and higher [ET-1] induced inward currents with linear iv relationships and the reversal potentials of -15.0 mV. Thapsigargin induced no currents. 5. In the presence of nifedipine, lower [ET-1] induced a sustained increase in [Ca2+]i, whereas higher [ET-1] induced a transient peak and a sustained increase. The sustained increases by lower and higher [ET-1] were abolished by removal of extracellular Ca2+, and they were suppressed by LOE 908 to 0 and 35%, respectively, with the LOE 908-resistant part being abolished by SK&F 96365. 6. These results show that LOE 908 is a blocker of NSCCs without effect on SOCC, and that the increase in [Ca2+]i at lower [ET-1] results from Ca2+ entry through NSCC-1 in addition to VOC, whereas the increase at higher [ET-1] involves NSCC-1, NSCC-2 and SOCC in addition to VOC.

Activation of two types of Ca2+-permeable nonselective cation channel by endothelin-1 in A7r5 cells.

1. In A7r5 cells loaded with the Ca2+ indicator fura-2, we examined the effect of a Ca2+ channel blocker SK&F 96365 on increases in intracellular free Ca2+ concentrations ([Ca2+]i) and Mn2+ quenching of fura-2 fluorescence by endothelin-1 (ET-1). Whole-cell patch-clamp was also performed. 2. Higher concentrations (> or = 10 nM) of ET-1 (higher [ET-1]) evoked a transient peak and a subsequent sustained elevation in [Ca2+]i: removal of extracellular Ca2+ abolished only the latter. A blocker of L-type voltage-operated Ca2+ channel (VOC) nifedipine at 1 microM reduced the sustained phase to about 50%, which was partially sensitive to SK&F 96365 (30 microM). 3. Lower [ET-1] (< or = 1 nM) evoked only a sustained elevation in [Ca2+]i which depends on extracellular Ca2+. The elevation was partly sensitive to nifedipine but not SK&F 96365. 4. In the presence of 1 microM nifedipine, higher [ET-1] increased the rate of Mn2+ quenching but lower [ET-1] had little effect. 5. In whole-cell recordings, both lower and higher [ET-1] induced inward currents at a holding potential of -60 mV with linear I-V relationships and reversal potentials close to 0 mV. The current at lower [ET-1] was resistant to SK&F 96365 but was abolished by replacement of Ca2+ in the bath solution with Mn2+. The current at higher [ET-1] was abolished by the replacement plus SK&F 96365. 6. In a bath solution containing only Ca2+ as a movable cation, ET-1 evoked currents: the current at lower [ET-1] was sensitive to Mn2+, whereas that at higher [ET-1] was partly sensitive to SK&F 96365. 7. These results indicate that in addition to VOC, ET-1 activates two types of Ca2+-permeable nonselective cation channel depending on its concentrations which differ in terms of sensitivity to SK&F 96365 and permeability to Mn2+.

Role of nonselective cation channels as Ca2+ entry pathway in endothelin-1-induced contraction and their suppression by nitric oxide.

The present study was carried out to clarify the role of nonselective cation channels as a Ca2+ entry pathway in the contraction and the increase in [Ca2+]i induced by endothelin- in endothelium-denuded rat thoracic aorta rings, and their suppression by nitric oxide (NO). In Ca2+-free medium, the endothelin-1-induced contraction was suppressed to about 20% of control values, although the increase in [Ca2+]i became negligible. The contraction and the increase in [Ca2+]i monitored by fura 2 fluorescence were unaffected by a blocker of L-type voltage-operated Ca2+ channels nifedipine. A blocker of nonselective cation channels 1-[beta-[3-(4-methoxyphenyl)propoxyl]-4-methoxyphenethyl]-1H-imida zole . HCl(SK&F 96365) suppressed the endothelin-1-induced contraction and increase in [Ca2+]i to the level similar to that after removal of extracellular Ca2+. SK&F 96365 had no further effect on the endothelin-1-induced contraction in the absence of extracellular Ca2+. The endothelin-1-induced contraction and increase in [Ca2+]i were abolished by a donor of NO sodium nitroprusside. The effects of another NO donor 3-morpholinosydnonimine (SIN-1) were also tested and yielded essentially similar results to those for sodium nitroprusside on the endothelin-1-induced contraction. Furthermore, the inhibitory effects of sodium nitroprusside could be blocked with a guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ) at 30 microM. These findings suggest that Ca2+ entry through nonselective cation channels but not voltage-operated Ca2+ channels plays a critical role in the endothelin-1-induced increase in [Ca2+]i and the resulting contraction and that inhibition by NO of the endothelin-1-induced contraction is mainly the result of blockade of Ca2+ entry through these channels.

