Biomedical subjects

S Wray

Publications and source records attributed to S Wray.

At least 19 recordsLinked to original sources

PubMed1999

The relationship between the action potential, intracellular calcium and force in intact phasic, guinea-pig uretic smooth muscle.

1. We investigated the relationship between the action potential, Ca2+ and phasic force in intact guinea-pig ureter, following physiological activation. 2. The action potential elicited a Ca2+ transient consisting of three components: a fast increment, associated with the first action potential spike, a slower increment, associated with subsequent spikes and the initial part of the plateau component, and a steady-state phase associated with the plateau. 3. Prolongation of the plateau, by agonists, prolonged the third component of the Ca2+ transient and increased force amplitude and duration. 4. The force-Ca2+ relationship during phasic contractions showed hysteresis; more force was produced as Ca2+ declined than when it rose. Paired pulse stimuli suggested that the delay between Ca2+ and force was not due to mechanical properties. Wortmannin, which has been shown to selectively inhibit force and myosin light chain (MLC) phosphorylation in the guinea-pig ureter, did not affect electrical activity or Ca2+ but significantly increased the delay, suggesting that myosin phosphorylation is a major contributor to it. 5. Prolongation of the duration of the [Ca2+]i transient, at unchanged amplitude, increased force. The rise of [Ca2+]i did not limit the rate of contraction. Slowing of the rate of [Ca2+]i rise abolished the hysteresis between Ca2+ and force. 6. Cooling reduced force, increased the delay and hysteresis between Ca2+ and force, but did not affect the rate of rise of Ca2+. The reduction in force could be compensated, by increasing the duration of the Ca2+ transient. 7. We suggest that in vivo, steady-state force-Ca2+ relationships are not applicable in phasic smooth muscles. Furthermore, agonists increase force mainly by prolonging the action potential, which increases the duration of the [Ca2+] signal.

Action Potentials↗

PubMed1999

The role of the sarcoplasmic reticulum as a Ca2+ sink in rat uterine smooth muscle cells.

1. The mechanisms responsible for removing calcium ions from the cytoplasm were investigated in single rat uterine myocytes using indo-1. 2. Trains of depolarizing voltage-clamp pulses increased [Ca2+]i. The rate of decay of [Ca2+]i was slowed by inhibition of the sarcoplasmic reticulum (SR) Ca2+-ATPase with cyclopiazonic acid (CPA). However, if the sarcolemmal Na+-Ca2+ exchanger and Ca2+-ATPase were inhibited then recovery of [Ca2+]i was abolished showing that the SR Ca2+-ATPase alone cannot produce decay of [Ca2+]i. 3. In another series of experiments, Ca2+ release from the SR was induced with carbachol in a Ca2+-free solution. Under these conditions responses to repeated applications of carbachol could be obtained. In the presence of CPA, however, only the first application was effective. This suggests that the SR Ca2+-ATPase sequesters a significant amount of Ca2+ into the SR. 4. CPA slowed the rate of decay of [Ca2+]i following carbachol addition by > 50 %. Again, however, after a brief transient fall, decay was abolished when the Na+-Ca2+ exchanger and sarcolemmal Ca2+-ATPase were inhibited. 5. These data show that, although the SR Ca2+-ATPase contributes to the decay of [Ca2+]i, it cannot function effectively in the absence of Ca2+ removal from the cell. These data are discussed in the context of the superficial buffer barrier model in which Ca2+ is taken up into the SR and then released very close to sarcolemmal Ca2+ extrusion sites, i.e. the SR acting in series with the surface membrane extrusion mechanisms. We also suggest that the amount of filling of the SR influences the rate of Ca2+ removal.

Animals↗

PubMed1999

In vivo pH and metabolite changes during a single contraction in rat uterine smooth muscle.

