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Endurance exercise with and without a thermal clamp: effects on leukocytes and leukocyte subsets.

To test how leukocyte responses to endurance exercise were modified by clamping body temperature, nine men (27.3 +/- 6.0 yr) completed four 80-min immersions to midchest at water temperatures of 23 or 39 degrees C; two tests included 40-min of cycle ergometer exercise at 65% of aerobic power. When the subjects were exercising, rectal temperature peaked at 39.1 +/- 0.4 degrees C in the warm water and 37.8 +/- 0.3 degrees C in the cool water. When the subjects were sitting in warm water, rectal temperature closely matched the core temperature during exercise in cool water, whereas when they were sitting in cool water, rectal temperatures decreased to 36.4 +/- 0.6 degrees C. Total and differential white cell counts were determined by using a Coulter counter, and cortisol and growth hormone concentrations were determined by radioimmunoassay; all data were adjusted for changes of blood and plasma volumes. Heat clamping during exercise substantially reduced the rise in white cell, lymphocyte, and granulocyte counts but not the increase in monocyte count. Clamping also abolished previously observed associations between cell counts and cortisol and weakened associations with growth hormone concentrations (D. A. McCarthy and M. M. Dale. Sports Med. 6: 333-363, 1988). We conclude that both exercise and a rise of core temperature contribute to the changes in white cell and subset counts during and immediately after moderate exercise. Both cortisol and growth hormone concentrations appear to be mediators of these responses.

Adult↗

Voltage-clamp analysis of a Ca2+- and voltage-dependent chloride conductance in cultured mouse spinal neurons.

Current and voltage-clamp recordings were made at room temperature from cultured mouse spinal neurons using conventional two-electrode voltage-clamp techniques and electrodes filled with either 3 M KCl, 3 M CsCl, or 3 M Cs2SO4. In the presence of tetraethylammonium and tetrodotoxin, "fast" (rapidly rising and falling) action potentials (FAP) of variable duration were recorded in most neurons. "Slow" (slowly rising and falling) depolarizing potentials (SDP) occurred in 23% of the cells, when using KCl-filled electrodes, and in 82% of the cells with CsCl-filled electrodes. The SDP was frequently preceded by an FAP, although in some cells activation of the SDP occurred before the FAP threshold was reached and in a graded fashion. Both the FAP and SDP were abolished by Cd2+ and other Ca2+ antagonists. In cells exhibiting SDPs, voltage-clamp analysis revealed a sustained (noninactivating) inward current (Isin) during depolarizing steps to potentials more positive than -45 mV. Repolarizing steps resulted in slowly decaying inward tail currents (Itail). Both Isin and Itail were abolished in solutions nominally free of Cao2+, or containing Ca2+-channel antagonists. Bao2+ did not support Isin. The data indicated a U-shaped activation curve for Isin, peaking at about -10 mV. Activation of Isin occurred exponentially with a time constant of approximately 140 ms at -23 mV, becoming faster at more depolarized potentials (ca. 50 ms at -2 mV). Deactivation was slow, giving rise to tail currents lasting seconds. In some cases deactivation could be described by a single exponential process, although frequently the kinetics were more complex. Deactivation was faster at hyperpolarized potentials and sensitive to extracellular ([Ca2+]o), duration of activating voltage steps, and the degree of activation of Isin. Using CsCl-filled electrodes, the reversal potential (Erev) for Isin was -1.7 mV (SEM 3.5 mV, n = 20). Erev always corresponded to the reversal potential for gamma-aminobutyric acid-evoked currents in the same cell. In experiments in which Cs2SO4-filled electrodes were used, Erev was estimated to be -44 mV (SEM 2.3 mV, n = 9). Neither complete substitution of Nao+ with choline ions nor elevation of [K+]o 10-fold significantly affected the estimated Erev. However, substitution of Cl0- with isethionate or methanesulphonate increased the amplitude of inward currents (recorded with CsCl-filled electrodes) and shifted Erev to more depolarized potentials. The results indicate that Cl- are the primary charge carriers for this current and that Cai2+ is required for its activation, leading us to identify it as ICl(Ca).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Unitary conductance changes at teleost Mauthner cell glycinergic synapses: a voltage-clamp and pharmacologic analysis.

