Rhythmically Contracting Research Articles

Spike of interstitial PO2 produced by a twitch in rhythmically contracted muscle.

Oxygen (O2) exchange between capillaries and muscle cells in exercising muscles is of great interest for physiology and kinesiology. However, methodical limitations prevent O2 measurements on the millisecond scale. To bypass the constraints of quasi‐continuous recording, progressive measurements of O2 partial pressure (PO2) in rhythmically contracting skeletal muscle were compiled to describe the O2 kinetics surrounding and including a single muscle contraction. Phosphorescence quenching microscopy measured PO2 in the interstitium of the rat spinotrapezius muscle. Measurements were triggered by contraction‐inducing electrical pulses. For the first 60 seconds, measurement preceeded stimulation. After 60, measurement followed with a progressive 20 ms increment. Thus, the first 60 measurements describe the overall PO2 response to electrical stimulation initiated after a 10 second rest period, while 61–100 (stroboscopic mode) were compiled into a single 800 ms profile of the PO2 transient surrounding muscle contraction. Thirty seconds of stimulated contractions decreased interstitial PO2 from a baseline of 71 ± 1.4 mmHg to an “active” steady‐state of 43 ± 1.5 mmHg. The stroboscopic mode compilation revealed an unexpected post‐contractile rise in PO2 as a 205 ms spike with a maximum amplitude of 58 ± 3.8 mmHg at 68 ms, which restored 58% of the PO2 drop from baseline. Interpretation of this phenomenon is based on classical experiments by G.V. Anrep (1935), who discovered the rapid thrust of blood flow associated with muscle contraction. In addition to the metabolic implications during exercise, the physiological impact of these PO2 spikes may grow with an increased rate of rhythmical contractions in muscle or heart.New&NoteworthyThe principal finding is a spike of interstitial PO2, produced by a twitch in a rhythmically contracting muscle. A possible mechanism is flushing capillaries with arterial blood by mechanical forces. A technical novelty is the PO2 measurement with a “stroboscopic mode” and progressively increasing delay between stimulator pulse and PO2 measuring. That permitted a 20 ms time resolution for a 205 ms spike duration, using an excitation flash rate one per second.

ATP‐Sensitive K+ Channels in Smooth Muscle Cells from Rat Mesenteric Lymph Vessels VESSELS

The ATP‐sensitive potassium (KATP) channel is expressed in plasma membranes of many types of smooth muscle cells (SMCs). It is disinhibited when intracellular ATP concentrations are reduced during conditions of metabolic stress, and the resulting K+ efflux through KATP channels mediates membrane hyperpolarization and SMC relaxation. However, in the lymphatic smooth muscle cells (LSMCs) of rhythmically contracting lymphatic vessels, KATP channels reportedly play an important role in regulating the basal rhythmic contractions that establish lymph flow even in the absence of overt metabolic challenge. The KATP channels in LSMCs also may participate in the dilator responses of lymphatic vessels (LVs) to nitric oxide, inflammatory mediators, and compromise lymph flow in other pathologies associated with oxidative stress1,2,3. Although it is recognized that the properties and the subunit composition of KATP channels show tissue‐specificity, the identity and properties of KATP channels in LSMCs are unknown. The goal of this study was to begin to define the function and properties of KATP channels in rat mesenteric LVs. Isolated rat mesenteric LVs were cannulated on glass pipettes, pressurized at 4.5–6 mmHg, and edge detection was used to continuously monitor changes in vessel diameter. Addition of the KATP channel agonist cromakalim (0.01 μM‐100 μM; half‐log units) progressively increased LV diameter and abolished rhythmic contractions (n=3). In contrast, addition of the KATP channel antagonist, glibenclamide (0.01 μM‐100 μM; half‐log units) resulted in no change in LV diameter but suppressed rhythmic contractions. The latter finding confirmed the important role of KATP channels as regulators of LV contractile function under basal conditions. Subsequently, we have begun to characterize KATP channel currents in SMCs freshly isolated from rat mesenteric LVs. Patch‐clamp studies in the cell‐attached mode reveal unitary K+ currents in symmetrical 145 K+ recording solutions at negative patch potentials near the resting membrane potential of LSMCs. Unitary amplitudes calculated over a range of negative patch potentials from −80 mV to −30 mV indicate a single‐channel conductance of ~ 37 pS. Thus, we have tentatively identified the KATP channel in rat LSMCs as a first step toward defining it's regulation by endogenous substances and pharmacological interventions that may alter lymph flow.Support or Funding InformationSupported by T32 GM106999 (BRG) and R21 CA187325‐01A1 (NJR) from the National Institutes of Health.

