scholarly journals Subcellular-membrane characterization of [3H]ryanodine-binding sites in smooth muscle

1993 ◽  
Vol 290 (1) ◽  
pp. 259-266 ◽  
Author(s):  
Z D Zhang ◽  
C Y Kwan ◽  
E E Daniel
Keyword(s):  
Smooth Muscle ◽  
Ryanodine Receptor ◽  
Binding Sites ◽  
Vas Deferens ◽  
Smooth Muscles ◽  
Rat Vas Deferens ◽  
Detailed Characterization ◽  
Release Channel ◽  
Subcellular Membrane

The plant alkaloid ryanodine, known to interact selectively with the intracellular Ca(2+)-release channel in skeletal and cardiac muscles, has been repeatedly reported to affect smooth-muscle contractile functions that are consistent with its intracellular action at the Ca(2+)-release channel sites. Direct evidence for the binding of [3H]ryanodine to smooth-muscle membranes is sparse. Following our recent detailed characterization of functional effects of ryanodine and a preliminary report on the presence of [3H]ryanodine binding sites in rat vas deferens smooth muscle, we now report in this study a detailed characterization of binding of [3H]ryanodine to smooth muscle at the subcellular-membrane level. The ryanodine receptor in rat vas deferens muscle layer is primarily of smooth-muscle origin and is localized at the subcellular membrane site that is consistent with its role as a Ca(2+)-release channel in the sarcoplasmic reticulum (SR). Ryanodine binding to its receptor is Ca(2+)-dependent, with half-maximal binding occurring within the physiologically relevant cytosolic Ca2+ concentration. It is also sensitive to many factors, including change in Mg2+ concentration, ionic strength and osmolarity across the membrane vesicles. Agents known to inhibit (Ruthenium Red, Mg2+) or enhance (caffeine, Na+, K+) the Ca(2+)-induced Ca2+ release also inhibit or enhance the binding of ryanodine. Quantitative differences in ryanodine receptors exist among smooth muscles and do not seem to parallel their SR contents. Results from the present study indicate both the need and the basis for future investigations of the functional role of the ryanodine receptor in different smooth muscles.

Effect of NO donors on protein phosphorylation in intact vascular and nonvascular smooth muscles

2001 ◽  
Vol 280 (4) ◽  
pp. H1565-H1580 ◽  
Author(s):  
James K. Hennan ◽  
Jack Diamond
Keyword(s):  
Smooth Muscle ◽  
Protein Kinase ◽  
Protein Phosphorylation ◽  
Vas Deferens ◽  
Smooth Muscles ◽  
Rat Aorta ◽  
Rat Vas Deferens ◽  
Two Dimensional Gel Electrophoresis ◽  
Relaxant Response ◽  
Underlying Mechanisms

It is generally well accepted that nitrovasodilator-induced relaxation of vascular smooth muscle involves elevation of cGMP and activation of a specific cGMP-dependent protein kinase [protein kinase G (PKG)]. However, the protein targets of PKG and the underlying mechanisms by which this kinase leads to a relaxant response have not been elucidated. Several types of smooth muscle, including rat myometrium and vas deferens, are not relaxed by sodium nitroprusside, even at concentrations that produce marked elevation of cGMP and activation of PKG. The main objective of our studies was to compare PKG-mediated protein phosphorylation in intact rat aorta, rat myometrium, and rat vas deferens using two-dimensional gel electrophoresis. In intact rat aorta, seven PKG substrates were detected during relaxation of the tissue. None of the PKG substrates identified in the rat aorta appeared to be phosphorylated in the myometrium or vas deferens after administration of various cGMP-elevating agents. Thus the failure of the rat myometrium and rat vas deferens to relax in the face of cGMP elevation and PKG activation may be due to a lack of PKG substrate phosphorylation.

Characterization of receptors for angiotensin in the rat vas deferens

1979 ◽  
Vol 57 (4) ◽  
pp. 417-423 ◽  
Author(s):  
J. Magnan ◽  
D. Regoli
Keyword(s):  
Smooth Muscle ◽  
Vas Deferens ◽  
Proteolytic Enzymes ◽  
Rabbit Aorta ◽  
Rat Vas Deferens ◽  
Converting Enzyme ◽  
Response Curves ◽  
Sympathetic Nerve Stimulation ◽  
The Right