1. The magnitude and kinetics of inhibitory postsynaptic currents (IPSCs) evoked in the goldfish Mauthner (M-) cell by intracellular stimulation of identified presynaptic interneurons (unitary responses) and by activation of the recurrent collateral network were determined with single-and double electrode voltage-clamp techniques. 2. The peak magnitude of the inhibitory conductance changes were 5610 +/- 4800 nS (mean +/- SD; n = 13) for the collateral response, and 144 +/- 44 nS (n = 7) for the unitary IPSCs. These synaptic conductances, which are due to the opening of Cl- channels, were independent of the degree of Cl- -loading of the M-cell. 3. The peak amplitude of the collateral inhibitory postsynaptic potential (IPSP) was a constant fraction (0.52 +/- 0.06) of the driving force, which was determined from current-voltage plots for both types of IPSCs and ranged from 10 to 37 mV. These findings confirm indirect measurements from previous current-clamp studies and validate the normalization procedure used to previously calculate synaptic conductances from IPSP amplitudes, a method that therefore may be applicable to other central neurons. 4. At the resting membrane potential, the rise time of the unitary IPSCs was 0.34 +/- 0.07 ms (n = 18), whereas their decay was exponential, with a time constant of 5.7 +/- 1.1 ms (n = 16). 5. Iontophoretic and intramuscular applications of the glycine antagonist strychnine reduced or blocked M-cell inhibitory responses, without altering the excitability of the presynaptic neurons, or the driving force. 6. Amplitude fluctuations of unitary IPSPs recorded during partial blockade by strychnine were analyzed according to a binomial model of quantal transmitter release. In one experimental series, comparison of the binomial parameters before and after applying the antagonist indicated that only quantal size, q, was reduced, whereas n, the number of available release units, and p, the probability of release, were unaffected by strychnine. In a second series, the individual presynaptic cells were injected with horseradish peroxidase (HRP), and it was found that the correlation between n and the number of stained presynaptic boutons and, therefore, of active zones, was maintained in the presence of the drug. No evidence was found for silent synapses in these conditions. 7. The quantal conductance, gq, was estimated from the binomially derived quantal size, in millivolts, and the voltage-clamp measurements of the IPSP driving force and M-cell input conductance. gq averaged 21.5 nS in control conditions and 12.3 nS in the presence of strychnine.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Single-electrode voltage-clamp analysis of the N-methyl-D-aspartate component of synaptic responses in neocortical slices from children with intractable epilepsy.

1. Synaptic transmission mediated by the N-methyl-D-aspartate (NMDA)-receptor type was studied in neocortex from children undergoing surgical treatment for intractable epilepsy. Intracellular recordings from pyramidal cells were obtained in slices of neocortical tissue by use of microelectrodes. Synaptic responses were induced by electrical stimulation and studied with current-clamp and single-electrode voltage-clamp techniques. The NMDA-receptor-mediated component of the synaptic responses was isolated by addition of 10 microM bicuculline and 30 microM 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX) in the perfusion solution. 2. In the presence of bicuculline and CNQX, electrical stimulation evoked an excitatory postsynaptic potential (EPSP) in every recorded cell. The amplitude of this EPSP increased when membrane potential was depolarized with injected current. 3. All cells studied in voltage clamp were recorded with microelectrodes containing Cs+ and QX 314. To avoid contamination of the responses from voltage-dependent Ca2+ conductances, membrane potential was held at depolarized potentials until Ca2+ spiking inactivated completely. The evoked excitatory postsynaptic currents (EPSCs) measured at resting membrane potential ranged from 100 to 400 pA. The NMDA receptor-selective antagonist DL-2-amino-5-phosphonopentanoic acid (AP-5) reversibly decreased the current amplitude by 60% for 10 microM and 80% for 30 microM. 4. The current-voltage (I-V) relation showed a region of negative slope conductance between -100 and -20 mV. The largest currents (-250 to -900 pA) were recorded in the range of -45 to -20 mV and reversed between -10 and +10 mV. Removing Mg2+ from the perfusion solution decreased the negativity of the slope, which is consistent with a reduction in the voltage-dependent Mg2+ block of the NMDA-receptor channel. 5. The I-V plots obtained from cells recorded in the most abnormal tissue were averaged and compared with those from the least abnormal tissue. No significant difference was found between these two groups. The averaged plots from the youngest patients (8 and 10 mo old) and those from the oldest (5-15 yr old) patients were also compared, and the results from these two groups were not significantly different.(ABSTRACT TRUNCATED AT 400 WORDS)