Failure of endothelin-1 to activate store-operated Ca2+ channels by lack of mobilization from intracellular Ca2+ stores in cultured bovine adrenal chromaffin cells.

We have recently shown that in addition to L-type voltage-operated Ca2+ channel (VOC), endothelin-1 (ET-1) stimulation opens two types of Ca2+-permeable nonselective cation channels [designated nonselective cation channel-1 (NSCC-1) and NSCC-2]. However, in this Ca2+ entry, the involvement of store-operated Ca2+ channel (SOCC), which is suggested to exist in chromaffin cells, was unclear. Those NSCCs as well as SOCC can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. To clarify whether SOCC should actually exist and play a role in Ca2+ entry in chromaffin cells stimulated with ET-1, we examined the effects of removal of extracellular Ca2+, thapsigargin (TG, an inhibitor of endoplasmic reticulum Ca2+-ATPase), LOE 908 and SK&F 96365 on cytosolic free Ca2+ concentrations ([Ca2+]i) in cultured bovine adrenal chromaffin cells. After the cells were exposed to Ca2+-free medium followed by exposure to TG to deplete Ca2+ from the intracellular Ca2+ store, restoration of extracellular Ca2+ caused a gradual increase in [Ca2+]i (to about 200% of control). The increase was unaffected by LOE 908, but completely abolished by SK&F 96365. In the Ca2+-free medium, no increase in [Ca2+]i by ET-1 was observed, but the subsequent restoration of extracellular Ca2+ induced a rapid increase in [Ca2+]i (to the same level of [Ca2+]i as that evoked by ET-1 in the normal medium (1.0 mM Ca2+)). Since SK&F 96365 is also a blocker of SOCC, these results indicate that in bovine adrenal chromaffin cells, Ca2+ entry through SOCC (Ca2+ influx through the capacitative Ca2+ entry system) occurs but is comparably weak, and that it virtually does not work on the stimulation of ET-1.

Pharmacological characterization of receptor-mediated Ca2+ entry in endothelin-1-induced catecholamine release from cultured bovine adrenal chromaffin cells.

To clarify the mechanism for the endothelin-1 (ET-1)-induced release of catecholamines from the adrenal gland, we examined the effects of removal of extracellular Ca2+, blockers of L-, N-, P- and Q-types of voltage-operated Ca2+ channels (VOCC) such as nifedipine (L-type), omega-conotoxin GVIA (N-type), omega-agatoxin IVA (P-type) and omega-conotoxin MVIIC (Q-type) and blockers of voltage-independent Ca2+ entry channel such as SK&F 96365 and LOE 908 on release of catecholamines, the cytosolic free Ca2+ concentration ([Ca2+]i), and 45Ca2+ uptake in cultured bovine adrenal chromaffin cells. ET-1 but not ET-3 induced increases in release of catecholamines, [Ca2+]i, and 45Ca2+ uptake. The responses to ET-1 were abolished by the antagonist for ET(A) receptors, BQ-123, but not by the antagonist for ET(B) receptors, BQ-788, and they were abolished by removal of extracellular Ca2+. The increases were only partially inhibited (to about 65% of control) by nifedipine but unaffected by any of the omega-toxins. The nifedipine-resistant increase was inhibited by SK&F 96365 (to about 40%) and abolished by LOE 908 alone. These results indicate that ET-1 augments the release of catecholamines from adrenal chromaffin cells through ET(A) receptors, by activating two types of Ca2+ entry channels in addition to L-type VOCC: one (nonselective cation channel-1; NSCC-1) is sensitive to LOE 908 but resistant to SK&F 96365, whereas the other (NSCC-2) is sensitive to both LOE 908 and SK&F 96365.

Transient ischemic attack due to dissection of the middle cerebral artery--case report.

A 57-year-old man presented with a transient ischemic attack due to dissection of the middle cerebral artery. He suffered total aphasia and clouding of consciousness for several minutes. On admission, he was alert without neurological deficit. Magnetic resonance (MR) angiography and conventional angiography depicted irregularity and double lumen of the left middle cerebral artery. The diagnosis was dissection of the middle cerebral artery. After 1 month, he left our institute with no neurological deficit. Transient ischemic attack associated with dissection of an intracranial artery is unusual. The source images of MR angiography are useful for the essential follow up of dissection.