Comparison of Cardiomyogenic Potential among Human ESC and iPSC Lines

We recently reported that, following induction of clumps of pluripotent H1 human embryonic stem cells (hESCs) with activin-A and Bmp4 in defined medium for 5 days, widespread differentiation of rhythmically contracting cardiomyocytes occurs within 3-4 weeks. In this study, the same approach was used to assess whether human induced pluripotent stem cells (hiPSCs), which may theoretically provide an unlimited source of patient-matched cells for transplantation therapy, can similarly undergo cardiomyocyte differentiation. Differentiation of four pluripotent cell lines (H1 and H9 hESCs and C2a and C6a hiPSCs) was compared in parallel by monitoring rhythmic contraction, morphologic differentiation, and expression of cardiomyogenic genes. Based on expression of the cardiomyogenic lineage markers MESP1, ISL1, and NKX2-5, all four cell lines were induced into the cardiomyogenic lineage. However, in contrast to the widespread appearance of striations and rhythmic contractility seen in H9 and especially in H1 hESCs, both hiPSC lines exhibited poor terminal differentiation. These findings suggest that refined modes of generating hiPSCs, as well as of inducing cardiomyogenesis in them, may be required to fulfill their potential as agents of cardiac regeneration.

Role of Secretory Phospholipase A2 in Rhythmic Contraction of Pulmonary Arteries of Rats With Monocrotaline-Induced Pulmonary Arterial Hypertension

Excessive stretching of the vascular wall in accordance with pulmonary arterial hypertension (PAH) induces a variety of pathogenic cellular events in the pulmonary arteries. We previously reported that indoxam, a selective inhibitor for secretory phospholipase A2 (sPLA2), blocked the stretch-induced contraction of rabbit pulmonary arteries by inhibition of untransformed prostaglandin H2 (PGH2) production. The present study was undertaken to investigate involvement of sPLA2 and untransformed PGH2 in the enhanced contractility of pulmonary arteries of experimental PAH in rats. Among all the known isoforms of sPLA2, sPLA2-X transcript was most significantly augmented in the pulmonary arteries of rats with monocrotaline-induced pulmonary hypertension (MCT-PHR). The pulmonary arteries of MCT-PHR frequently showed two types of spontaneous contraction in response to stretch; 27% showed rhythmic contraction, which was sensitive to indoxam and SC-560 (selective COX-1 inhibitor), but less sensitive to NS-398 (selective COX-2 inhibitor); and 47% showed sustained incremental tension (tonic contraction), which was insensitive to indoxam and SC-560, but sensitive to NS-398 and was attenuated to 45% of the control. Only the rhythmically contracting pulmonary arteries of MCT-PHR produced a substantial amount of untransformed PGH2, which was abolished by indoxam. These results suggest that sPLA2-mediated PGH2 synthesis plays an important role in the rhythmic contraction of pulmonary arteries of MCT-PHR.

Collagen gels populated with rat neonatal heart cells can be used for optical recording of rhythmic contractions which also show ECG-like potentials.

