Agonist-activated, ryanodine-sensitive, IP3-insensitive Ca2+ release channels in longitudinal muscle of intestine

1994 ◽  
Vol 266 (5) ◽  
pp. C1421-C1431 ◽  
Author(s):  
J. F. Kuemmerle ◽  
K. S. Murthy ◽  
G. M. Makhlouf
Keyword(s):  
Longitudinal Muscle ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Cell Types ◽  
Ruthenium Red ◽  
Effective Concentration ◽  
Channel Blockers ◽  
Inositol 1,4,5 Trisphosphate ◽  
Dissociation Constant Kd ◽  
Ca2 Channels

We have previously shown that Ca2+ mobilization in longitudinal muscle is not mediated by inositol 1,4,5-trisphosphate (IP3) and depends on an obligatory influx of Ca2+. The present study examined whether Ca2+ influx activates ryanodine-sensitive Ca2+ channels to cause Ca(2+)-induced Ca2+ release. Ryanodine bound with high affinity to longitudinal muscle cells [dissociation constant (Kd) 7.3 +/- 0.3 nM] and microsomes (Kd 7.5 +/- 0.4 nM) and induced concentration-dependent 45Ca2+ efflux [50% effective concentration (EC50) 1.3 +/- 0.5 nM], increase in cytosolic free Ca2+ (EC50 2.0 +/- 0.7 nM), and contraction (EC50 0.9 +/- 0.2 nM) but had no effect in circular muscle cells. Ryanodine binding and ryanodine-induced Ca2+ release were enhanced by caffeine and inhibited by dantrolene and ruthenium red but were not affected by IP3 or heparin. Changes in Ca2+ concentration (50-500 nM) caused Ca2+ release from permeabilized longitudinal but not circular muscle cells loaded with 45Ca2+. The contractile agonist cholecystokinin-8 elicited 45Ca2+ efflux in both circular and longitudinal muscle cells; efflux in longitudinal muscle cells was abolished by Ca2+ channel blockers and by pretreatment of the cells with ryanodine. Pretreatment with thapsigargin abolished agonist-induced 45Ca2+ efflux in both cell types. We conclude that ryanodine-sensitive IP3-insensitive Ca2+ release channels with properties similar to those in cardiac muscle are present in longitudinal but not circular muscle cells of intestine and that agonist-mediated Ca2+ influx activates these channels, leading to Ca(2+)-induced Ca2+ release.

Agonist-mediated activation of PLA2 initiates Ca2+ mobilization in intestinal longitudinal smooth muscle

1995 ◽  
Vol 269 (1) ◽  
pp. G93-G102 ◽  
Author(s):  
K. S. Murthy ◽  
J. F. Kuemmerle ◽  
G. M. Makhlouf
Keyword(s):  
Smooth Muscle ◽  
Longitudinal Muscle ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Cell Types ◽  
Initial Increase ◽  
Channel Blockers ◽  
Longitudinal Smooth Muscle ◽  
Highly Sensitive ◽  
Cyclic Adp Ribose

Recent studies have shown that Ca2+ mobilization in longitudinal muscle is initiated by inositol 1,4,5-trisphosphate (IP3)-independent Ca2+ influx that acts as a trigger for Ca(2+)-induced Ca2R release. The present study examined whether arachidonic acid (AA) acts as mediator of the initial Ca2+ influx. Cholecystokinin octapeptide caused transient concentration-dependent increase in AA release in dispersed intestinal longitudinal but not circular muscle cells followed by sustained increase in both muscle cell types. The initial increase in AA release coincided with the initial Ca2+ transient and muscle contraction: all three events were abolished by guanosine 5'-O-(2-thiodiphosphate), pertussis toxin (PTX), and the phospholipase A2 (PLA2) inhibitor, dimethyleicosadienoic acid, but were not affected by calphostin C or neomycin. Exogenous AA caused concentration-dependent contraction and increase in cytosolic free Ca2+ ([Ca2+]i) in longitudinal but not circular muscle cells; both events were abolished by Ca2+ channel blockers. Depletion of Ca2+ stores with thapsigargin attenuated with thapsigargin attenuated agonist- and AA-mediated increase in [Ca2+]i and contraction in longitudinal muscle cells: the residual [Ca2+]i increase (35%) and contraction (25%) reflected the component of Ca2+ influx. We conclude that AA released by agonist-mediated G protein-dependent PTX-sensitive activation of PLA2 mediates Ca2+ influx, which then triggers Ca(2+)-induced Ca2+ release. The process is independent of phosphatidylinositol hydrolysis and occurs exclusively in longitudinal smooth muscle, in which Ca2+ release channels are highly sensitive to Ca2+, ryanodine, and cyclic ADP-ribose and insensitive to IP3.