1. We have used 31P NMR spectroscopy to measure metabolites and pHi at three periods during a phasic contraction of the uterus, in vivo, to determine whether they change as a consequence of contraction. The regular uterine contractions were recorded via a balloon catheter in the uterine lumen. Each phasic contraction was divided into three parts: the period between contractions (rest), the development of force (up) and the relaxation of force (down). The NMR data were summed separately from each of these three periods over 20-40 successive contractions. 2. Significant changes in ATP, phosphocreatine (PCr) and inorganic phosphate (Pi) occurred during the contraction. [ATP] fell from 2.0 to 1.6 mM and [PCr] from 2.6 to 2.0 mM during the up period, while [Pi] increased from 2.2 to 2.8 mM. Recovery of ATP and PCr occurred during the relaxation part of the contraction, whereas Pi did not fully recover until the contraction was complete. 3. Significant acidification from pH 7.28 +/- 0.02 at rest to 7.16 +/- 0.02, occurred with contraction. This acidification is greater than that previously reported for in vitro uterine preparations. Measurements of uterine blood flow show that it decreased with contraction. Therefore, ischaemia, in addition to the metabolic consequences of contraction, may account for the larger acidification observed in vivo. 4. Lowering pHi in an in vitro uterine preparation by a similar level to that found in vivo produced a significant reduction of the phasic contractions. Thus we propose that these changes, especially the fall in pHi during force development, feed back negatively on the contraction to limit its strength, and may help prevent uterine ischaemia and fetal hypoxia during labour.

Adenosine Triphosphate↗

PubMed1999

The effect of cyclopiazonic acid on excitation-contraction coupling in guinea-pig ureteric smooth muscle: role of the sarcoplasmic reticulum.

1. We have investigated the effect of cyclopiazonic acid (CPA), an inhibitor of the sarcoplasmic reticulum (SR) Ca2+-ATPase on excitation-contraction (EC) coupling in guinea-pig ureter, by measuring membrane currents, action potentials, intracellular [Ca2+] and force. 2. CPA (20 micrometers) significantly enhanced the amplitude and duration of phasic contractions of ureteric smooth muscle associated with action potentials. This was accompanied by an increase in the duration of the intracellular Ca2+ transient in intact tissue and single cells but not their amplitude. However, CPA also slowed the rate of rise, and fall, of the force 1|1|Phiand1Phi Ca2+ transients. 3. Membrane potential recordings showed that CPA produced a small depolarization and a large increase in the duration of the plateau phase of the action potential. 4. Patch-clamp studies showed marked inhibition of outward potassium current in the presence of CPA and an inhibition of spontaneous transient outward currents (STOCs). CPA had no effect on inward Ca2+ current. 5. These data suggest that the SR plays a major role in modulating the excitability of the ureter, particularly via curtailing the action potential duration. This in turn will shorten the Ca2+ transient and decrease force. This negative action on developed force predominates over any small role it may play in initiating force in the guinea-pig ureter.

Animals↗

PubMed1999

Intracellular calcium stores and agonist-induced contractions in isolated human myometrium.

OBJECTIVE: We hypothesized that the release of calcium from intracellular stores contributes to the contractions produced by the agonists oxytocin, carbachol, and prostaglandin F(2 )(alpha ) in human myometrium. STUDY DESIGN: Strips of myometrium were obtained at cesarean section and hysterectomy. The strips were loaded with the calcium-sensitive dye Indo-1 to enable simultaneous measurement of tension and intracellular calcium levels. Agonist-induced responses in the presence and absence of extracellular calcium were studied. RESULTS: Strips of myometrium were obtained from 48 women not in labor undergoing cesarean section and 6 women not pregnant undergoing hysterectomy. An increase in intracellular calcium level after agonist stimulation invariably preceded an increase in tension. Intracellular calcium level returned to baseline before myometrial relaxation. Oxytocin, carbachol, and prostaglandin F(2)(alpha) all gave both force and intracellular calcium responses in the absence of extracellular calcium, although both these responses were only 26% to 40% of the maximal response when extracellular calcium was present. CONCLUSIONS: Release of calcium from internal stores induced by oxytocin, carbachol, and prostaglandin F(2)(alpha) may contribute to agonist-induced myometrial force production.

Adult↗

PubMed1999

Luteinizing hormone-releasing hormone quantified in tissues and slice explant cultures of postnatal rat hypothalami.