Neonatal heart cells populated collagen gels show rhythmic contractions which can be recorded optically and electrically. Optical recordings revealed two populations of rhythmically contracting gels: 1) highly coherent contracting collagen gels with normally distributed contraction interval times and contraction amplitudes, and 2) irregularly contracting gels with a multi-modal distribution of contraction interval times and amplitudes. The irregularly contracting gels were shown to be 'semi regular', which means that a short contraction interval was preferentially followed by another short interval. The volume of the collagen gel during the contraction decreased, and our calculations indicate that the myocytes expel 3-10 times their own volume from the gel. Changes in electrical potential were observed depending on the location of the electrodes. These electrical, ECG-like changes in potential were maximal when one electrode was placed in the centre and the other at the edge of the gel. The results of this study indicate that myocyte-populated collagen gels are a very promising system for studies of electrophysiology and coherent contractions.

Does the pressor response to ischemic exercise improve blood flow to contracting muscles in humans?

The purpose of this study was to determine in humans 1) the gain for the reflex pressor response that occurs when perfusion pressure to rhythmically contracting muscles is reduced and 2) whether the pressor response improves blood flow to the contracting muscles. Six normal subjects performed light, moderate, and heavy rhythmic forearm contractions (30/min) with the forearm enclosed in a Plexiglas box. Pressure in the box was increased 10 mmHg each minute up to 50 mmHg to reduce transmural pressure in the arterial system of the forearm. Mean arterial pressure (MAP) was measured continuously. During light exercise no reflex increase in MAP occurred until box pressure was 50 mmHg. During moderate and heavy exercise MAP began to increase with only 10- to 20-mmHg increases in box pressure. The slope of this increase was 3.5-3.9 mmHg per 10 mmHg of box pressure (approximately 60% of that in dogs). In a further study on six subjects a deep vein draining the active forearm muscles was cannulated and deep venous O2 saturation measured to assess how a 50-mmHg increase in box pressure and subsequent reflex increase in MAP altered blood flow to the contracting muscles during heavy rhythmic exercise. The increase in box pressure reduced blood flow to contracting forearm muscles by 20-25% and was followed by a 19-mmHg increase in MAP that did not appear to improve perfusion of the active muscles. This finding was unexpected, because studies in dogs suggest that the pressor response to rhythmic exercise with restricted muscle blood flow can improve perfusion of the active muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Matching neural and muscle oscillators: control by FMRFamide-like peptides

Stomatogastric nervous systems of the shrimp, Palaemon serratus, were stained with antisera raised against the peptide FMRFamide. FMRFamide-like immunoreactivity was found in fibers in the input nerve to the stomatogastric ganglion (STG), in several STG somata, in dense neuropil in the STG, in the motor nerves that innervate the dilator muscles of the pyloric region, but not in the pyloric dilator (PD) motor neurons. FMRFamide and several FMRFamide-like peptides elicited sequences of rhythmic depolarizations and contractions of the pyloric dilator muscle. As peptide concentrations were increased, a discrete threshold for contraction was found, above which contractions were initiated with a decreasing latency in an all-or-none fashion. Muscles stopped rhythmically contracting after many seconds to several minutes of activity; the duration of spontaneous oscillatory activity in peptide was proportional to the concentration of applied peptide. In the absence of peptide, each motor neuron discharge evoked small graded muscle contractions. During peptide-induced oscillations, motor neuron activity did not always entrain muscle oscillations. After spontaneous oscillations had stopped, when the motor neurons were stimulated in the presence of the peptide, each motor neuron burst evoked large amplitude contractions as a result of the peptide-induced regenerative properties of the muscle membrane.

