The neuroendocrine response to surgical trauma is a complex set of hormonal and metabolic changes evoked by anxiety, blood loss, tissue damage, visceral handling, and also by the anaesthetic drugs and procedures (Traynor & Hall, 1981). A useful approach to understanding the surgery-induced stress syndrome is to individualize these stressful stimuli in order to explore the specific effects of anxiety, anaesthesia or surgery itself on the endocrine and metabolic responses that are observed altogether. The relative importance of each element contributing to surgical stress should be better characterized in order to provide the rationale for improving the management of surgical patients. For instance, it remains unclear whether the physiological alterations induced by surgical stress are a single syndrome arising from the addition of several stressors or comprise parallel responses to each stressor, with some convergent end-points.

In patients following myocardial infarction (MI), exercise tolerance and muscular strength are often markedly reduced. The purpose of this investigation was to determine if calcium (Ca2+) uptake and release by skeletal muscle sarcoplasmic reticulum (SR) are altered following MI and if such changes are associated with diminished muscular performance. SR vesicles were isolated from rat gastrocnemius muscle at 6 and 12 weeks following MI (via left coronary artery ligation) or sham surgery. At both post-surgery intervals, the rates of Ca2+ uptake (measured using fura-2) were 35 % greater in the MI group. In addition, the rates of Ca2+ release were increased in the MI group by 10 and 30 % at 6 and 12 weeks, respectively. At 12 weeks post-surgery, animals of the MI group showed significant reductions in in situ twitch and tetanic forces and significant elevations in the rates of tension increase and decrease. These data indicate that SR Ca2+ exchange is altered following MI. In addition, changes in SR function are associated with changes in force production by the whole muscle.

Xenopus laevis oocytes were used to analyse the effects of serotonin (5-hydroxytryptamine, 5-HT) and serotonergic agents on ionic currents elicited by the activation of mammalian muscle nicotinic acetylcholine receptors (AChRs). 5-HT as well as other serotonergic agents, such as ketanserin, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), methysergide, spiperone, or fluoxetine alone (up to 1 mM), did not elicit membrane currents in Xenopus oocytes expressing AChRs, but they reversibly reduced the current elicited by acetylcholine (ACh-current). Serotonin was applied before, together with or after ACh application, and its effects were examined on desensitizing and non-desensitizing ACh-currents. 5-HT reduced the amplitude and accelerated the desensitization of the desensitizing currents. In contrast, non-desensitizing currents were reduced in amplitude but their time course was not significantly affected. With the same concentration of 5-HT the inhibition was stronger on desensitizing than on non-desensitizing ACh-currents. For example, 100 µM 5-HT reduced the peak of a desensitizing ACh-current to 0·48 ± 0·06 (peak current ratio) and after 40 s the current was reduced to a ratio of 0·25 ± 0·04, whereas a non-desensitizing ACh-current was reduced to a ratio of 0·66 ± 0·01. All the serotonergic agents tested inhibited the ACh-currents rapidly and reversibly, suggesting that they are acting directly on the AChRs. The half-inhibitory concentration, IC50, of 5-HT acting on non-desensitizing currents elicited by 250 nM ACh was 247 ± 26 µM and the Hill coefficient was [sim]0.88, suggesting a single site for the interaction of 5-HT with the receptor. It appears that 5-HT inhibits AChRs non-competitively because neither the half-effective concentration of ACh, EC50, for ACh-current nor the Hill coefficient were affected by 5-HT. Furthermore, the extent of inhibition of 5-HT on AChRs did not depend on the nicotinic agonist (suberyldicholine, ACh or nicotine). The inhibition of AChRs by serotonergic agents was voltage-dependent. The electrical distance of the binding site for 5-HT was [sim]0.75, whereas for the other serotonergic agents tested it was [sim]0.22, suggesting that ketanserin, 8-OH-DPAT, methysergide, spiperone and fluoxetine act within the ion channel, but at a site more external than that for 5-HT. These substances inhibited the ACh-current more potently than 5-HT. We conclude that 5-HT and serotonergic agents inhibit, in a non-competitive manner, the ACh-current in muscle AChRs by blocking the open receptor-channel complex. Moreover, 5-HT appears to promote the desensitized state of the receptor when the current is elicited by high ACh concentrations.