Impairment of Ca2+ mobilization in circular muscle cells of the inflamed colon

2000 ◽  
Vol 278 (2) ◽  
pp. G234-G242 ◽  
Author(s):  
Xuan-Zheng Shi ◽  
Sushil K. Sarna
Keyword(s):  
Contractile Response ◽  
Single Cells ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Ruthenium Red ◽  
Cell Length ◽  
Muscle Contractility ◽  
Inositol 1,4,5 Trisphosphate ◽  
Intracellular Ca

This study investigated whether inflammation modulates the mobilization of Ca2+ in canine colonic circular muscle cells. The contractile response of single cells from the inflamed colon was significantly suppressed in response to ACh, KCl, and BAY K8644. Methoxyverapamil and reduction in extracellular Ca2+concentration dose-dependently blocked the response in both normal and inflamed cells. The increase in intracellular Ca2+concentration in response to ACh and KCl was significantly reduced in the inflamed cells. However, Ca2+ efflux from the ryanodine- and inositol 1,4,5-trisphosphate (IP3)-sensitive stores, as well as the decrease of cell length in response to ryanodine and IP3, were not affected. Heparin significantly blocked Ca2+ efflux and contraction in response to ACh in both conditions. ACh-stimulated accumulation of IP3 and the binding of [3H]ryanodine to its receptors were not altered by inflammation. Ruthenium red partially inhibited the response to ACh in normal and inflamed states. We conclude that the canine colonic circular muscle cells utilize Ca2+ influx through L-type channels as well as Ca2+ release from the ryanodine- and IP3-sensitive stores to contract. Inflammation impairs Ca2+ influx through L-type channels, but it may not affect intracellular Ca2+ release. The impairment of Ca2+ influx may contribute to the suppression of circular muscle contractility in the inflamed state.

Role of HERG-like K+ currents in opossum esophageal circular smooth muscle

1999 ◽  
Vol 277 (6) ◽  
pp. C1284-C1290 ◽  
Author(s):  
Hamid I. Akbarali ◽  
Hemant Thatte ◽  
Xue Dao He ◽  
Wayne R. Giles ◽  
Raj K. Goyal
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Smooth Muscle Cells ◽  
Single Cells ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Resting Membrane Potential ◽  
Channel Blockers ◽  
Circular Smooth Muscle ◽  
Inward Currents

An inwardly rectifying K+ conductance closely resembling the human ether-a-go-go-related gene (HERG) current was identified in single smooth muscle cells of opossum esophageal circular muscle. When cells were voltage clamped at 0 mV, in isotonic K+ solution (140 mM), step hyperpolarizations to −120 mV in 10-mV increments resulted in large inward currents that activated rapidly and then declined slowly (inactivated) during the test pulse in a time- and voltage- dependent fashion. The HERG K+ channel blockers E-4031 (1 μM), cisapride (1 μM), and La3+ (100 μM) strongly inhibited these currents as did millimolar concentrations of Ba2+. Immunoflourescence staining with anti-HERG antibody in single cells resulted in punctate staining at the sarcolemma. At membrane potentials near the resting membrane potential (−50 to −70 mV), this K+ conductance did not inactivate completely. In conventional microelectrode recordings, both E-4031 and cisapride depolarized tissue strips by 10 mV and also induced phasic contractions. In combination, these results provide direct experimental evidence for expression of HERG-like K+ currents in gastrointestinal smooth muscle cells and suggest that HERG plays an important role in modulating the resting membrane potential.