Heart induction in wild‐type and cardiac mutant axolotls (Ambystoma mexicanum)

We have re-examined some of the factors affecting the induction of heart-forming mesoderm in the axolotl. The formation of functional, rhythmically contracting myocardial tissue was used as an assay. We have found that heart-forming mesoderm is fully induced and capable of completing its developmental repertoire by the end of neurulation. As has been previously reported, pharyngeal endoderm appears to be the major inductor of heart mesoderm. Unlike previous workers, we have found that the inducing activity appears to be highly localized in the mid-ventral pharyngeal endoderm. The endoderm retains its inductive properties, and the mesoderm retains at least some capacity to respond, long after the heart-forming mesoderm is apparently fully induced. We have also found that RNA extracts from pharyngeal endoderm, which are capable of causing cardiac-lethal (c/c) mutant axolotl hearts to begin beating, are not capable of inducing early wild-type heart-forming mesoderm. Based on these results, we speculate that induction of heart-forming mesoderm is a two-step process. The first signal, occurring during neurulation, directs the mesoderm to begin differentiating into cardiomyocytes, and the second, beginning in mid- to late neurulation and continuing until just prior to the onset of heartbeat, causes myofibrillogenesis and the initiation of rhythmic contractions. The latter signal, which is lacking in c/c mutant embryos, appears to be necessary to override an inhibition present in the embryonic milieu.

The effects of topical capsaicin on rat urinary bladder motility in vivo

The effects of topical capsaicin on urinary bladder motility were investigated following saline-induced distension of the bladder wall in urethane-anaesthetized rats and compared to the effects of topical substance P and acetylcholine. Capsaicin and substance P produced similar excitatory effects in both quiescent and rhythmically contracting bladders, i.e., a TTX resistant tonic contraction followed by a series of rhythmic, TTX sensitive, phasic contractions. Acetylcholine, in doses equieffective in producing TTX resistant contractions was less effective than capsaicin or substance P in triggering neurogenic rhythmic contractions of bladder muscle. Atropine pretreatment prevented the neurogenic component of the excitatory effect of both capsaicin and substance P. Repeated applications of capsaicin but not of substance P led to desensitization. Bladders of animals pretreated (4 days before) with a large dose of s.c. capsaicin developed insensitivity to topical capsaicin and a larger volume of saline was required to trigger neurogenic rhythmic contractions of the detrusor muscle. These results suggest that capsaicin acts by interfering with the mechanism(s) regulating the threshold for the micturition reflex to occur.

Effect on arterial pressure of rhythmically contracting the hindlimb muscles of cats.

Although static contraction of the hindlimb muscles of anesthetized cats is known to reflexly increase arterial pressure and heart rate, the cardiovascular effects of rhythmic contractions of these muscles is unclear. To help clarify this issue, we determined, in chloralose-anesthetized cats, the effects on arterial pressure and heart rate of rhythmically contracting the hindlimb muscles at a frequency of 5 Hz. In addition, we determined the effect of rhythmic contractions on the impulse activity of group III and IV muscle afferents whose activation is known to increase cardiovascular function. We found that rhythmic contractions increased arterial pressure (from 108 +/- 8 to 134 +/- 9 mmHg; P less than 0.05) and heart rate (from 192 +/- 13 to 208 +/- 10 beats/min; P less than 0.05) in 10 cats and decreased arterial pressure (from 107 +/- 8 to 93 +/- 9 mmHg; P less than 0.05) but did not change heart rate in 9 other cats. The increases were reflex, because they were prevented by cutting the spinal roots innervating the contracting hindlimb. The decreases, however, were not reflex, because they persisted after spinal root section. The differences in the arterial pressure responses to rhythmic contractions may have been partly due to individual differences in the level of anesthesia, because in three cats the pressor responses to this maneuver were converted to depressor responses after giving the cats additional chloralose. Rhythmic contractions of the triceps surae muscles stimulated 8 of 10 group III afferents and 9 of 16 group IV afferents. We conclude that rhythmic contraction is capable of reflexly increasing cardiovascular function in cats provided that the effect is not depressed by anesthesia.

