Muscle vasodilatation evoked by systemic hypoxia is adenosine mediated and nitric oxide (NO) dependent: recent evidence suggests the increased binding of NO at complex IV of endothelial mitochondria when O2 level falls leads to adenosine release. In this study on anaesthetised rats, the increase in femoral vascular conductance (FVC) evoked by systemic hypoxia (breathing 8 % O2 for 5 min) was reduced by oxypurinol which inhibits xanthine oxidase (XO): XO generates O2- from hypoxanthine, a metabolite of adenosine. By contrast, infusion of superoxide dismutase (SOD), which dismutes O2- to hydrogen peroxide (H2O2), potentiated the hypoxia-evoked increase in FVC. However, NO synthesis inhibition reduced the hypoxia-evoked increase in FVC and it was not further altered by SOD. In other studies, the spinotrapezius muscle was pre-loaded with hydroethidine (HE), or dihydrorhodamine (DHR) which fluoresce in the presence of O2- and H2O2, respectively. In muscle loaded with HE, systemic hypoxia increased fluorescence in endothelial cells of arterioles, whereas in muscle loaded with DHR, fluorescence was diffusely located in and around arteriolar endothelium. We propose that in systemic hypoxia, O2- generated by the XO degradation pathway from adenosine released by endothelial cells, and released by endothelial mitochondria by increased binding of NO to complex IV, is dismuted to H2O2, which facilitates hypoxia-induced dilatation. Experimental Physiology (2003) 88.6, 733-740.

Mammals that use the ventilatory system as the principal means of increasing heat loss, i.e. that pant, show two fundamental changes in the control of breathing at raised temperatures. First, alveolar ventilation increases by more than, rather than in proportion to, the increase in CO2 production. Second, hypocapnia no longer causes apnoea. Rats do not use the ventilatory system as the principal means of increasing heat loss, so we have investigated whether rats also show these two changes at raised temperatures. Breathing was detected from diaphragmatic electromyogram (EMG) activity. Anaesthesia and hyperoxia were used to minimise behavioural and hypoxic drives to ventilation and arterial PCO2 (Pa,CO2) was controlled using mechanical ventilation. At 36.6 ± 0.1 °C, breathing was absent as long as Pa,CO2 was held below a threshold level of 32.9 ± 0.7 mm Hg (n = 14) under steady-state conditions. When body temperature in rats was raised above 37 °C, both fundamental changes in the control of breathing became apparent. First, at 39 °C the mean Pa,CO2 level during spontaneous breathing (39.6 ± 5.4 mm Hg, n = 4) fell by 3.9 ± 1.4 mm Hg (P < 0.05, Student's paired t test). Second, at 39.9 ± 0.1 °C breathing was present when mean Pa,CO2 levels were only 18.2 ± 1.5 mm Hg (n = 14), the lowest mean Pa,CO2 level we could achieve with mechanical ventilation. We calculate, however, that at 39.9 °C, the drive to breathe from the increased CO2 production alone would not sustain breathing below a Pa,CO2 level of 27.8 ± 1.4 mm Hg (n = 13). In rats at raised body temperatures therefore a respiratory drive exists that is in addition to that related to the increase in CO2 production. Experimental Physiology (2000) 85.3, 309-319.

Chronic stimulation of β2-receptors with β2-agonists causes desensitisation, which in skeletal muscle is accompanied by myosin heavy chain (MHC) remodelling, similar to that observed in heart failure patients. However, the mechanisms for this skeletal muscle remodelling are not well established. G protein-coupled receptor kinases (GRKs) specifically phosphorylate and desensitise G protein-coupled receptors during periods of agonist activation. However, desensitisation associated with prolonged agonist activation alters β-adrenergic signalling, and downstream affects gene expression. We hypothesised that skeletal muscle remodelling induced by β2-agonist administration could be regulated by GRK expression. Therefore the aim of this study was firstly to characterise which, if any, of the six known isoforms of GRK were expressed in skeletal muscle and then secondly to determine whether remodelled skeletal muscle induced by chronic β2-agonist administration was accompanied by altered expression of GRK isoforms. Male Wistar rats were administered a β2-agonist daily for 8 weeks, and the expression of MHC and GRKs examined in gastrocnemius and soleus muscles. Treatment with β2-agonist caused a change in MHC in soleus from types I to IIA, and in gastrocnemius from MHC types IIA/IIX to IIB. Western blotting revealed that GRK2 and GRK5 were expressed in skeletal muscle. Furthermore, despite changes in MHC and differential muscle-specific expression of GRK isoforms, there was no significant change in expression of GRK2 and GRK5 in soleus or gastrocnemius following β2-agonist administration. In conclusion the level of GRK expression is unlikely to be responsible for MHC switching following chronic β2-receptor stimulation. Experimental Physiology (2003) 88.2, 277-284.