LH-releasing hormone (LHRH) peptide from postnatal rat preoptic area (POA)/hypothalamic tissues in vivo and slice explant cultures maintained in vitro was quantitated using an enzyme-linked immunosorbant assay. Moreover, messenger RNA (mRNA) copy number was calculated in LHRH neurons maintained in culture using in situ hybridization histochemistry with autoradiographic film analysis. POA/hypothalami from postnatal day 5-6 pups averaged 1250 pg of LHRH, with approximately 28% of peptide residing within rostral tissues where most LHRH perikarya reside. Explant cultures maintained 18 days in vitro contained 30.4-92.0 pg/slice with a whole animal total of 244.8 pg. Considering cell numbers in vivo and in vitro, LHRH neurons in whole animal produce 1.0 pg of LHRH/cell, whereas those in culture average 2.0 pg/cell. Furthermore, LHRH mRNA copies/cell in organotypic culture was estimated conservatively at 1410 copies/cell, a relatively high number. This work shows that, compared with whole animal, cultures have substantial LHRH stores, indicating maturation of synthetic activity and/or formation of new terminals in vitro. High LHRH mRNA copy number also suggests a high rate of peptide biosynthesis. Our analysis, demonstrating the dynamic potential of LHRH neurons, suggests that subtle changes in LHRH mRNA expression in all cells or a subpopulation can dramatically alter the LHRH system biosynthetic capacity.

Animals↗

PubMed1998

Properties of voltage-activated [Ca2+]i transients in single smooth muscle cells isolated from pregnant rat uterus.

1. The intracellular calcium concentration ([Ca2+]i) was measured at 35 degrees C using the fluorescent indicator indo-1 in patch-clamped, single uterine myocytes from pregnant rats to investigate the relationship between depolarization, Ca2+ current (ICa) and [Ca2+]i. 2. Membrane depolarization activated ICa and produced a [Ca2+]i transient. The rapid increase in [Ca2+]i occurred at the same time as the inward ICa. Both ICa and the increase in [Ca2+]i were abolished by nifedipine (10 microM). 3. When the membrane potential was held at -80 mV the threshold depolarization for an increase in [Ca2+]i was about -55 to -50 mV. As the magnitude of the depolarization was increased to about 0 mV there was an increase in the size of both ICa and the increase in [Ca2+]i. As the magnitude of the depolarization was further increased both ICa and the [Ca2+]i increase declined. 4. When the depolarizing pulses were applied at 3 Hz to mimic normal action potentials then the individual [Ca2+]i transients did not fully relax and a tetanic rise of [Ca2+]i was observed. Under these conditions, there was not a simple relationship between the magnitude of the Ca2+ response and Ca2+ entry. When pairs of depolarizing pulses were applied, the increase in [Ca2+]i produced by the second pulse was larger (in relation to the magnitude of the L-type Ca2+ current) than that produced by the first pulse. This facilitation was abolished by both ryanodine and cyclopiazonic acid suggesting a role for release from intracellular stores. 5. We conclude that the L-type Ca2+ current is the major source of Ca2+ ions entering the cell to produce the [Ca2+]i transient on depolarization. The magnitude of the increase in [Ca2+]i may, however, be amplified by Ca2+-induced Ca2+ release.

Animals↗

PubMed1998

Contribution of sarcoplasmic reticular calcium to smooth muscle contractile activation: gestational dependence in isolated rat uterus.

1. The contribution of Ca2+ released from the sarcoplasmic reticulum (SR) to smooth muscle contractile activation remains poorly understood. By simultaneously monitoring cytosolic [Ca2+] ([Ca2+]i) and force in isolated rat uterine smooth muscle, we report the influence of SR Ca2+ release on contractility during conditions (a) of altered SR Ca2+ homeostasis and (b) where the only source of activating Ca2+ was derived from the SR. 2. In myometria of non-pregnant rats, ryanodine (1-50 microM), a modulator of calcium-induced calcium release (CICR), had no effect on the spontaneous [Ca2+]i or force transients. However, depletion of SR Ca2+ by inhibiting the SR Ca2+-ATPase (with cyclopiazonic acid (CPA), 20 microM) resulted in an enhancement of spontaneous [Ca2+]i and force transients. 3. In myometria of pregnant rats, although ryanodine had no effect in 40% of tissues studied it produced a small but significant enhancement of the integrated spontaneous [Ca2+]i and force transient in 60% of cases. The potentiating effects of CPA were enhanced in myometria of pregnant rats compared with non-pregnant rats, often resulting in maintained [Ca2+]i increases and contraction. 4. In zero external Ca2+, agonist-induced SR Ca2+ release resulted in transient increases in [Ca2+]i and force. The magnitude of these agonist-induced [Ca2+]i and force changes were significantly enhanced in myometria of pregnant rats. No evidence for agonist-induced Ca2+-independent force production was observed. 5. These results indicate that CICR plays little role in SR Ca2+ release from the myometrium, and that there are gestational-dependent alterations in the ability of SR Ca2+ mobilization to contribute to contractile activation. The implications of these findings for the co-ordination of myometrial [Ca2+]i signalling and contractility are discussed.