2-Amino-5-phosphonovalerate↗

Dynamic clamp: computer-generated conductances in real neurons.

1. We describe a new method, the dynamic clamp, that uses a computer as an interactive tool to introduce simulated voltage and ligand mediated conductances into real neurons. 2. We simulate a gamma-aminobutyric acid (GABA) response of a cultured stomatogastric ganglion neuron to illustrate that the dynamic clamp effectively introduces a conductance into the target neuron. 3. To demonstrate an artificial voltage-dependent conductance, we simulate the action of a voltage-dependent proctolin response on a neuron in the intact stomatogastric ganglion. We show that shifts in the activation curve and the maximal conductance of the response produce different effects on the target neuron. 4. The dynamic clamp is used to construct reciprocal inhibitory synapses between two stomatogastric ganglion neurons that are not coupled naturally, illustrating that this method can be used to form new networks at will.

Animals↗

Dependence of LTP induction on postsynaptic depolarization: a perforated patch-clamp study in visual cortical slices of young rats.

1. To see whether there is a threshold of postsynaptic depolarization for induction of long-term potentiation (LTP) or depression (LTD) of synaptic transmission, perforated patch-clamp recordings were carried out under microscopic observation from 61 layer II/III neurons in visual cortical slices of young rats. Electrical stimulation given to nearby neurons was paired with stepwise shifts (30- or 300-ms duration) of clamped membrane potential of the recorded neurons to various levels. 2. Excitatory postsynaptic currents (EPSCs) were elicited by focal stimulation of a nearby pyramidal cell-like neuron. As the intensity of stimulation was increased, EPSCs emerged abruptly with 100% probability, and their peak latencies and amplitudes remained almost constant up to more than twice the threshold, indicating that the EPSCs were elicited monosynaptically. 3. LTP of EPSCs was induced in 11 of the 15 cells after pairing with a step to -20 mV and in 5 of the 14 cells after pairing with a step to -40 mV. No LTP was observed when the postsynaptic cells were clamped at -60, -70, or -90 mV. Significant LTD was not seen at any membrane potential level tested. There was no significant difference between the duration of potential shift of 30 and 300 ms during the pairing procedure in induction probability of LTP and magnitude of LTP, if it was induced. 4. These results suggest that LTP is induced by synaptic inputs associated with postsynaptic depolarization above the threshold around -40 mV at synapses linking layer II/III neurons in the developing visual cortex.

Age Factors↗

Distribution and functional properties of 5-HT3 receptors in the rat hippocampal dentate gyrus: a patch-clamp study.