The renal response to arginine vasopressin in the rat has been shown to depend on reproductive status. However there is no consensus as to when the kidney is most responsive. The varying results could depend on the protocol and the dose of hormone used. A study has been performed, with physiological doses of vasopressin, comparing the responses during infusion of hypotonic saline and glucose. After an equilibration period of 150 min, conscious rats were infused on each of the four days of the oestrous cycle with either isotonic saline (0.077 M) or 0.14 M glucose for a control period of 45 min. Vasopressin was then infused at 10-40 fmol min-1 for 1 h, followed by a recovery period of 90 min. Timed urine samples were collected for determination of volume, sodium concentration and osmolality. During the control period urine flow was greatest at oestrus and dioestrus day 2 and sodium excretion on dioestrus day 2 irrespective of the infusate. Vasopressin concentrations achieved lay within the physiological range and no difference was observed between the different days for a given dose. Infusion of vasopressin in both saline and glucose produced a dose-dependent antidiuresis, the greatest responses being seen of pro-oestrus and dioestrus day 2. It was only with the highest rate of infusion that a significant increase in sodium excretion was seen on each day of the cycle and the greatest responses were seen on pro-oestrus and dioestrus day 1 for both infusates. Thus the kidney shows the greatest response to physiological doses of vasopressin at pro-oestrus and dioestrus day 1 irrespective of the infusate employed. Experimental Physiology (2002) 87.1, 9-15.

Intravenous injection of growth hormone in anaesthetized pigs has been shown to cause coronary vasoconstriction by antagonizing the vasodilatory effects of β2-adrenergic receptors. Because nitric oxide is believed to modulate or mediate β2-adrenergic effects, the present study was undertaken in the same experimental model to determine the role of nitric oxide in the above response to growth hormone. In fourteen pigs anaesthetized with sodium pentobarbitone, changes in left circumflex or anterior descending coronary blood flow caused by intravenous injection of 0.05 i.u. kg-1 of growth hormone at constant heart rate and arterial blood pressure were assessed using electromagnetic flowmeters. In a first control group of six pigs, growth hormone caused a decrease in coronary blood flow which averaged 13.1 % of the baseline values. In a second group of eight pigs, intravenous administration of Nω-nitro-L-arginine methyl ester (L-NAME) was used to block the endothelial release of nitric oxide. In these pigs, the subsequent injection of growth hormone did not cause any significant changes in coronary blood flow, 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. These results indicated that the coronary vasoconstricting effect of growth hormone, known to involve antagonism of β2-adrenergic vasodilatory effect, was mediated by inhibition of nitric oxide release.

In the past several years, the progress in the development of molecular biology and the elucidation of gene sequences has created new approaches for studying biological functions and developing new diagnostic and therapeutic strategies. One of the most exciting advances has been the development of antisense technology, which represents a new strategy allowing modulation of protein synthesis with high specificity by preventing protein expression at the level of RNA or DNA. Many classical pharmacological approaches in neurobiological research are often based on the inhibition of biologically active proteins, such as receptors for neurotransmitters, or enzymes involved in neurotransmitter synthesis or degradation. The use of antisense oligodeoxynucleotides offers an alternative tool to manipulate selectively the expression of neurotransmitters or their receptors in neuronal tissue. This approach is especially useful when selective, high-affinity antagonists are not available. As a result, this technology has gained acceptance in the study of cell signalling mechanisms and the molecular basis of neuronal function. This paper provides a brief background to the antisense technique and explores methodological aspects, particularly in the whole animal. The use of the antisense technology in studies focused on central mechanisms regulating the cardiovascular system is then discussed.