[Recurrence of a giant fusiform aneurysm after neck clipping: case report].

The patient was a 71-year-old female. On December 20, 1995, she suddenly developed a severe headache with vomiting and was transferred to our hospital. On admission, her conciousness level was 1-2 on the Japan Coma Scale, but there was no neurological deficit except for right oculomotor palsy. Computed tomography showed subarachnoid hemorrhage which had permeated the right lateral ventricle. On cerebral angiography, a giant fusiform aneurysm in the right internal carotid artery was recognized. During the emergency operation, neither neck clipping nor carotid reconstruction was possible because of the tight adhesion of the aneurysm to the peripheral tissue. On account of this, proximal clipping of the carotid artery with external carotid-middle cerebral artery anastomosis with saphenous vein graft was selected. This patient had had an episode of subarachnoid hemorrhage owing to rupture of the right internal carotid-posterior communicating artery aneurysm ten years earlier. At that time, the aneurysmal neck was clipped with a slight residual neck and she left the hospital on foot. Five days later, when the aneurysm was found to be completely thrombosed on CT scan, antiplatelet therapy was started. Although low density areas which corresponded to the regions fed by the right anterior choroidal artery were presented, re-rupture did not occur. Follow-up angiography showed that the aneurysm was completely thrombosed and that the right middle cerebral and the anterior cerebral artery blood was circulated via the vein graft. Among recurrent cases of aneurysm after neck clipping, it is unusual for a giant fusiform aneurysm to be recognized. The growth may have been caused by sclerotic change of the arterial wall. Oculomotor palsy may have delayed the detection of the recurrence of the aneurysm. When residual neck is presented on follow-up angiography, the next angiography should be carried out within at least three years. In this case, antiplatelet therapy was effective to prevent thromboembolism from the aneurysm.

[Vertebral artery dissection without subarachnoid hemorrhage studied by serial angiography].

We presented six cases we have encountered of vertebral artery dissection without subarachnoid hemorrhage followed by serial angiography. All six patients suffered from severe headache and/or nuchal pain at onset. Pain was acute at onset, with severe intensity and sharp quality and was located unilaterally on the dissection side. Only two patients showed neurological deficits. CT scan disclosed no abnormalities in any of the cases and angiography performed at the acute stage showed varied findings. The so-called pearl and string sign, which has been considered the most common finding of this disorder, was observed in only two cases. The definite diagnosis was able to be finally made by serial angiography. During the first few months after onset, dynamic changes of angiographical findings were demonstrated in all cases. During these periods, although transient deterioration of the angiographical findings was shown in four cases, spontaneous healing or improvement was finally recognized in all cases except one which progressed to total occlusion. In contrast to this, in the chronic stage, no changes of angiographical findings occurred in any of the cases. All six patients were treated non-surgically. All showed almost total recovery from symptoms and returned to their previous life styles. There were no cases of recurrent deterioration of symptoms in spite of the changes indicated by the angiographical findings. Our experience with these patients suggests that the actual incidence of vertebral artery dissection without subarachnoid hemorrhage may be much higher than is usually thought, and that the natural course of this disorder seems to be usually favorable. However, longer follow-up study and an analysis of a larger number of cases are required to identify the true incidence and the natural course of this disorder. In addition, we would like to emphasize that accurate diagnosis is most important. If it is clinically suspected, careful investigations including serial angiography should be carried out for the diagnosis of this disorder.

[Infantile brainstem infarction due to vertebral artery fenestration: a case report].

The patient was a 10-year-old boy. He had a sudden onset of generalized tonic-clonic convulsion. On admission, he presented with mild disturbance of consciousness and right hemiparesis. The electroencephalogram showed diffuse spike and slow wave complex. Three days later, magnetic resonance imaging (MRI) showed high signal intensity areas in the left cerebellar hemisphere and pons. On cerebral angiograms, coiling of the bilateral carotid arteries, occlusion of the basilar artery at the distal site, and fenestration of the left vertebral artery at the atlanto-axis level were shown. According to these findings, the patient was diagnosed with brainstem and cerebellar infarction. On the follow-up angiograms, recanalization of the basilar artery on the right vertebral angiogram, occlusion of the left vertebral artery at the distal site of the posterior inferior cerebellar artery origin, and occlusion of one of the duplicated vertebral arteries were recognized. Fenestration of the vertebral artery is presented in about 1-2% at angiography and autopsy and the clinical significance is controversial. In this case, the fenestration may have played a role as an embolic source because there was no probable cause of the cerebral infarction, and the vascular occlusion and recanalization occurred near the distal site of the fenestration. These findings suggest the clinical significance of vertebral artery fenestration as an embolic source.