Electrical coupling of longitudinal and circular intestinal muscle

1984 ◽  
Vol 246 (5) ◽  
pp. G618-G626 ◽  
Author(s):  
L. Elden ◽  
A. Bortoff
Keyword(s):  
Longitudinal Muscle ◽  
Electrical Coupling ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Methyl Blue ◽  
Current Spread ◽  
Hyperpolarizing Response ◽  
Partition Method ◽  
High Degree ◽  
Longitudinal Layer

Space constants (lambda) were determined for longitudinal-circular muscle strips of cat jejunum by the partition method. Pulses of hyperpolarizing current spread along the major axes of circular muscle cells. In the absence of electrical coupling lambda measured from the longitudinal side of the strips should have been approximately 20 times shorter than lambda measured from the circular side. Median values were found to be statistically the same, 2.4 mm for the longitudinal side (n = 13) and 2.9 mm for the circular (n = 25). Methyl blue, iontophoretically injected into cells on the longitudinal side after recording large hyperpolarizing responses, was found in muscle cells located superficially in the longitudinal layer. The radial lambda for longitudinal muscle, determined from the change in magnitude of the hyperpolarizing response as the microelectrode was advanced through the layer, was 0.27 mm. This is too large to cause differences in depth of recording to significantly affect the circumferential lambda in this layer. These data provide evidence for a high degree of electrical coupling between the two muscle layers of cat jejunum.

Dual regulation of L-type Ca2+ channels by serotonin 2 receptor stimulation in vascular smooth muscle cells

1995 ◽  
Vol 268 (2) ◽  
pp. H544-H549 ◽  
Author(s):  
Y. Hirakawa ◽  
T. Kuga ◽  
S. Kobayashi ◽  
H. Kanaide ◽  
A. Takeshita
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Cell Voltage ◽  
Muscle Cells ◽  
Receptor Stimulation ◽  
Aortic Smooth Muscle ◽  
A 23187 ◽  
Inositol 1,4,5 Trisphosphate ◽  
Voltage Dependent ◽  
Ca2 Channels

The purpose of the present study was to investigate regulation of voltage-dependent Ca2+ channels by serotonin in rat aortic smooth muscle cells in primary culture. L- and T-type Ca2+ currents (ICa) were recorded using the whole cell voltage-clamp method. Without pretreatment, in 25 of 30 cells examined, 10 microM serotonin decreased L-type ICa to various extents (-14 to -72%). However, in the remaining five cells, serotonin increased L-type ICa 21 +/- 4%. Thus, in 30 cells, serotonin decreased L-type ICa an average of 22 +/- 5%. In the presence of intracellular heparin (100 micrograms/ml), a blocker of inositol 1,4,5-trisphosphate binding to its receptor, serotonin increased L-type ICa in all cells 29 +/- 3% (n = 6). When stored Ca2+ was depleted by pretreatment either with 20 microM ryanodine and 20 mM caffeine or with 100 nM A-23187, serotonin also increased L-type ICa in all cells 30 +/- 5 (n = 4) or 37 +/- 5% (n = 12), respectively. In the presence of heparin, the serotonin-induced increase of L-type ICa was prevented by 100 nM staurosporine (2 +/- 3%; n = 6, P < 0.01). The serotonin-induced decrease of L-type ICa was significantly augmented by 100 nM staurosporine (-43 +/- 10%; n = 5). Phorbol 12,13-dibutylate (PDBu; 1 microM) increased L-type ICa 29 +/- 3% (n = 6), and serotonin did not further increase L-type ICa after its potentiation by PDBu.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential inflammatory modulation of canine ileal longitudinal and circular muscle cells

1999 ◽  
Vol 277 (2) ◽  
pp. G341-G350 ◽  
Author(s):  
Xuan-Zheng Shi ◽  
Sushil K. Sarna
Keyword(s):  
Receptor Antagonist ◽  
Muscarinic Receptor ◽  
Longitudinal Muscle ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Muscle Contractions ◽  
Receptor Subtypes ◽  
Relative Role

The aim of this study was to identify the subtypes of muscarinic receptors that mediate in vivo and in vitro canine ileal longitudinal muscle contractions and whether their role is modulated by inflammation. Previous studies have reported that circular muscle contractions are suppressed in ileal inflammation induced by mucosal exposure to ethanol and acetic acid. We found that inflammation had no significant effect on in vivo and in vitro spontaneous or muscarinic receptor-mediated contractions of the longitudinal muscle. The longitudinal muscle contractions were mediated primarily by the M3 receptor subtype. However, the IC50 of the M2 receptor antagonist methoctramine was only 10 times greater than that of the M3 receptor antagonist 4-DAMP in the longitudinal muscle, whereas it was 224 times greater in the circular muscle. M2receptor-coupled decrease of intracellular cAMP occurred in the longitudinal but not in the circular muscle from the normal ileum. Inflammation did not alter this coupling in the longitudinal muscle but established it in the circular muscle. In conclusion, M2 receptors may play a greater role in the mediation of longitudinal muscle contractions than circular muscle contractions. Inflammation does not alter the contractility or the relative role of muscarinic receptor subtypes in longitudinal muscle cells. However, it modulates the M2 receptor coupling to adenylate cyclase in the circular muscle.