The purpose of this study was to determine the influence of testosterone, the male sex hormone, on β-adrenergic agonist-induced amylase secretion from rat parotid glands. Isoprenaline (isoproterenol)-induced amylase secretion was measured in vitro from the parotid glands of control and castrated rats with and without testosterone replacement. The isoprenaline-induced amylase release was reduced in parotid glands from castrated rats compared to controls. The reduction of amylase release by isoprenaline in parotid glands of castrated rats, could be reversed by administration of testosterone. Furthermore, β-adrenergic receptor density and the level of isoprenaline-evoked cAMP in parotid glands from castrated rats was lower compared to intact rats. Using SQ-22536 (an adenylyl cyclase inhibitor), dibutyryl cAMP (a cAMP analogue) and verapamil (a calcium channel blocker), we conclude that the impairment of amylase release from parotid glands after castration was not related to either adenylyl cyclase activity or cAMP accumulation. Amylase release from the parotid glands of castrated rats appears to be mediated by an increase in calcium ion influx.

The extrusion mechanism for intracellular Mg2+ was investigated in voltage-clamped snail neurones using Mg2+-sensitive microelectrodes and mag-fura-2. The intracellular free magnesium ion concentration ([Mg2+]i) of snail neurones voltage clamped to -60 mV was estimated to be 0·57 ± 0·06 mM (mean ± S.E.M.; n = 12) using Mg2+-sensitive microelectrodes and 0·62 ± 0·05 mM (n = 15) using mag-fura-2 . Raising extracellular MgCl2 from 5 to 20 mM caused an average increase in [Mg2+]i of 0·25 ± 0·04 mM (n = 7). In three experiments, removing extracellular MgCl2 caused an average decrease in [Mg2+]i of 0·1 mM. Replacing extracellular Na+ with N-methyl-D-glucamine (NMDG) caused a rise in [Mg2+]i of 1·8 ± 0·5 mM (n = 7); [Mg2+]i recovered to resting levels when extracellular Na+ was restored. Iontophoretic injections of MgCl2 were used to raise [Mg2+]i. The rate of recovery from such increases in [Mg2+]i (calculated from the slope of the recovery) was inhibited by 85-100 % (n = 5) in the absence of extracellular Na2+ compared with control conditions. Raising extracellular Ca2+ from 7 to 35 mM caused a reversible rise in [Mg2+]i of 0·4 ± 0·05 mM (mean ± S.E.M., n = 7). It was concluded that in snail neurones the main mechanism for [Mg2+]i extrusion is a Na+-Mg2+ exchanger which may be partially inhibited be high extracellular Ca2+ concentrations.

The effect of stimulation frequency on intact muscle blood flow was measured by radiolabelled microsphere entrapment. Maximum flow occurred at 4 Hz in both extensor digitorum longus and tibialis anterior (156 ± 13 and 205 ± 24 ml min-1 (100 g)-1, respectively). Regional differences in fibre composition were reflected in the relative flow rates, with functional hyperaemia in the oxidative tibialis anterior core being twice that of the glycolytic cortex at 10 Hz. A consistent difference in flow between legs was noted that may originate from an unequal division of the common iliac artery in rats.

Experiments were performed in the normal rat knee joint to investigate the role of different isoforms of cyclooxygenase (COX) in the regulation of basal joint blood flow. Laser Doppler imaging (LDI) was used to measure articular perfusion, and reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of COX-1 and COX-2 mRNA in joint tissue. Intravenous infusion of indomethacin (a non-selective inhibitor of COX; 0.34 nmol min-1) over 40 min produced a time dependent increase in articular vascular resistance (maximum 22.5 % at 40 min; P < 0.0001, one-way ANOVA) whereas vehicle over a similar time period had no effect in a control group. An equimolar concentration of a highly selective inhibitor for COX-2, SC-236, was administered in a further group of rats but this did not increase articular vascular resistance. While there was no significant difference between the response to vehicle and SC-236 (two-way ANOVA; P = 0.686, n = 6) the response to indomethacin was significantly greater than vehicle or SC-236 (two-way Anova; P < 0.0001, n = 6). COX-1, but not COX-2, was detectable by RT-PCR in all joint tissue samples examined (n = 4). The results of this study indicate that prostaglandins (PGs) play an important role in the maintenance of basal perfusion in the rat knee joint, with COX-1 being the physiologically relevant isoform. Experimental Physiology (2001) 86.2, 191-197.