Experiments were performed in the rat vas deferens to characterize the receptor for angiotensin mediating the potentiation of the sympathetic nerve stimulation by this peptide. For this purpose we measured the order of potency of various angiotensins, the affinity of two specific and competitive antagonists, and we compared the effects of several angiotensins in tissues desensitized by angiotensin II.The potency of natural angiotensins follows the order: ATII > ATI > ATIII the relative potency of a few analogues which resist degradation by proteolytic enzymes as well as the potency of three L-Ala analogues of ATII show similar changes as those observed in other smooth muscle preparations (e.g. the rabbit aorta).Affinity of antagonists was evaluated by measuring pA2 and is higher for [Leu8]-ATII than for [des-Asp1, Leu8]-ATII. Both antagonists appear to be competitive since they displace the dose-response curves of ATII and ATIII to the right without changing the slope of the curves. Desensitization with ATII renders the tissues insensitive to ATIII and to other angiotensins without changing the response of the tissues to substance P.ATII does not modify the action of exogenous NA on nonstimulated tissues. ATI has no direct effect since its action is completely blocked in the presence of an inhibitor of the converting enzyme (SQ. 14225), while the responses of the vas deferens to ATII and substance P are unaltered.It is concluded that the receptor for ATII in the rat vas deferens is of the same type as the receptor mediating contraction of the rabbit aorta.

Characterization of [3H]Ro 5-4864 Binding Sites in Rat Vas Deferens

1992 ◽  
Vol 58 (1) ◽  
pp. 39-45 ◽  
Author(s):  
E. Escubedo ◽  
A. Camins ◽  
C. Talaveron ◽  
J. Camarasa
Keyword(s):  
Binding Sites ◽  
Vas Deferens ◽  
Rat Vas Deferens

scholarly journals Electrophysiology of Syncytial Smooth Muscle

2019 ◽  
Vol 13 ◽  
pp. 117906951882191 ◽  
Author(s):  
Rohit Manchanda ◽  
Shailesh Appukuttan ◽  
Mithun Padmakumar
Keyword(s):  
Smooth Muscle ◽  
Action Potentials ◽  
Signal Analysis ◽  
Computational Models ◽  
Vas Deferens ◽  
Contractile Activity ◽  
Smooth Muscles ◽  
Biophysical Analysis ◽  
Recent Developments ◽  
The Many

As in other excitable tissues, two classes of electrical signals are of fundamental importance to the functioning of smooth muscles: junction potentials, which arise from neurotransmission and represent the initiation of excitation (or in some instances inhibition) of the tissue, and spikes or action potentials, which represent the accomplishment of excitation and lead on to contractile activity. Unlike the case in skeletal muscle and in neurons, junction potentials and spikes in smooth muscle have been poorly understood in relation to the electrical properties of the tissue and in terms of their spatiotemporal spread within it. This owes principally to the experimental difficulties involved in making precise electrical recordings from smooth muscles and also to two inherent features of this class of muscle, ie, the syncytial organization of its cells and the distributed innervation they receive, which renders their biophysical analysis problematic. In this review, we outline the development of hypotheses and knowledge on junction potentials and spikes in syncytial smooth muscle, showing how our concepts have frequently undergone radical changes and how recent developments hold promise in unraveling some of the many puzzles that remain. We focus especially on computational models and signal analysis approaches. We take as illustrative examples the smooth muscles of two organs with distinct functional characteristics, the vas deferens and urinary bladder, while also touching on features of electrical functioning in the smooth muscles of other organs.

Characterization of the ryanodine receptor/channel of invertebrate muscle

1998 ◽  
Vol 274 (2) ◽  
pp. R494-R502 ◽  
Author(s):  
Kerry E. Quinn ◽  
Loriana Castellani ◽  
Karol Ondrias ◽  
Barbara E. Ehrlich
Keyword(s):  
Ryanodine Receptor ◽  
Single Channel ◽  
Microscopic Analysis ◽  
Ruthenium Red ◽  
Electron Microscopic ◽  
Release Channel ◽  
Single Channel Currents ◽  
Channel Currents ◽  
Bell Shaped Curve

Electron-microscopic analysis was used to show that invertebrate muscle has feetlike structures on the sarcoplasmic reticulum (SR) displaying the typical four-subunit appearance of the calcium (Ca2+) release channel/ryanodine receptor (RyR) observed in vertebrate skeletal muscle (K. E. Loesser, L. Castellani, and C. Franzini-Armstrong. J. Muscle Res. Cell Motil. 13: 161–173, 1992). SR vesicles from invertebrate muscle exhibited specific ryanodine binding and single channel currents that were activated by Ca2+, caffeine, and ATP and inhibited by ruthenium red. The single channel conductance of this invertebrate RyR was lower than that of the vertebrate RyR (49 and 102 pS, respectively). Activation of lobster and scallop SR Ca2+ release channel, in response to cytoplasmic Ca2+ (1 nM–10 mM), reflected a bell-shaped curve, as is found with the mammalian RyR. In contrast to a previous report (J.-H. Seok, L. Xu, N. R. Kramarcy, R. Sealock, and G. Meissner. J. Biol. Chem. 267: 15893–15901, 1992), our results show that regulation of the invertebrate and vertebrate RyRs is quite similar and suggest remarkably similar paths in these diverse organisms.