Regulation of Ca2+-release-activated Ca2+ current (Icrac) by ryanodine receptors in inositol 1,4,5-trisphosphate-receptor-deficient DT40 cells

2001 ◽  
Vol 360 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Kirill KISELYOV ◽  
Dong Min SHIN ◽  
Nikolay SHCHEYNIKOV ◽  
Tomohiro KUROSAKI ◽  
Shmuel MUALLEM
Keyword(s):  
Time Course ◽  
Ryanodine Receptors ◽  
Cell Types ◽  
Ruthenium Red ◽  
General Mechanism ◽  
Inward Rectification ◽  
Wild Type ◽  
Ip3 Receptor ◽  
Inositol 1,4,5 Trisphosphate ◽  
Dt40 Cells

Persistence of capacitative Ca2+ influx in inositol 1,4,5-trisphosphate (IP3) receptor (IP3R)-deficient DT40 cells (DT40IP3R-/−) raises the question of whether gating of Ca2+-release activated Ca2+ current (Icrac) by conformational coupling to Ca2+-release channels is a general mechanism of gating of these channels. In the present work we examined the properties and mechanism of activation of Icrac Ca2+ current in wild-type and DT40IP3R-/− cells. In both cell types passive depletion of internal Ca2+ stores by infusion of EGTA activated a Ca2+ current with similar characteristics and time course. The current was highly Ca2+-selective and showed strong inward rectification, all typical of Icrac. The activator of ryanodine receptor (RyR), cADP-ribose (cADPR), facilitated activation of Icrac, and the inhibitors of the RyRs, 8-N-cADPR, ryanodine and Ruthenium Red, all inhibited Icrac activation in DT40IP3R-/− cells, even after complete depletion of intracellular Ca2+ stores by ionomycin. Wild-type and DT40IP3R-/− cells express RyR isoforms 1 and 3. RyR levels were adapted in DT40IP3R-/− cells to a lower RyR3/RyR1 ratio than in wild-type cells. These results suggest that IP3Rs and RyRs can efficiently gate Icrac in DT40 cells and explain the persistence of Icrac gating by internal stores in the absence of IP3Rs.

Electrotonic current spread in colonic smooth muscle

1988 ◽  
Vol 254 (5) ◽  
pp. G702-G710 ◽  
Author(s):  
J. D. Huizinga ◽  
E. Chow
Keyword(s):  
Membrane Potential ◽  
Slow Wave ◽  
Longitudinal Muscle ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Human Colon ◽  
Membrane Properties ◽  
Electrotonic Coupling ◽  
Current Spread ◽  
Space Constant

Current-induced changes in the membrane potential (electrotonic potentials) were measured intracellularly. The electrotonic potentials were seen to decay exponentially over many cells, suggesting electrotonic current spread. The characteristics of the electrotonic current spread were used to determine passive membrane properties of both circular and longitudinal muscle cells of human and dog colon. Electrotonic current spread was first determined along the long axes of the cells. The space constant of the circular muscle of human colon was 2.14 mm and that of the longitudinal muscle was 1.63 mm. The space constants for the dog colon were similar. The value for the time constant of dog colon circular muscle was 160 ms, whereas much higher time constants, averaging between 500 and 800 ms, were recorded from dog longitudinal muscle and both human colon muscle layers. These data suggest good electrotonic coupling in all tissues studied, along the long axes of the cells. They further suggest a relatively high membrane resistance and junctional resistance in the longitudinal muscle. Electrotonic coupling along the short axes of circular muscle cells, along the long axis of the colon, was studied in the dog. The space constant was 0.43 mm, suggesting a relatively high resistance to current flow along the short axes of the cells. In addition, along the short axes of the cells from the submucosa to the myenteric plexus side (i.e., in radial direction) a gradient was observed in resting membrane potential, slow-wave amplitude, and rate of rise of the slow-wave upstroke.(ABSTRACT TRUNCATED AT 250 WORDS)