This project aimed to quantify the regional distribution of sweat composition over the skin surface and to determine whether sweat constituent concentrations collected from regional sites can estimate whole-body concentrations. Ten males cycled for 90 min in a 20°C (50 % relative humidity) environment at 45 % peak aerobic power. Sweat was collected from eleven skin regions and the whole body, using a wash-down technique. Strong relationships were evident between the regional and whole-body sweat [Na+] and [Cl-], such that the thigh and calf exhibited greater correlation coefficients than area-weighted means derived from four and eight skin regions. Therefore, in this particular protocol the whole-body sweat [Na+] and [Cl-] could be predicted from regional sweat collections. Relationships between sweat constituents were evident for sweat [Na+] and pH, and sweat [K+] and [lactate] when data were pooled between skin regions and subjects. To our knowledge this is the first investigation to report a positive relationship between sweat [K+] and [lactate]. The exact mechanism responsible for the positive relationship between sweat [K+] and [lactate] is uncertain although it is speculated to occur at the secretory coil.

Many recent findings suggest that the nervous system has efferent effects on bone. A putative role of the sensory innervation has been assessed by using a synchronised rat model of bone resorption after treating adult animals with the neurotoxin capsaicin. Fourteen days after capsaicin treatment (50 mg kg-1) the right maxillary molars were extracted to activate a wave of resorption along the mandibular cortex. The rats were killed 4 days later (i.e. at the peak of resorption in this model), and their right mandibles were processed for histometric evaluation of resorption along the cortex and of calcitonin gene-related peptide (CGRP)- and substance P (SP)-immunoreactive (IR) fibres in the dental pulp. CGRP-IR and SP-IR fibres were significantly reduced in numbers by the capsaicin treatment (by 58 and 49 %, respectively), confirming the success of sensory denervation. The resorption surface was significantly reduced (P < 0.005) versus the sham-treated animals. Although the size of the osteoclast population recruited in the site was not modified, the number of actively resorbing osteoclasts was significantly reduced (P < 0.03). However, the activity of the resorbing cells was not modified. Non-specific esterase-positive osteoclast precursors were also significantly few after capsaicin treatment. These data show that the sensory nervous system is involved in the control of bone resorption at two different levels: (1) in the recruitment of osteoclast precursors, and (2) in regulating the access of recruited cells to the bone surface.

Spermine is a low molecular weight polyamine involved in the postnatal maturation of the gut. When it is administered orally to suckling rats, it induces maturation of the intestinal tract (liver, pancreas and small intestine). Here we show that this polyamine is able to induce precocious intestinal and splenic development in suckling mice. In fact, in 15-day-old mice which had received spermine orally twice daily for 3 days we observed an increase in the ratio of white pulp surface to total spleen surface in comparison with untreated mice. The two macrophage subsets of the marginal zone and the B-cell population were more developed and reached the development level of 5- or 10-week-old mice. The proliferation rate of B-cells was increased by spermine administration to pups. These observations suggest that spermine might play a role in immune system development; further investigation of its effects are intended, namely the evaluation of its capacity to enhance defence during the neonatal period. Experimental Physiology (2002) 87.1, 69-75.

The effects of streptozotocin (STZ) diabetes and the antihyperglycaemic agent metformin on the contractile characteristics of the limb skeletal muscles and on erythrocyte volume were examined in rats. After 8 weeks of diabetes, the tetanic tension of the extensor digitorum longus (EDL) muscle decreased and the half-relaxation time of the soleus muscle increased. Endurance decreased in both muscles. Metformin treatment of the diabetic rats did not prevent the development of these contractile changes. Diabetes induced depolarisation in the EDL and soleus muscles. Following exposure to insulin, both muscles repolarised. Metformin treatment of control rats induced depolarisation in the EDL and soleus muscles, but in the depolarised EDL and soleus muscles of the diabetic rats metformin treatment caused no further depolarisation. The muscles of metformin-treated control and diabetic rats hyperpolarised in the presence of insulin. Diabetes caused an increase in the volume of the blood erythrocytes. This was prevented by metformin treatment.