An investigation was undertaken into the potential role of nitric oxide (NO) and its interaction with renal sympathetic nerves in mediating renal responses to acute saline volume expansion (VE). Groups of anaesthetized Wistar rats with innervated and denervated kidneys were subjected to VE, 0·25 % body wt min-1 for 40 min, in the presence and absence of nitric oxide synthase (NOS) inhibitors, NG-nitro-{fontsize L-arginine-methyl-ester ({fontsize L-NAME, non-selective), aminoguanidine (AG, relatively selective for inducible NOS (iNOS)), and 7-nitroindazole (7-NI, relatively selective for neuronal NOS (nNOS)). Pretreatment with {fontsize L-NAME or AG enhanced the cumulative sodium excretion (CuUNaV) after 40 min VE in the innervated kidneys by 27 and 23 % (both P < 0·001), respectively, compared to the untreated control group, whereas they were without effect in the denervated kidneys. Cumulative urine flow (CuUV) after VE in {fontsize L-NAME- and AG-treated groups was enhanced in both kidneys, by some 17-21 % in the denervated (P < 0·01) and 37-39 % in the innervated kidneys (P < 0·001) by comparison with the corresponding untreated controls. 7-NI had no effect on CuUV, but reduced CuUNaV in the denervated kidneys by 25 % (P < 0·01) when compared to the control group. The results suggested that NO, possibly generated by endothelial NOS (eNOS) and iNOS, was a contributory factor in mediating the renal response to VE. There appeared to be a tonic inhibitory action of NO on water excretion which was renal nerve independent, whereas its impact on sodium handling appeared to be dependent upon a background level of renal nerve activity. Experimental Physiology (2001) 86.1, 47-54.

In order to describe the factors which, during hyperoxic exercise, can counteract the chemoreceptor-mediated inhibition of ventilation by O2, minute ventilation (VE) and the pulmonary gas exchange were studied breath-by-breath in four dogs running on a treadmill (5 km h-1) for 10 min during and following exposure to O2 of different durations. We found that a brief inhalation of O2 applied during the steady state of the VE response provoked a reduction in VE by 6.5 ± 0.9 l min-1 whereas hyperoxia applied 2 min before the onset of exercise and maintained for 2.5 min during the running tests had a significantly weaker effect on VE (-1.8 ± 0.2 l min-1, P < 0.05). The rise in pulmonary CO2 output (VCO2) during the prolonged O2 exposure was less than in normoxic exercise leading to a deficit of CO2 eliminated by the lungs of 181 ml. The return to air breathing provoked a rise in VE, which reached within 73 s a much higher level than the control tests (22.9 ± 3.6 vs. 19.5 ± 2.2 l min-1, P < 0.05); VE then subsided to control levels with a long exponential decline. The CO2 deficit during O2 breathing, was fully compensated after recovery in air within 6 min. No stimulatory effect on ventilation was observed at rest at the cessation of a similar exposure to O2 despite a higher end-tidal PCO2 (+4 ± 1 mmHg) than in exercise. In conclusion, the stimulatory effect of O2 during exercise can be clearly revealed after recovery in air and seems to operate through a more complex mechanism than that thought to be involved at rest. We propose that the changes in CO2 stores in the exercising muscles could contribute to O2-induced stimulation during exercise, possibly through stimulation of muscle afferents responding to local circulatory changes. Finally, the observation that during continuous dopamine (DA) infusion (5 µg kg-1 min-1) the VE response to recovery in air was only a slow decrease, suggests that the arterial chemoreceptors potentiate O2-induced hyperventilation, or that the vascular actions of DA counteract part of the effects provoked by CO2 accumulation in the exercising muscles.