Phosphoinositide metabolism in intestinal smooth muscle: preferential production of Ins(1,4,5)P3 in circular muscle cells

1991 ◽  
Vol 261 (6) ◽  
pp. G945-G951 ◽  
Author(s):  
K. S. Murthy ◽  
G. M. Makhlouf
Keyword(s):  
Longitudinal Muscle ◽  
Inositol Phosphates ◽  
Circular Muscle ◽  
Muscle Cells ◽  
Phospholipid Metabolism ◽  
Exchange Chromatography ◽  
Phosphoinositide Metabolism ◽  
Maximal Increase ◽  
Metabolic Pattern ◽  
Inositol Phospholipid

The pattern of inositol phospholipid metabolism in response to stimulation by contractile agonists was examined in muscle cells isolated separately from circular and longitudinal muscle layers of guinea pig intestine. Addition of cholecystokinin octapeptide (CCK-8) to circular muscle cells caused a prompt decrease in phosphatidylinositol 4,5-bisphosphate (PtdInsP2) (50 +/- 6%) and PtdInsP (38 +/- 9%), which reached a nadir in 15 s. Addition of CCK-8 to longitudinal muscle cells caused a decrease in PtdInsP only (42 +/- 6%); PtdInsP2 levels, which were three times lower in this cell type, did not change throughout the period of stimulation. Hydrolysis of PtdInsP2 in circular muscle cells was accompanied by a concentration-dependent increase in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]; the maximal increase was 10 to 16 times greater in circular muscle cells as measured by radioreceptor assay and by ion-exchange chromatography. The small amounts of InsP3 produced in longitudinal muscle cells did not result from more rapid degradation, since the rates of disappearance of exogenous Ins(1,4,5)P3 in both cell types were similar. Within 5 s after addition of CCK-8, InsP3 comprised 73 +/- 5% of total inositol phosphates produced in circular muscle cells and 2.0 +/- 0.2% in longitudinal muscle cells. These divergent patterns indicate that inositol phospholipid metabolism is channeled toward generation of InsP3 in circular muscle cells only; the metabolic pattern in these cells parallels the selective presence of high-affinity InsP3 receptors and the pattern of intracellular Ca2+ mobilization.

scholarly journals Functional Ca2+ Channels between Channel Clusters are Necessary for the Propagation of IP3R-Mediated Ca2+ Waves

2019 ◽  
Vol 24 (2) ◽  
pp. 61
Author(s):  
Estefanía Piegari ◽  
Silvina Ponce Dawson
Keyword(s):  
Signal Propagation ◽  
Cell Types ◽  
Quantitative Comparison ◽  
Preliminary Conclusion ◽  
Qualitative Comparison ◽  
Excitable System ◽  
Inositol 1,4,5 Trisphosphate ◽  
Intracellular Ca ◽  
Spatio Temporal ◽  
Ca2 Channels

The specificity and universality of intracellular Ca 2 + signals rely on the variety of spatio-temporal patterns that the Ca 2 + concentration can display. Ca 2 + release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP 3 Rs) is key for this variety. The opening probability of IP 3 Rs depends on the cytosolic Ca 2 + concentration. All of the dynamics are then well described by an excitable system in which the signal propagation depends on the ability of the Ca 2 + released through one IP 3 R to induce the opening of other IP 3 Rs. In most cell types, IP 3 Rs are organized in clusters, i.e., the cytosol is a “patchy” excitable system in which the signals can remain localized (i.e., involving the release through one or more IP 3 Rs in a cluster), or become global depending on the efficiency of the Ca 2 + -mediated coupling between clusters. The spatial range over which the signals propagate determines the responses that the cell eventually produces. This points to the importance of understanding the mechanisms that make the propagation possible. Our previous qualitative comparison between experiments and numerical simulations seemed to indicate that Ca 2 + release not only occurs within the close vicinity of the clearly identifiable release sites (IP 3 R clusters) but that there are also functional IP 3 Rs in between them. In this paper, we present a quantitative comparison between experiments and models that corroborate this preliminary conclusion. This result has implications on how the Ca 2 + -mediated coupling between clusters works and how it can eventually be disrupted by the different Ca 2 + trapping mechanisms.

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