In vitro determination and differentiation of mouse teratocarcinoma toward myocardiogenesis

Pluripotential mouse teratocarcinoma cells in embryoid bodies were explanted in vitro after which various modes of differentiation occurred. After about 5 days in culture, 1–4% of the piled-up colonies exhibited synchronized rhythmical contraction with a constant frequency ranging from 50 to 70 contractions/min, irrespective of the colony size. This phenomenon could be maintained for 25 days under our culture conditions. Ultrastructural studies of the rhythmically contracting colonies revealed determination and differentiation of embryonal carcinoma cells exclusively toward myocardiogenesis, and the developmental cellular stages were observed. Myocardial differentiation was characterized by the developmental stages of fibrillogenesis, accompanied by typical specialization of the myocardial-membrane complex. This myocardial differentiation bears a close similarity to the normal embryonic myocardiogenesis of 8- to 18-day mouse embryo heart.

Blood flow through limb muscles during heavy rhythmic exercise.

AbstractThe effect of rhythmically performed muscle contractions on blood flow through muscles was investigated in the calf of cat. During a series of contractions, which had led to maximal dilatation of the resistance vessels, a considerable lowering of the pressure in a small vein from the contracting muscles could be demonstrated between contractions, despite the huge inflow. An optimal effect was obtained when the muscle nerve was stimulated with a short (200–300 msec) impulse train once per sec, a pattern mimicking that during running. Rhythmic contractions of this type led to a characteristic blood flow pattern through the muscles. Arterial inflow to the muscle vascular bed occurred only in between contractions, whereas nearly all the venous outflow from the muscles occurred during the contractions.The venous pressure reduction occurring between contractions implies a gain in effective perfusion pressure and a corresponding augmentation in blood flow through the muscles. Such an effect was found to be considerable in dependent limbs. Here average blood flow through the rhythmically contracting calf muscles could be as great as, or even greater than, the “free” flow in the immediate post‐exercise period. This effect of the “muscle pump” may be of great importance also in the dependent limbs of man.

Specificity of the effects of cardiac glycosides on the rhythmic contraction of single cultured cardiac muscle cells.

FROM fragments of embryonic chick hearts treated with trypsin, single, rhythmically contracting cells can be grown in a fluid, semisynthetic culture medium1. Exposure of these cells to cardiac glycosides in concentrations as low as 10−9–10−8 M results in an increase in the rate of beat2. In addition the pulsations appear to become stronger. At concentrations of 10−7 M and above, the acceleration of pulsation is soon followed, or is accompanied, by disturbances of automaticity and of co-ordinated contractile activity. By means of phase-contrast, high-speed microcinematography of cells anchored to the glass surface these effects could be studied in greater detail3. The motion pictures revealed changes at the cellular level which strikingly resembled some of the familiar manifestations of digitalis poisoning in the heart of animals and of man. Such phenomena as partial and complete block of propagation of the contraction wave (presumably due to block of conduction), bigeminal rhythm, ectopic beats, tachycardia, and fibrillation could all be observed in single isolated ventricle cells.

THE EFFECT OF CERTAIN CALCIUM SALTS ON THE RHYTHMICALLY CONTRACTING AND QUIESCENT UTERINE FISTULA, WITH OBSERVATIONS ON THE ACTION OF POSTERIOR PITUITARY EXTRACTS

ArticleTHE EFFECT OF CERTAIN CALCIUM SALTS ON THE RHYTHMICALLY CONTRACTING AND QUIESCENT UTERINE FISTULA, WITH OBSERVATIONS ON THE ACTION OF POSTERIOR PITUITARY EXTRACTSSamuel R. M. ReynoldsSamuel R. M. ReynoldsFrom the Department of Physiology, School of Medicine, Western Reserve University, Cleveland, OhioPublished Online:31 Jul 1933https://doi.org/10.1152/ajplegacy.1933.105.2.358MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation More from this issue > Volume 105Issue 2July 1933Pages 358-365 Copyright & PermissionsCopyright © 1933 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1933.105.2.358History Received 14 May 1933 Published online 31 July 1933 Published in print 31 July 1933 Metrics