Near-infrared spectroscopy in carotid artery stenting predicts cerebral hyperperfusion syndrome.

OBJECTIVE: Cerebral hyperperfusion syndrome (CHS) following carotid artery stenting (CAS) or carotid endarterectomy (CEA) is rare but often fatal once intracranial hemorrhage has occurred. In particular, CHS occurs significantly earlier after CAS than after CEA. Thus a monitoring method for early detection of CHS is required. Near-infrared spectroscopy (NIRS) provides a noninvasive monitoring technique for assessing regional cerebral oxygen saturation (rSO2). This study evaluated the usefulness of transcranial NIRS during CAS for prediction of CHS. METHODS: Periprocedural rSO2 was monitored in 64 cases of CAS (52 men, 12 women; 71 +/- 6.6 years). The average degree of carotid stenosis was 76.8 +/- 11.3% by North American Symptomatic Carotid Endarterectomy Trial criteria. Bifrontal rSO2 was monitored during the procedure using NIRS. Seventeen patients were symptomatic and 47 were asymptomatic. CHS was diagnosed by increased cerebral blood flow by SPECT performed on the day after treatment with deterioration of neurologic symptoms. RESULTS: CHS was observed in two cases (3.1%). In the CHS group, post-reperfusion rSO2 values increased >24% from baseline until 3 minutes after reperfusion. In the non-CHS group, the normal upper limit (NUL) of the rSO2 change was set at 10.0% at 3 minutes after reperfusion. In the CHS group, rSO2 at 3 minutes after reperfusion was markedly higher than the NUL. In patients showing an rSO2 at 3 minutes after reperfusion increased by more than 10.0%, CHS following CAS could be predicted. CONCLUSION: Periprocedural increases in regional cerebral oxygen saturation measured by near- infrared spectroscopy can be an excellent predictor of cerebral hyperperfusion syndrome after carotid artery stenting.

Result of neck clipping and coil embolization as a treatment for unruptured aneurysm.

SUMMARY: The report of the International Subarachnoid Aneurysm Trial (ISAT) study showed that coil embolization was superior to neck clipping as a treatment for subarachnoid hemorrhage (SAH) (1). Recently, some results of treatments for unruptured aneurysm via coil embolization and neck clipping have been reported (2,3).We compared the results of coil embolization and neck clipping in our institute. Generally better outcomes were obtained by endovascular surgery than by neck clipping. Postoperative ischemic strokes occurred in one case (1.8%) as a major stroke and in three cases (5.6% ) as a minor stroke among coil-treated cases, and in two cases(2.6%) as a major stroke, and in seven cases(9.0%) as a minor stroke among neck clipping cases. Other complications after these treatments were six cases of subdural effusion/hematoma, four cases of infection, two cases of epidural hematoma, one abducens nerve palsy, one hydrocephalus, and one acute myocardial infarction among 78 neck clipping cases, and two subcutaneous hematoma, one pseudoaneurysm at the puncture points, one direct carotid-cavernous fistula among 54 coil-treated cases. Four coil-treated cases, in which introduction of microcatheters to the aneurysm was impossible, were treated completely by neck clipping after endovascular treatments. In terms of modified Rankin Scale(mRS) three months after treatments, while mRS 3 was noted in only one case in the endovascular treatment group, there were one case of mRS 3, two cases of mRS 4, and two cases of mRS 5 in the neck clipping group. Duration of hospitalization averaged 11.9 days in the endovascular group and 24.1 days in the neck clipping group. The results of endosaccular enbolizations as treatment of the unruptured aneurysm seems to be better than neck clipping. However, not all cases of unruptured aneurysms can be treated by coil embolization due to the width of aneurysmal neck and relation of the aneurysm to parent arteries. Therefore, surgeons should also be able to perform neck clipping as an alternative modality.

Result of neck clipping and coil embolization as a treatment for subarachnoid haemorrhage.