Animals↗

PubMed1998

Hypoxia and smooth muscle function: key regulatory events during metabolic stress.

Hypoxia rapidly reduces force in many smooth muscles and we review recent data that shed light on the mechanisms involved. As many regulated cellular processes are integrated to co-ordinate smooth muscle contractility, the processes responsible for decreased force output with altered metabolism are also likely to be many, acting in concert, rather than the actions of one altered parameter. Nevertheless the aim of this study is to elucidate the hierarchical series of events that contribute to reduced smooth muscle force production during altered metabolism. We conclude that in many phasic smooth muscles the decrease in force can be attributed to impaired electro-mechanical coupling whereby the Ca2+ transient is reduced. A direct effect of hypoxia on the Ca2+ channel may be of key importance. In tonic vascular smooth muscles KATP channels may also play a role in the integrated functional responses to hypoxia. There are also many examples of force being reduced, in tonically activated preparations, without a fall in steady-state Ca2+; indeed it usually increases. We examine the roles of altered [ATP], pH, myosin phosphorylation, inorganic phosphate and proteolytic activity on the [Ca2+]-force relationship during hypoxia. We find no defining force-inhibitory role for any one factor acting alone, and suggest that force most probably falls as a result of the combination of myriad factors.

Animals↗

PubMed1998

GABA inhibits migration of luteinizing hormone-releasing hormone neurons in embryonic olfactory explants.

During development, a subpopulation of olfactory neurons transiently expresses GABA. The spatiotemporal pattern of GABAergic expression coincides with migration of luteinizing hormone-releasing hormone (LHRH) neurons from the olfactory pit to the CNS. In this investigation, we evaluated the role of GABAergic input on LHRH neuronal migration using olfactory explants, previously shown to exhibit outgrowth of olfactory axons, migration of LHRH neurons in association with a subset of these axons, and the presence of the olfactory-derived GABAergic neuronal population. GABAA receptor antagonists bicuculline (10(-5) M) or picrotoxin (10(-4) M) had no effect on the length of peripherin-immunoreactive olfactory fibers or LHRH cell number. However, LHRH cell migration, as determined by the distance immunopositive cells migrated from olfactory pits, was significantly increased by these perturbations. Addition of tetrodotoxin (10(-6) M), to inhibit Na+-transduced electrical activity, also significantly enhanced LHRH migration. The most robust effect observed was dramatic inhibition of LHRH cell migration in explants cultured in the presence of the GABAA receptor agonist muscimol (10(-4) M). This study demonstrates that GABAergic activity in nasal regions can have profound effects on migration of LHRH neurons and suggests that GABA participates in appropriate timing of LHRH neuronal migration into the developing brain.

Animals↗

PubMed1998

The mechanism of Ca2+ release from the SR of permeabilised guinea-pig and rat ureteric smooth muscle.

Recent work has indicated that there is a major difference in the Ca2+ store of smooth muscle from rat and guinea-pig ureter; with the rat store being agonist-sensitive but ryanodine insensitive and the guinea-pig store being ryanodine sensitive but agonist insensitive [Th. V. Burdyga, M.J. Taggart, S. Wray, J. Physiol. 489 (1995) 327-335]. We have therefore examined directly the mechanism of Ca2+ release from the internal Ca2+ store (SR). Following permeabilisation with alpha-toxin or beta-escin the SR was Ca(2+)-loaded before application of carbachol or caffeine. Only carbachol evoked a transient contraction in rat ureter. The carbachol-induced contraction was blocked by heparin and cyclopiazonic acid (CPA) but not ryanodine. Only caffeine produced contraction in guinea-pig ureter, and this was blocked by ryanodine. Direct application of IP3 caused a small transient contraction in rat but not guinea-pig ureter. We conclude that rat ureter possesses only an IP3 sensitive store while guinea-pig ureter only has a ryanodine sensitive store.

Animals↗

PubMed1998

The effects of metabolic inhibition on force, Ca2+ and pHi in guinea-pig ureteric smooth muscle.