After 2 months of streptozotocin-induced diabetes in rats, the membrane potential of the diaphragm muscle when measured in vitro at 30¡C was unchanged but the tetanic tension, the half-relaxation time of the isometric twitch and the fatigue resistance were each reduced. Treatment of the diabetic rats with the antihyperglycaemic agent metformin prevented the decrease in half-relaxation time and the greater degree of fatigue in the diaphragms. The possibility that changes in H+ and cyclic AMP concentrations in the diabetic muscles contributed to the decreased contractile function and that metformin acted by attenuating these changes is discussed.

A comprehensive understanding of the mechanisms by which agonists control uterine contraction is essential for the successful clinical management of parturition and for the timely treatment of situations involving inappropriate uterine performance. In this review we discuss some of the key stimulatory mechanisms linking receptor occupation at the myometrial plasma membrane with alteration of myofilament activation. We focus on evidence that receptor-induced membranous recruitment of the small G-protein rhoA, and its downstream effector rho-associated kinase (ROK) is crucial to agonist-induced Ca2+-sensitisation of uterine contraction and that co-ordination of this signal transduction pathway may be mediated by the actions of caveolins, proteins integral to specialised membranous regions termed caveolae. Experimental Physiology (2001) 86.2, 283-288.

Worldwide, the highest morbidity and mortality results from such cardiovascular diseases as hypertension, myocardial infarction, cardiac and renal failure, as well as stroke. Since the cardiovascular system and its regulation is quite complex, study of these disorders has been grossly limited to whole organism models. As a result, in recent years, transgenic technology has played a significant role in the discovery of specific gene products for cardiovascular regulation and disease aetiology. Genetic manipulation in rats and mice has altered the expression of numerous genes. In this review, some of the important new genetically modified animals (i.e. transgenic models) with alterations in hormone and second messenger systems involved in cardiovascular regulation are summarized.

The present study was designed to investigate the effect of progesterone on the coronary circulation and to determine the mechanisms involved. In pigs anaesthetized with sodium pentobarbitone, changes in left circumflex or anterior descending coronary blood flow caused by intravenous infusion of progesterone at constant heart rate and arterial blood pressure were assessed using an electromagnetic flowmeter. In 14 pigs, infusion of 1 mg h-1 of progesterone caused an increase in coronary blood flow without affecting left ventricular dP/dtmax (rate of change of left ventricular systolic pressure) and filling pressures of the heart. In a further four pigs, this vasodilatory coronary effect was enhanced by graded increases in the dose of the hormone of between 1, 2 and 3 mg h-1. The mechanisms of the above response were studied in the 14 pigs by repeating the experiment after haemodynamic variables had returned to the control values observed before infusion. In six pigs, blockade of muscarinic cholinoceptors and adrenoceptors with atropine, propranolol and phentolamine did not affect the coronary vasodilatation caused by progesterone. In the remaining eight pigs, this response was abolished by intracoronary injection of Nω-nitro-L-arginine methyl ester (L-NAME) even when performed after reversing the increase in arterial blood pressure and coronary vascular resistance caused by L-NAME with continuous intravenous infusion of papaverine. The present study showed that intravenous infusion of progesterone primarily caused coronary vasodilatation. The mechanism of this response was shown to involve the endothelial release of nitric oxide. Experimental Physiology (2001) 86.1, 101-108.