SUMMARY: The report of the International Subarachnoid Aneurysm Trial (ISAT) study showed that coil embolization was superior to neck clipping as a treatment for subarachnoid haemorrhage (SAH) (1). We compared the results of coil embolization and neck clipping in our institute. Generally better outcomes were obtained by endovascular surgery than neck clipping. Symptomatic vasospasm and symptomatic hydrocephalus occurred less frequently in coil embolization than neck clipping. Because not all cases of SAH can be treated by coil embolization due to the width of aneurysmal neck and relation of the aneurysm to parent arteries, we should also be able to perform neck clipping as another modality.

Endothelin-1-induced contraction of rat thoracic aorta depends on calcium entry through three types of calcium channel.

We have recently shown that endothelin-1 (ET-1) activates two types of Ca2+-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and store-operated Ca2+ channel (SOCC). These channels can be pharmacologically discriminated using 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1-H-imidazoe l hydrochloride (SK&F 96365) (a blocker of NSCC-2 and SOCC) and (RS)-(3,4-dihydro-6,7-dimethoxyisoquinoline-1-gamma l)-2-phenyl-N,N-di-[2-(2,3,4-trimethoxyphenyl)ethyl]acetamide (LOE 908) (a blocker of NSCC-1 and NSCC-2). For our study we characterized Ca2+ channels involved in ET-1-induced contractions and increases in the intracellular free Ca2+ concentration ([Ca2+]i) using these blockers. Our results show that the response to lower concentrations of ET-1 involves only one Ca2+ channel which is sensitive to SK&F 96365 and LOE 908 (NSCC-2). In contrast, the response to higher concentrations of ET-1 involves two types of Ca2+ channel in addition to NSCC-2: one is sensitive to SK&F 96365 but resistant to LOE 908 (SOCC), and the other is resistant to SK&F 96365 but sensitive to LOE 908 (NSCC-1). Furthermore, the percentage contribution of Ca2+ entry through NSCC-1, NSCC-2 and SOCC is calculated to be 10%, 50-60% and 30-40%, respectively.

Pharmacological properties of calcium entry channels in A7r5 cells activated by endothelin-1.

Using whole-cell recordings of patch-clamp and monitoring of the intracellular free calcium (Ca2+) concentration ([Ca2+]i), we characterized Ca2+ entry channels in A7r5 cells activated by endothelin-1 (ET-1). ET-1 activates three types of voltage-independent Ca2+ entry channels: two types of Ca2+-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and store-operated Ca2+ channel (SOCC). Furthermore, it was found that these channels can be discriminated pharmacologically using Ca2+ channel blockers such as 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl)-1-H-imidazoe l hydrochloride (SK&F 96365) and (RS)-(3,4-dihydro-6,7-dimethoxyisoquinoline-1-gamma l)-2-phenyl-N,N-di-[2-(2,3,4-trimethoxyphenyl)ethoxyl]acetamide (LOE 908). NSCC-1 is resistant to SK&F 96365 but sensitive to LOE 908, whereas NSCC-2 is sensitive to both drugs: SOCC is sensitive to SK&F 96365 but resistant to LOE 908. Using these channel blockers, we analyzed the Ca2+ entry channels involved in the ET-1-induced increase in [Ca2+]i of the cells. The increase induced by lower concentrations of ET-1 (< or = 0.1 nM) was unaffected by SK&F 96365 but it was abolished by LOE 908. In contrast, the increase caused by higher concentrations of ET-1 (> or = 1 nM) was suppressed by SK&F 96365 or LOE 908 to about 35% of controls, and abolished by combined treatment with SK&F 96365 and LOE 908. These results show that the increase in [Ca2+]i resulting from lower concentrations of ET-1 (< or = 0.1 nM) involves Ca2+ entry through only NSCC-1, whereas that resulting from higher concentrations of ET-1 involves Ca2+ entry through NSCC-1, NSCC-2 and SOCC, contributing 35%, 30% and 30%, respectively, to total Ca2+ entry.

[Characterization of voltage-independent Ca2+ channels activated by endothelin-1].