1. In dentate gyrus of rat hippocampal slices two distinct types of neurons, principal excitatory neurons (granule cells) and local inhibitory neurons (basket cells), could be identified under Nomarski microscopy; I investigated the actions of serotonin using the whole-cell patch-clamp technique. The identification of the neurons was later confirmed by intracellular staining with Lucifer yellow. 2. In both basket cells and granule cells, whole-cell current recordings revealed spontaneous synaptic currents ranging from < 10 pA to > 200 pA in symmetrical Cl- conditions at a holding potential of -63 mV. These currents were blocked by 10 microM bicuculline, indicating that they resulted from the spontaneous activation of GABAergic inputs (which had been morphologically described in both types of neurons). 3. By focal application of serotonin (2-50 microM) to basket cells under current clamp I evoked a train of action potentials superimposed on a baseline membrane depolarization. Under voltage-clamp conditions serotonin evoked an inward current at a holding potential of -63 mV (currents were detectable in approximately 90% of basket cells studied). The inward current was accompanied by a multitude of small inward currents of short duration (< 100 ms) that were found to be due to the stimulation by serotonin of nearby GABAergic presynaptic neurons innervating the recorded neuron. 4. In granule cells (total of 11 cells) serotonin did not produce any responses under conditions similar to those used for basket cells. The occurrence of bicuculline-sensitive spontaneous synaptic current events seemed to increase during the application of serotonin; this phenomenon reflected the excitatory action of serotonin exclusively on GABAergic interneurons. 5. The serotonin-induced inward currents in basket cells were mediated by the 5-HT3 receptor subtype because 1) they were blocked by either metoclopramide (10 microM) or [3-alpha-tropanyl]-1H-indolecarboxylic acid ester (2 nM), the latter being a specific blocker for the 5-HT3 receptor subtype, and 2) almost similar currents were induced by the application of the selective 5-HT3 receptor agonist 2-methyl 5-HT (2-50 microM) or 1-(m-chlorophenyl)-biguanide (0.1-10 microM). 6. Current-voltage (I-V) relations of serotonin-induced currents in basket cells showed that the reversal potential was close to 0 mV in external standard saline and depended on the concentrations of monovalent cations. I-V relations of serotonin-induced currents revealed inward rectification at the membrane potential range of +30 to -60 mV.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

'On-line' alterations of contralateral jugular blood gas profile during carotid clamping.

The aim of this work was to record the metabolic status of the brain (pH, PCO(2) and PO(2)) during carotid endarterectomy (CEA), with the use of an intravascular multiparameter sensor (Paratrend 7) via retrograde catheterization of the contralateral jugular vein. Twenty-four patients with ASA grades II and III scheduled for CEA were included in the study. After induction of anesthesia, the contralateral internal jugular vein was punctured retrogradely and the sensor was introduced. During clamping, pH became persistently more acidotic (7.34-7.31; p < 0.05), PCO(2) was elevated (43.2-46.8 mm Hg; p < 0.05) while most of the patients showed a non-significant decrease in PjvO(2)/SjvO(2) (jv = jugular venous). Correlation with clamping time or stump pressure was not significant. Unclamping was followed by a short period (5- 9 min) of decrease in pH and elevation of PCO(2) (7.30-7.22; p < 0.05, and 48.0-52.5 mm Hg; p < 0.05, respectively). PjvO(2) was significantly elevated (51.8-58.0 mm Hg; p < 0.001) after the restoration of flow. The study suggests that local CO(2) creates conditions for compensation of flow after the application of a carotid clamp. We consider that this monitoring technique, after further validation, may provide useful information.

Aged↗

Changes in hemodynamics and acid-base balance during cross-clamping of the descending thoracic aorta. A study in patients operated on for thoracic and thoracoabdominal aortic aneurysm.

AIM OF THE STUDY: In the clinical situation there is discrepancy between various investigations regarding the cardiac response of thoracic aortic cross-clamping. The aim was therefore to investigate the hemodynamic response and blood gases during proximal aortic cross-clamping (XC) in patients operated for descending thoracic and thoracoabdominal aortic aneurysm without circulatory support. PATIENTS AND METHODS: Altogether 51 patients operated on for thoracoabdominal (n=31) or descending thoracic aortic aneurysm (n=20) were included in the investigation. All patients were operated with aortic XC, but no circulatory support was applied. Hemodynamic variables and blood gases were recorded before and during XC. RESULTS: A significant increase in cardiac output during XC from 4.7 to 6.0 liters/min was observed (p<0.01). There was a similar percentual increase in heart rate and also the proximal systolic blood pressure increased. A metabolic acidosis occurred during XC. CONCLUSION: Cardiac output was significantly increased during XC in patients operated on for thoracoabdominal or descending thoracic aneurysm using direct aortic XC without circulatory support. Simultaneously, the heart rate was increased and there was a hyperdynamic circulatory state proximal to the aortic clamp. Redistribution of the blood volume in addition to catecholamine release may be responsible for the observed changes. These observations may influence the selection of operative strategy for some of these patients.