It has previously been established that a single systemic administration of retroviral vector containing angiotensin II type I receptor antisense (AT1R-AS) in the neonatal spontaneously hypertensive rat (SHR) prevents development of hypertension, and in addition cardiac hypertrophy and endothelial dysfunction. However, these studies could not determine whether the effects of AT1R-AS on high blood pressure (BP) and endothelial function were independent. Angiotensin receptor blockers have been shown to reduce BP in the L-NAME (N ω-nitro-L-arginine methyl ester hydrochloride)-induced rat model of hypertension. Our objective in the present study was to use the L-NAME model of hypertension to determine whether AT1R-AS treatment would lower high BP and attenuate cardiac hypertrophy under conditions of permanent endothelial damage. A single bolus of LNSV-AT1R-AS viral particles in neonatal Wistar-Kyoto (WKY) rats was without affect on basal BP. Efficacy of the transgene incorporation was assessed by observing a significant reduction in angiotensin-induced dipsogenic response in the AT1R-AS-treated animals. Introduction of L-NAME in the drinking water for 10 weeks resulted in the establishment of hypertension only in the WKY rats treated with vector alone. These hypertensive (BP, 179 ± 4 mmHg) animals showed a 17 % increase in heart weight/body weight ratio and a 60 % reduction in ACh-induced vasorelaxation in phenylephrine-preconstricted arteries. The L-NAME-induced high BP and cardiac hypertrophy were attenuated in rats expressing AT1R-AS. However, endothelial dysfunction could not be prevented with the antisense therapy. These observations demonstrate that attenuation of endothelial dysfunction is not a prerequisite for the antihypertensive effects of AT1R-AS treatment. Experimental Physiology (2003) 88.4, 467-473.

Carbohydrate ingestion increases both endurance time to exhaustion during prolonged exercise, and the ability to perform resistance exercise. The mechanism(s)underlying the increased performance following glucose ingestion remain(s) unclear. The purpose of the present study was to verify the hypothesis that glucose infusion could attenuate peripheral muscle fatigue in the anaesthetized rat during prolonged indirect electrical stimulation in situ. For this purpose the plantaris muscle was electrically stimulated (50 Hz for 200 ms every 2.7 s; 5 V; pulse width, 0.05 ms) in situ through the sciatic nerve to perform concentric contractions for 60 min while infusing intravenously either saline alone (7.25 ml kg-1 h-1), or saline and glucose (1 g kg-1 h-1: plasma glucose 11 ± 1.1, vs. 4.9 ± 0.2 mM with infusion of saline) (8 rats per group). Glucose infusion attenuated the reduction in submaximal peak dynamic force (55 % decrease vs. 70 % decrease in rats infused with saline alone, P < 0.05). In a third group of rats (n = 8), infusion of glucose 30 min after the start of stimulation partially restored submaximal peak dynamic force (P < 0.05). Maximum dynamic and isometric forces at the end of the period of stimulation were also higher (P < 0.05) in rats infused with glucose (4.0 ± 0.2 and 4.3 ± 0.2 N, respectively) than saline alone (3.0 ± 0.2 and 3.5 ± 0.2 N, respectively). The beneficial effect of glucose infusion on peripheral muscle force during prolonged stimulation was not associated with a reduction in muscle glycogen utilisation, nor with a reduction of fatigue at the neuromuscular junction, as assessed through maximal direct muscle stimulation (200 Hz for 200 ms; 150 V; pulse width, 0.05 ms). However, changes in M-wave peak-to-peak amplitude, duration and total area suggest that glucose infusion, and/or the associated increase in plasma insulin concentration, may prevent the deterioration of electrical properties of the muscle fibre membrane. Experimental Physiology (2002) 87.5, 585-592.

The relationship between muscle force and ankle position during isometric and pre-loaded slow concentric contractions (angular velocity, 0.52 rad s-1; range of motion, 1.22 rad) and the recovery of isometric force following concentric contractions at different velocities were determined for electrically stimulated plantar flexor muscles in intact rats. Pre-loaded refers to the isometric contraction which immediately precedes the concentric contraction. Ankle position was controlled by a dynamometer and force was recorded under the sole of the foot. The peak isometric force (19.2 N) was nearly constant at all ankle positions (range of motion, 1.57 rad). The muscle length and distal fibre length of gastrocnemius medialis at ankle positions between 0.79 rad and 2.01 rad were increased by 12.6 % and 20.3 %, respectively. During slow concentric contractions, the force progressively decreased (23.1 ± 2.1 %); the force decreased by only 6.3 ± 0.9 % during sustained isometric contractions of similar duration (3400 ms). The recovery of isometric force following concentric contractions with similar stimulation frequencies (80 Hz) was velocity dependent (i.e. more rapid at higher velocities). It is concluded that pre-loaded slow concentric contractions of the plantar flexor muscles in intact rats do not follow the same relationship as that of isometric force and ankle position. Our results in intact rats show that the force output of electrically stimulated ankle plantar flexor muscles measured under the sole of the foot can be used to study the physiological properties of skeletal muscle working in situ.