To clarify Ca2+ entry channels involved in the endothelin-1 (ET-1)-induced increase in the intracellular concentration ([Ca2+]i), we performed whole-cell recordings of patch-clamp techniques and monitoring of [Ca2+]i with Ca2+ indicators fura-2 and fluo-3 in A7r5 cells (a cell line derived from rat thoracic aortic smooth muscle cells). With whole-cell recordings, lower concentrations (< or = 1 nM) of ET-1 activated a Ca(2+)-permeable nonselective cation channel (designated NSCC-1). In contrast, higher concentrations (> or = 1 nM) of ET-1 activated two types of Ca(2+)-permeable nonselective cation channel (designated NSCC-1 and NSCC-2) and store-operated Ca2+ channel (SOCC). Importantly, we found that these Ca2+ channels can be pharmacologically discriminated using blockers of the so-called receptor operated Ca2+ influx such as SK&F 96365 and LOE 908. That is, NSCC-1 is resistant to SK&F 96365 but sensitive to LOE 908; NSCC-2 is sensitive to both SK&F 96365 and LOE 908; SOCC is sensitive to SK&F 96365 but resistant to LOE 908. Using these blockers, we analyzed the ET-1-induced increase in [Ca2+]i. The increase in [Ca2+]i induced by lower concentrations of ET-1 was resistant to SK&F 96365 but sensitive to LOE 908. In contrast, the increase in [Ca2+]i induced by higher concentrations of ET-1 was partially suppressed to approximately 30% of controls by either SK&F 96365 or LOE 908 alone, and it was abolished by their combination. These results show that the increase in [Ca2+]i induced by lower concentrations (< or = 1 nM) of ET-1 results from Ca2+ influx through NSCC-1, whereas the increase in [Ca2+]i induced by higher concentrations (> or = 10 nM) of ET-1 results from Ca2+ influx through NSCC-1, NSCC-2 and SOCC.

[Analysis of Ca2+ entry channels in endothelin-1-induced contraction of rat aorta using new blockers].

1) We have recently shown that endothelin-1 (ET-1) activates two types of Ca(2+)-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and store-operated Ca2+ channel (SOCC). These channels can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. Here we characterized Ca2+ entry channels involved in ET-1-induced contractions of rat thoracic aortic rings and increases in the intracellular free Ca2+ concentration ([Ca2+]i) of single smooth muscle cells using these blockers. 2) LOE 908 or a blocker of voltage-operated Ca2+ channel nifedipine had no effect on the contractions and increases in [Ca2+]i induced by thapsigargin, whereas SK&F 96365 abolished them. 3) The contractions and increases in [Ca2+]i induced by ET-1 depended on extracellular Ca2+ but were resistant to nifedipine. The responses to lower concentrations (< or = 0.1 nM) of ET-1 were abolished by either SK&F 96365 or LOE 908. The responses to higher concentrations (> or = 1 nM) were abolished by SK&F 96365, but were partially resistant to LOE 908. 4) These results show that the contractions and increases in [Ca2+]i of rat aortic smooth muscles at lower concentrations of ET-1 involve only one Ca2+ entry channel which is sensitive to SK&F 96365 and LOE 908 (NSCC-2), whereas those at higher concentrations of ET-1 involve another Ca2+ entry channel which is sensitive to SK&F 96365 but resistant to LOE 908 (SOCC) in addition to the former channel.

Ca2+ entry channels in rat thoracic aortic smooth muscle cells activated by endothelin-1.

The contraction of the rat aorta induced by endothelin-1 (ET-1) requires entry of extracellular Ca2+, but involvement of voltage-operated Ca2+ channel is minor. Using whole-cell recordings of patch-clamp and monitoring of the intracellular free Ca2+ concentration ([Ca2+]i), we characterized Ca2+ entry channels in A7r5 cells activated by ET-1. ET-1 activates three types of voltage-independent Ca2+ entry channels: two types of Ca2+-permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and a store-operated Ca2+ channel (SOCC). Furthermore, it was found that these channels can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. NSCC-1 is resistant to SK&F 96365, but sensitive to LOE 908, whereas NSCC-2 is sensitive to both SK&F 96365 and LOE 908. SOCC is sensitive to SK&F 96365, but resistant to LOE 908. Using these channel blockers, we analyzed Ca2+ entry channels involved in the ET-1-induced contractions of rat thoracic aorta and increases in [Ca2+]i of single smooth muscle cells. The responses to lower concentrations of ET-1 (< or = 0.1 nM) were abolished by either SK&F 96365 or LOE 908 alone. In contrast, the responses to higher concentrations of ET-1 (> or = 1 nM) were suppressed by SK&F 96365 or LOE 908 to about 10% and 35% of controls, respectively, and abolished by combined treatment with SK&F 96365 and LOE 908. These results show that the responses of rat aorta to lower concentrations of ET-1 involve only one Ca2+ channel that is sensitive to SK&F 96365 and LOE 908 (NSCC-2), whereas those to higher concentrations of ET-1 involve NSCC-1, NSCC-2 and SOCC, contributing 10%, 55% and 35%, respectively, to total Ca2+ entry.