Acid-Base Equilibrium↗

Look twice before you clamp: decapitation of an omphaloenteric duct. A case report.

OBJECTIVE: To report a case of accidental decapitation of a persistent omphaloenteric duct (POD) in congenital hernia of the umbilical cord in a newborn male. CASE PRESENTATION AND INTERVENTION: The newborn was transferred to an intensive care unit, where under closer inspection the diagnosis of an omphalocele was presumed. Surgical exploration at our Pediatric Surgical University Clinic revealed a clamped and decapitated POD in the umbilical cord. The decapitation had gone unnoticed due to severe asphyxia resulting from meconium aspiration encountered at the time of delivery. Using general anesthesia, the clamp was removed under sterile conditions. The decapitation of the POD in the herniated umbilical cord was confirmed at this time. The severed intestine was sutured and closed using 5-0 Vicryl sutures. The further course was uneventful. CONCLUSION: Herniation of the umbilical cord at the time of birth by intestinal loops or POD is rare, but caution should be taken by midwives before clamping in order to avoid the type of injury seen in this case.

Hernia, Umbilical↗

A new head clamp for stereotactic and intracranial procedures. Technical note.

The authors have devised a clamp to fix the head to the surgical table during stereotactic procedures for placement of depth electrodes. The main advantages of this device derive from the rigid immobilization of the head. The clamp facilitates fixation of the stereotactic frame to the head, percutaneous twist drill trephination of any site of the skull, X-ray centering with a laser beam, mobilization of contrast material during ventriculography or pneumoencephalography, and placement of epidural screws. This head clamp can also be used during craniotomies for fixation of the head and support for brain retractors.

Brain Diseases↗

A biochemical and ultrastructural study on myocardial changes during aorto-coronary bypass surgery: St. Thomas Hospital cardioplegia versus intermittent aortic cross-clamping at 34 and 25 degrees C.

The changes induced by continuous aortic cross-clamping in combination with multidose ice-cold St. Thomas Hospital cardioplegia (myocardial temperature below 16 degrees C), or intermittent aortic cross-clamping at 34 or 25 degrees C were evaluated in a randomized study on 72 patients undergoing extensive aorto-coronary bypass surgery. The cumulative release of heart-specific enzymes was very small and no marked ultrastructural changes in mitochondria of both the subepi- and the subendocardial layer of the left ventricular free wall occurred. No differences between the three operation techniques could be observed on the basis of the above-mentioned parameters. Myocardial ATP and glycogen contents were decreased in post-ischaemic tissue in both the normothermic and hypothermic intermittent aortic cross-clamp groups. This decrease was associated with a release of lactate and inorganic phosphate during the repetitive periods of reperfusion. No change in myocardial ATP and glycogen content could be observed in the cardioplegia-treated hearts. St. Thomas Hospital cardioplegia is obviously most effective in preventing changes in myocardial metabolism such as reduction of ATP and carbohydrate stores during the reversible phase of ischaemic insult.

Adenosine Triphosphate↗

Maximal upstroke velocity as an index of available sodium conductance. Comparison of maximal upstroke velocity and voltage clamp measurements of sodium current in rabbit Purkinje fibers.