The effects of Ba2+, Mg2+, Ca2+ and Na+ as blocking ions were investigated in 90 and 10 mM extracellular K+ solutions on the cloned inward rectifying K+ channel Kir2.1 expressed in Xenopus oocytes. Some data were also obtained using another inward rectifying K+ channel Kir3.1/Kir3.4. The addition of Ba2+ caused a concentration-, voltage- and time-dependent block of both channels. Decreasing the extracellular K+ concentration augmented the block. The data suggest that Ba2+ blocks the channels by binding to a site within the channel pore and that the electrical binding distance, δ, of the site is significantly different for Kir2.1 and Kir3.1/Kir3.4 ([eqv] 0·38 and [eqv] 0·22, respectively). Mg2+ and Ca2+ caused an instantaneous concentration- and voltage-dependent block of both channels. With Kir2.1, decreasing the K+ concentration augmented the block. The voltage dependence of the block was less than that of Ba2+ (δ, [eqv] 0·1), indicating a more superficial binding site for these ions within the channel pore. The affinity of the channels for Mg2+ and Ca2+ was [eqv] 1000-fold lower than that for Ba2+. Addition of Na+ resulted in a concentration-, voltage- and time-dependent block of Kir2.1, similar to that observed with Ba2+. The competition between the blocking cations (for Kir2.1: Ba2+, Mg2+, Ca2+; for Kir3.1/Kir3.4: Ba2+) and extracellular K+ suggests that the binding sites for the blocking cations may be sites to which K+ binds as part of the normal passage of K+ through the channels. It is possible that under normal physiological conditions naturally occurring extracellular cations may partly block the two inward rectifying K+ channels.

Cross-sectional areas and succinate dehydrogenase (SDH) activities of type identified fibres in the deep, middle and superficial regions of the tibialis anterior muscle in mice were examined after 4 weeks of voluntary running exercise with increasing loads. Nineteen-week-old male mice were assigned randomly to either a control or exercise group. The mean cross-sectional areas of all types (IIa, IIx and IIb) of fibres in the superficial region of the muscle were greater in the exercise group than in the control group. The mean SDH activities of type IIx and type IIb fibres in the middle region and of all types (IIa, IIx and IIb) of fibres in the superficial region of the muscle were greater in the exercise group than in the control group. These results suggest that voluntary running exercise with increasing loads causes hypertrophy and/or an increase in the SDH activity of fibres in the specific muscle region where fibres with a high threshold and a low-oxidative enzyme activity are distributed, and these fibres are recruited to adapt to changes in exercise conditions. Experimental Physiology (2002) 87.2, 113-116.

Confocal imaging in living brain slices allows the resolution of submicrometre structures of nerve cells, glia and brain vessels. Imaging living brain slices is in many respects different from conventional fixed histological preparations for which confocal microscopes were designed originally. Several problems (i.e. mechanical and thermal drift, and autofluorescence) resulting from the optical and structural properties of brain slices are discussed. Fluorescent indicators may be used to monitor numerous intracellular parameters such as pH and Ca2+ concentration, but not all of them are equally suitable for this type of work. Genetically engineered fluorescent proteins can be used to visualise the fine dendritic structure of neurones or track particular intracellular structures and proteins. They have also been used to generate indicators for Ca2+, cAMP and other molecules. While conventional chemical indicators can be either loaded into neurones via patch pipettes or as membrane-permeable esters, protein indicators can be expressed in various types of cells using adenoviral vectors. Adenoviral transgenesis can be performed in vitro in both acute slices and organotypic slice cultures. Organotypic slice cultures give excellent optical access to neurones loaded with either conventional fluorescent indicators or transfected with adenovirus to express fluorescent proteins. They are most suitable for experiments where both conventional and genetically engineered indicators are combined. Single photon imaging in brain slices is limited to the superficial layers (∼≤50 mm), while multiphoton excitation has a much greater depth of penetration. However, the overall optical resolution achievable in single photon mode is at least as good as when using multiphoton excitation.