The Sarcoplasmic Reticulum of Smooth Muscle Fibers

1982 ◽  
Vol 37 (5-6) ◽  
pp. 481-488 ◽  
Author(s):  
L. Raeymaekers
Keyword(s):  
Smooth Muscle ◽  
Calcium Oxalate ◽  
Striated Muscle ◽  
Smooth Muscles ◽  
Morphological Characterization ◽  
Calcium Oxalate Crystals ◽  
Additional Amount ◽  
Skinned Smooth Muscle ◽  
Ca Uptake

Abstract The ability of the sarcoplasmic (endoplasmic) reticulum (SR, ER) of smooth muscle cells to accumulate Ca was demonstrated by measuring the uptake of 45Ca in fibers which were chemically skinned with saponin, and by electron cytochemistry of the accumulated Ca. The Ca uptake was dependent on ATP and it was stimulated by oxalate, as it is the case in SR of striated muscle. Electron microscopy of the skinned smooth muscle preparations revealed the presence of calcium oxalate deposits in the reticulum. The SR vesicles were isolated from several smooth muscles. The purification was carried out by taking advantage of the density increase of the SR vesicles after loading with calcium in the presence of oxalate. Among the muscles investigated the smooth muscle of the pig stomach was found to be the most suitable and it was selected for further biochemical and morphological characterization of the SR vesicles. These vesicles, which contain calcium oxalate crystals, were able to accumulate an additional amount of Ca. The Ca uptake was supported by several energy yielding substrates. Their order of potency was ATP > dATP ≃ UTP > ITP > GTP ≃ CTP. The rate of Ca uptake was two orders of magnitude slower than that in SR of skeletal muscle. The measurement of the level of phosphorylated Ca transport intermediate showed that this difference is due to smaller number of calcium transport sites per vesicle. The distribution of intramembrane particles in freeze-fractured specimens is in agreement with this conclusion.

Ca2+ handling properties of microsomal subfractions of rat vas deferens smooth muscle

1984 ◽  
Vol 62 (1) ◽  
pp. 76-79 ◽  
Author(s):  
A. K. Grover ◽  
C. Y. Kwan
Keyword(s):  
Smooth Muscle ◽  
Concentration Dependence ◽  
Vas Deferens ◽  
Membrane Vesicles ◽  
Rat Vas Deferens ◽  
Enzyme Marker ◽  
Isopycnic Centrifugation ◽  
Stimulation Of ◽  
Ph Profiles

The rat vas deferens smooth muscle microsomes on isopycnic centrifugation gave two fractions, namely F2 (15–30% sucrose) and F3 (30–40% sucrose), with comparable ATP-dependent azide-insensitive Ca2+-uptake capacities, although these fractions differed from each other in various enzyme marker activities. The fractions F2 and F3 also show similar pH profiles for the ATP-independent and ATP-dependent Ca2+ uptake, and similar ionized Ca2+-concentration dependence for the ATP-dependent Ca2+ uptake. However, the fractions F2 and F3 differ from each other in that: (a) F3 shows higher permeability to Ca2+, and (b) F3 shows higher stimulation of the ATP-dependent Ca2+ uptake by oxalate. The F3 fraction can also be used to obtain membrane vesicles loaded with Ca2+ oxalate in the presence of ATP. However, the yield of the Ca2+ oxalate enriched fraction is too low to permit their further characterization.

Decreased density of binding sites for the Ca2+ channel antagonist [3H]isradipine after denervation of rat vas deferens

1994 ◽  
Vol 256 (3) ◽  
pp. 329-333 ◽  
Author(s):  
Aron Jurkiewicz ◽  
Simone S. Lafayette ◽  
Sandra H. Nunes ◽  
Lucienne C. Martini ◽  
Lucia G. Do Carmo ◽  
...  
Keyword(s):  
Binding Sites ◽  
Vas Deferens ◽  
Rat Vas Deferens
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