The effect of metabolic inhibition on the contractile function of adult guinea-pig ureter has been investigated. Strips of ureteric smooth muscle were loaded with Indo-1 or SNARF to measure intracellular [Ca2+] ([Ca2+]i) or pH (pHi) simultaneously with force. Inhibiting oxidative phosphorylation with cyanide rapidly reduced phasic contractions and the associated Ca2+ transients, after initial transient increases. The effects of cyanide were reversible and related to the amount of contractile activity undertaken. Inhibition of glycolysis with iodoacetate abolished all force. In high-K+-depolarised preparations, cyanide reduced the tonic contraction, but this was not accompanied by a reduction in [Ca2+]i, suggesting a desensitisation of the myofilaments. Cyanide produced a fall in pHi, which may underlie the initial transient increase in force. These data suggest that metabolic inhibition reduces force in the ureter by affecting both excitation and hence the Ca2+ transient, and at the myofilaments to reduce their sensitivity to Ca2+. Thus when oxidative metabolism is impaired contractile dysfunction may arise in the ureter.

Animals↗

PubMed1998

The effects of changing intracellular pH on calcium and potassium currents in smooth muscle cells from the guinea-pig ureter.

Guinea-pig ureteric smooth muscle is unusual in that intracellular acidification increases and alkalinization decreases force production. To help elucidate the mechanism underlying these effects on force we have investigated the effects of changing intracellular pH on both calcium and potassium currents in single cells isolated from the guinea-pig ureter to determine their possible role in force development. Depolarization to +40 mV resulted in a fast transient outward current which was inhibited by 4-aminopyridine but not tetraethylammonium. Intracellular alkalinization (20 mM trimethylamine) increased this current to 179 +/- 24% of the control and resulted in the development of a slowly activating large outward current which was inhibited by tetraethylammonium and washout. Acidification (40 mM sodium butyrate) decreased the fast transient outward current to 58 +/- 3% of the control and did not produce a slowly activating current. When potassium was replaced by caesium in the pipette solution, depolarization to 0 mV resulted in an inward calcium current which was abolished by nifedipine. Intracellular alkalinization increased this current to 126 +/- 11% of the control whereas acidification had the opposite effect, decreasing it to 55 +/- 10%. Furthermore, current-clamp experiments showed that intracellular alkalinization inhibited the amplitude of the action potential, therefore decreasing excitability of the cell. From our results, we suggest that the predominant effects of intracellular pH on force production in the guinea-pig ureter are mediated via the modulation of outward potassium currents (thereby reducing excitability of the tissue) rather than the effects on the inward calcium current.

Animals↗

PubMed1998

Intracellular Na+ measurements in smooth muscle using SBFI--changes in [Na+], Ca2+ and force in normal and Na(+)-loaded ureter.

Our understanding of the control and effects of intracellular [Na+] ([Na+]i) in intact smooth muscle is limited by the lack of data concerning [Na+]i. The initial aim of this work was therefore to investigate the suitability of using the Na+-sensitive fluorophore SBFI in intact smooth muscle. We find this to be a good method for measuring [Na+]i in ureteric smooth muscle. Resting [Na+]i was found to be around 10 mM and rose to 25 mM when the Na+-K+-ATPase was inhibited by ouabain. This relatively low [Na+]i in the absence of Na+-K+-ATPase suggests that other cellular processes, such as Na+-Ca2+ exchange, play a role in maintaining [Na+]i under these conditions. Simultaneous measurements of [Na+]i or [Ca2+] i and force showed that Na+-Ca2+ exchange can play a functional role in ureteric smooth muscle. We found that the greater the driving force for Na+ exit and hence Ca2+ entry, the larger the contraction. In addition the Na+-Ca2+ exchanger activity under these conditions was found to be pH sensitive: acidification reduced the contraction and concomitant changes in [Ca2+] and [Na+]i. We conclude that SBFI is a useful method for monitoring [Na] in smooth muscle and that Na+-Ca2+ exchange may play a functional role in the ureter.

Animals↗

PubMed1998

Developmental changes in intracellular pH buffering power in smooth muscle.