Maternal and fetal body temperatures were measured in five Boer goats, of mean mass 64 ± 8 kg, using temperature-sensitive radiotelemeters implanted intra-abdominally. Body temperatures were recorded every 5 min. Throughout the last month of gestation, fetal temperature was approximately 0.6 oC higher than that of the mother, in normal laboratory conditions (ambient air temperature: 21-24 oC). This feto-maternal temperature difference between the goat fetus and its mother is similar to that found in other mammals, including sheep. When the pregnant goats were subjected to short-term heating and cooling, the difference between maternal and fetal body temperatures changed. Thus the mean difference between fetal and maternal body temperatures decreased from 0.4 to 0.2 oC during 2 h of heating, while it widened from 0.3 to 0.7 oC during 6 h of cooling. These data support the idea that the fetus is thermally protected from excursions of body temperature during changes in the mother's thermal environment. Reports of goat stock losses and abortions during cold spells in their natural habitats may be the result of more severe and/or prolonged cold exposures that not only adversely affect fetal or maternal body temperature, but also influence other aspects of metabolism. Experimental Physiology (2001) 86.2, 199-204.

It is known that arterial baroreflexes are suppressed in stressful conditions. The present study was designed to determine whether and how hypoxia affects arterial baroreflexes, especially the heart rate component, baroreflex vagal bradycardia. In chloralose-urethane-anaesthetized rats, baroreflex vagal bradycardia was evoked by electrical stimulation of the aortic depressor nerve, and the effect of 15 s inhalation of hypoxic gas (4 % O2) was studied. Inhalation of hypoxic gas was found to inhibit baroreflex vagal bradycardia. The inhibition persisted after bilateral transection of the carotid sinus nerve. Cervical vagus nerves were cut bilaterally and their peripheral cut ends were stimulated to provoke vagal bradycardia of peripheral origin so as to determine whether hypoxia could inhibit vagal bradycardia by acting on a peripheral site. In contrast to baroreflex vagal bradycardia, the vagus-induced bradycardia was not affected by hypoxic gas inhalation. It is concluded that baroreflex vagal bradycardia is inhibited by hypoxia and the inhibition is largely mediated by its direct central action.

Feeding a high-carbohydrate (CHO) diet and administration of alkalinizing agents have both been shown to improve performance in high-intensity exercise. The effect of these treatments in combination was investigated in the present study. Six healthy male subjects exercised to exhaustion on an electrically braked cycle ergometer at a power output equivalent to 100 % of their maximum oxygen uptake (VO2,max) on four separate occasions. Each subject consumed either a diet with the same composition as his normal diet (termed the experimental normal (N) diet; 54 ± 7 % CHO, 13 ± 2 % protein, 33 ± 7 % fat) or a high-CHO diet (81 ± 2 % CHO, 13 ± 2 % protein, 6 ± 1 % fat) that had the same energy and protein content for the 3 days prior to the exercise tests. Subjects then ingested either a placebo (CaCO3) or trisodium citrate (0·3 g (kg body mass)-1) 3 h before exercise. Time to fatigue was not different between experimental conditions. Consumption of the high-CHO diet had no effect on blood acid-base status, but the ingestion of sodium citrate induced a mild metabolic alkalosis after both the N diet and the high-CHO diet. This alkalinizing effect was also evident after exercise, since blood pH, plasma bicarbonate and blood base excess were higher (P < 0·05) after the ingestion of sodium citrate than under the placebo conditions. The changes in blood lactate, pyruvate and glucose and plasma glycerol after exercise were similar for all experimental conditions. Blood lactate, glucose and pyruvate and plasma glycerol concentrations increased from resting values (P < 0·01) following exercise but this increase was similar under all experimental conditions. These data demonstrate that when the energy and protein content of the diets is the same, exercise capacity and the metabolic response to intense exercise are similar following consumption either of a high-CHO diet or a more normal diet. Acute ingestion of sodium citrate prior to exercise resulted in a reduction in post-exercise acidosis despite a blood lactate concentration that was similar to that observed after the ingestion of a placebo, but did not affect exercise performance under the conditions of this study.