We compared the maximal upstroke velocity of action potentials in short rabbit Purkinje fibers with sodium currents measured with a two-microelectrode voltage clamp. The number of sodium channels available to open during a sudden depolarization was varied either by blockade with tetrodotoxin or by inactivation with steady depolarizations. In both cases, the maximal upstroke velocity was found to be a very nonlinear measure of the number of available sodium channels. For example, 3 microM tetrodotoxin blocks 85% of the sodium channels, but reduces the maximal upstroke velocity by only 33%. Voltage clamp and upstroke velocity experiments were reconstructed with a computer model of the rabbit Purkinje fiber preparation that was closely based on experimental measurements of passive cable properties and sodium channel characteristics. The simulations indicate that our voltage clamp measurements of sodium current accurately report changes in channel availability, but they also show that the maximal upstroke velocity is a strongly nonlinear index of available sodium conductance. Most of the nonlinearity arises from the activation kinetics of the sodium channels: as the pool of available channels decreases, a greater percentage of those channels activate and contribute inward current at the time of the maximal upstroke velocity. Simulations predict that the maximal upstroke velocity-available sodium conductance relationship would still remain nonlinear at 37 degrees C or under different stimulus conditions that give uniform or continuously propagated action potentials. The nonlinearity may invalidate inferences based on earlier maximal upstroke velocity experiments: the existence of two types of sodium channels with different tetrodotoxin sensitivity, steady state voltage dependence of tetrodotoxin block, voltage range over which sodium channels inactivate, and rapid, then slow recovery of sodium channel availability following a sudden repolarization. All of these conclusions need to be reevaluated.

Action Potentials↗

Left ventricular internal resistance and unloaded ejection flow assessed from pressure-flow relations: a flow-clamp study on isolated rabbit hearts.

Left ventricular pressure-flow relations were studied, using excised working rabbit hearts and imposing constant flow ejections (flow-clamps) to separate the effects of flow on pressure from those of time, flow duration, starting volume, ejected volume, and volume at specified time. Pressure-flow data at given volume and time were independent of flow duration, starting volume, and ejected volume for flow-clamp durations exceeding 30 msec. Flow history independent of pressure-flow relations was linear for flow values larger than +/- 5 ml/sec. The time-varying elastance model, E(t), of the ventricle was extended with a resistive component. Transient effects of flow can be explained by including a second elastance. The resulting verified 3-component model is consistent with recent reported experimental findings. The properties of internal resistance correspond to a constant unloaded ejection flow Qmax, which was tested by extrapolating the linear pressure-flow relations to zero pressure. Qmax reached a plateau value of approximately 25 ml/sec within 50 msec after the start of contraction. In relaxation, Qmax is only slightly smaller. Qmax did not depend on volume; therefore, the following equation was adequate for the relation between pressure, p(t); volume, V(t); and flow Q(t), during the flow-clamped ejections from 30 minutes after the start of the flow: (t) = E(t).(V(t)-Vd).(1-Q(t)/Qmax)

Animals↗

Pyelorenal backflow in rabbits following clamping of the renal vein and artery: radiologic and microscopic investigation.

Retrograde pyelography was carried out in rabbits during and following temporary clamping of either the renal artery and vein together or the main renal vein alone. Pyelosinous backflow was observed in all kidneys and pyelovenous backflow in 16 of the 18 kidneys, occurring at intrapelvic pressures between 50 and 100 mmHg. During simultaneous complete renal artery and vein occlusion intrarenal backflow occurred in all 3 kidneys examined while it was observed in 3 of 6 kidneys when the pyelography was performed 5 min after removal of the clamp. During complete renal vein occlusion filling with contrast medium of the central intrarenal veins occurred in all 3 kidneys but only in 3 of 6 kidneys examined after the clamp had been removed. Microscopically, pyelosinous backflow was explained by vital tears in the fornix of the pelvic cavity. Tears extending into the kidney parenchyma were only observed in kidneys with intrarenal backflow at pyelography.

Animals↗

In situ cooling in a lung hilar clamping model of ischemia-reperfusion injury.

Experimental models for studying transplantation have up to now been unable to isolate reperfusion injury with minimal surgical manipulation and without the interference of graft rejection. Six pigs were subjected to left hilum preparation only (control group), and eight pigs were subjected to left hilum preparation plus in situ cooling ischemia and reperfusion of the lung (experimental group). The hilum was dissected free from other tissues in both groups. Lung preservation was achieved by antegrade flush perfusion via the left pulmonary artery. Pulmonary veins were clamped at the left atrium and a vent was created. The left main bronchus was clamped. Lung temperature was maintained at 4 degrees -8 degrees C, while core temperature was kept at 38 degrees C. After 3 hrs of cold ischemia the clamps were removed and the lung was reperfused. Elevated pulmonary vascular resistance and local and systemic aspects of ischemia-reperfusion syndrome were consistently reproduced. This large-animal model of in situ unilateral lung cold ischemia with warm reperfusion proved to be very reliable in reproducing all aspects of ischemia-reperfusion injury. It excludes the interference of rejection and extensive surgical manipulation. We therefore propose its use in experimental studies investigating pharmaceutical or cooling modifications affecting lung ischemia-reperfusion outcomes.