Intracellular pH (pHi) is known to modulate contraction. Neonatal tissues can differ from adult tissue in contractile response to stimuli known to alter pHi e.g. hypoxia. Changes of pH are attenuated by buffering, thus any difference in buffering power (beta) between tissues could affect their functional response to pHi perturbation. Similarly the extent to which any extracellular pH (pHo) alteration is transmitted into a pHi change will also influence function. We have therefore determined the intrinsic beta and effect of pHo change on pHi in neonatal and adult ureteric, uterine and gastric smooth muscles using the pH-sensitive fluorophore carboxy-SNARF. beta was found to be similar in the three adult tissues, but there were significant differences between neonatal tissues. In contrast, we found little difference in the amount of pHi change produced by pHo change between neonatal and adult tissues from the same smooth muscle, but a difference between smooth muscles. These data highlight significant differences between smooth muscles and their developmental state, which may contribute to different degrees of protection when pH is perturbed.

Animals↗

PubMed1998

The in vivo relationship between blood flow, contractions, pH and metabolites in the rat uterus.

Little is known about the relationship between smooth muscle contractile activity and its blood supply. We have therefore investigated this in the rat uterus, using laser-Doppler flow measurement and intra-uterine pressure recordings. We found an inverse linear relationship between flow and contractile activity. There was no evidence for a critical level of flow, above which function is maintained and below which it declines; even small reductions in blood flow decreased uterine force. Force was rapidly restored upon reperfusion. Reactive hyperaemia was absent from all but 6 of the 41 preparations studied. We used 31P nuclear magnetic resonance (NMR) spectroscopy to measure concentrations of adenosine triphosphate (ATP), phosphocreatine (PCr), inorganic phosphate (Pi) and intracellular pH (pHi) simultaneously with force and flow. Reductions in flow were associated with significant reductions in [ATP], [PCr] and pHi, and an increase in [Pi]. These changes were related to flow significantly and linearly and their effects on force may be additive. These data show that uterine smooth muscle is closely dependent upon its blood supply for maintaining both normal force production and metabolite levels. Consequently, even small decrements in flow may have deleterious functional effects.

Adenosine Triphosphate↗

PubMed1998

Developmental and species differences in the response of the ureter to metabolic inhibition.

The effect of inhibiting oxidative phosphorylation on electrically stimulated phasic and high-K+ depolarization-induced tonic contractions in ureteric smooth muscle has been investigated. Intracellular [Ca2+] and pH were monitored fluorimetrically with simultaneous tension measurement, in adult and neonatal rat and guinea-pig ureter. Little difference was found in the response of adult or neonatal rat ureters; cyanide abolished phasic contractions and intracellular Ca2+ transients. The contractions of the adult guinea-pig ureter were also reduced by cyanide, but not as much as those of the adult rat. Neonatal guinea-pig was, however, remarkably resistant to the effects of cyanide, with force and Ca2+ transients remaining at control levels after an initial transient dip. These differences between tissues were not apparent when a high K+ concentration was used to depolarize tissues and produce maintained [Ca2+]i and force changes; cyanide reduced force but not [Ca2+]i in all preparations. Intracellular pH decreased in all preparations with inhibition of oxidative phosphorylation, but this did not correlate with changes in contraction. It is concluded that there are both species and developmental differences in the response to metabolic inhibition of the ureter which lead to differing changes in contractile activity.

Animals↗

PubMed1998

The effect of inhibition of myosin light chain kinase by Wortmannin on intracellular [Ca2+], electrical activity and force in phasic smooth muscle.

To investigate the role of myosin light chain kinase (MLCK) in phasic contractions of intact smooth muscle, we have applied Wortmannin, an MLCK inhibitor, to strips of guinea-pig ureter. Simultaneous measurements of electrical activity, intracellular [Ca2+] ([Ca2+]i) and phasic force showed that Wortmannin (1-4 microM) abolishes force with little or no change in [Ca2+]i and electrical activity. High-K+-induced force production was also abolished by Wortmannin. The effects of Wortmannin were dose dependent - at lower concentrations (100 nM) Wortmannin reduced phasic contractility by 40-50%. It also significantly increased the delay between the Ca2+ peak and force production. These data show that, in phasic smooth muscle, inhibition of MLCK causes contraction to fail, despite normal electrical activity and Ca2+ transients. Our results also indicate that Wortmannin has no secondary effects and that other means of producing force, independent of myosin phosphorylation, are negligible in this tissue. The increased lag between the rise of Ca2+ and force production when MLCK is inhibited was surprising and suggests that post-phosphorylation steps may play a larger role in the delay than was previously considered.

Action Potentials↗