Animals↗

Excitation-contraction coupling in developing mammalian myocardium: evidence from voltage clamp studies.

The single sucrose gap voltage clamp technique was used to study excitation-contraction coupling processes in right ventricular papillary muscles from New Zealand White rabbits at various stages of development. In response to voltage clamp controlled depolarizations, muscles from newborn rabbits were found to exhibit a monotonically increasing tension response reaching a steady state level that was maintained for the duration of depolarization. In contrast, more mature myocardium responded to similar depolarizations by developing an early peak of tension before relaxing to a steady state level. Measurement of the ratio of early peak or phasic tension to steady state or tonic tension revealed a statistically significant increase in the phasic tension component with maturation. In addition, Ca2+ loading of immature myocytes via a conditioning voltage clamp step resulted in enhancement of phasic tension in subsequent test depolarizations. Finally, the voltage dependence of tonic tension was found to be the same in all age groups. In contrast, the voltage dependence of phasic tension, seen only in the more mature myocardium, differed from that of tonic tension. The results of this investigation suggest that tension development in the immature myocardium is supported largely by the influx of Ca2+ across the sarcolemma. As the myocardium matures, intracellular Ca2+ uptake and rerelease by the sarcoplasmic reticulum plays an increasingly important role in tension development. A developmental schema is presented to account for the observed maturational changes in excitation-contraction coupling.

Age Factors↗

Hyperinsulinemia suppresses glucose utilization in specific brain regions: in vivo studies using the euglycemic clamp in the rat.

It has been suggested that insulin acts centrally by altering brain glucose uptake. Previous studies of the effect of insulin on brain glucose metabolism have been difficult to interpret due to lack of steady state conditions for glucose and/or insulin. To determine whether insulin per se alters brain glucose metabolism, we measured glucose utilization rates, using the deoxyglucose method, in the medial basal hypothalamus, locus coeruleus, and motor cortex of conscious, unrestrained rats undergoing 2-h euglycemic clamps (blood glucose, 4.1 +/- 0.1 mmol/liter) at increasing insulin infusion rates. Plateau insulin levels were 29 +/- 5 mU/liter (controls) and 48 +/- 4, 146 +/- 8, 670 +/- 40, and 7560 +/- 410 mU/liter (clamped). Glucose utilization rates in the medial basal hypothalamus fell significantly from 60 +/- 4 mumol/100 g X min (controls) to 46 +/- 3, 39 +/- 2, 35 +/- 2, and 39 +/- 3 mumol/100 g X min in respective insulin-infused animals (P less than 0.01 vs. controls). Similar falls of up to 39% and 48% were seen in the locus coeruleus and motor cortex, respectively. A significant inverse correlation was found between the glucose utilization rate in each brain region and the log plasma insulin level. The reduction in glucose utilization rate was associated with marked increases in serum corticosterone levels (995 +/- 157 vs. 91 +/- 31 nmol/liter in controls, P less than 0.001). Serum potassium was significantly lower in clamped animals (5.2 +/- 0.3 to 5.9 +/- 0.3 mmol/liter) than in controls (7.0 +/- 0.4 mmol/liter, P less than 0.01). However, the inverse correlation between regional brain glucose utilization and log plasma insulin was independent of changes in serum potassium, while there was no independent correlation with serum potassium. The data reveal reduced glucose utilization in specific brain regions in the presence of insulin levels both equal to and above those found in the postabsorptive state and support a direct effect of insulin in suppressing regional brain glucose utilization.

Animals↗