Four adult Corriedale sheep were used in an experiment divided into three parts. In part 1 a primed continuous infusion of [6,6-2H2]glucose was infused for 7 h. The first 3 h was the control period, from 3 to 7 h glucose-dependent insulinotropic polypeptide (GIP) was infused, and from 5 to 7 h somatostatin was infused. Part 2 of the experiment was the same as for part 1 except that insulin was infused between 3 h and 7 h and GIP was infused between 5 and 7 h. Coincident with the insulin infusion, normal glucose was also infused at a variable rate in order to keep the plasma glucose at basal levels. In part 3 of the experiment [6,6-2H2]glucose was infused for 5 h and somatostatin was infused between 3 and 5 h. Measurements of glucose turnover were made in the last 40 min of the control, GIP only, insulin only, somatostatin only, GIP plus somatostatin and GIP plus insulin infusion periods. Plasma insulin levels were reduced to the limit of detection by the somatostatin infusion; under such conditions whole-body glucose uptake should be entirely non-insulin-mediated (NIMGU). Expressing glucose disposal as glucose metabolic clearance rate demonstrated that elevated, but still physiological GIP levels had no effect on NIMGU but significantly increased insulin-mediated glucose uptake when plasma insulin levels were similar to levels typically observed after a meal. These results indicate that in sheep, GIP may enhance insulin action with respect to glucose disposal following a meal, but has no effect on glucose disposal pathways not responsive to insulin.

This study was undertaken to improve the measurement of glomerular filtration rate (GFR) during the acute diuretic phase induced by atrial natriuretic peptide (ANP), which may indeed alter the renal clearance of inulin (GFRCL) due to dead space error. A technique to measure GFR without urine collections was therefore developed in anaesthetized rats prepared as for micropuncture. To do so, arterial blood was periodically collected and renal venous blood was withdrawn simultaneously from a catheter inserted into the left suprarenal vein to determine the renal extraction coefficient of inulin (CEIN). In addition, renal blood flow (RBF) was continuously measured with an electromagnetic flow transducer fitted around the left renal artery to estimate renal plasma flow (RPF). GFR (GFRCE) was then calculated as the product of RPF and CEIN. To study the effects of ANP on GFR, rats were injected I.V. with 10 µl of saline without (n = 6; vehicle) or with 1 µg ANP (n = 6; ANP) and GFRCE and GFRCL were compared before and after each treatment. They did not differ significantly during baseline measurements in each experimental group and were not modified after vehicle. Similarly, RBF remained constant. In contrast, RBF and GFRCE increased rapidly and simultaneously 90 s after ANP, from 9á07 ± 0á25 to 10á07 ± 0á35 (12 %) and from 1á209 ± 0á188 to 1á715 ± 0á190 ml min-1 (42 %), respectively (P < 0á05). GFRCL increased to an even greater extent (88 %). Moreover, the peak enhancement of GFRCL was delayed and occurred 180 s after ANP. The renal clearance of inulin was thus unduly elevated due to sudden changes in the dead space induced by the diuretic effect of ANP. In conclusion, determination of glomerular filtration rate by the method of renal extraction of inulin provided more reliable results than those achieved using the classical method of renal clearance of inulin. Moreover, it was sufficiently sensitive to detect small and transient changes in GFR induced by the injection of 1 µg ANP.

We investigated anaerobic threshold (thetaL) gas exchange kinetics and maximal oxygen uptake (VO2,max) among older men with reduced left ventricular end-diastolic filling (LVDF). Ten men (mean age, 73 years) with LVDF impairment and low fitness, but without other cardiovascular dysfunction were studied. Treatments compared to control included: 5 days, high intensity exercise training protocol; 5 days, calcium channel blockade (240 mg verapamil); 21 days, detraining/washout; and 5 days, combined treatments. Results indicated no changes in resting left ventricular systolic function with any treatment. Significant resting diastolic function changes included increased early:late flow velocity (control, 0.87; training, 1.28; verapamil, 1.32), and a decreased isovolumic relaxation time (control, 0.10 s; training, 0.08 s; verapamil, 0.08 s). The combined treatments were not additive. Sub-threshold oxygen uptake kinetics (tauVO2, s) were significantly faster following either training or verapamil (tauVO2,control, 62 ± 12; tauVO2,training, 44 ± 9; tauVO2,verapamil, 48 ± 10) and combined treatments (tauVO2, 41 ± 8). V O2,max (ml kg-1 min-1) was significantly increased (control, 21.8 ± 2.2; training, 27.3 ± 2.2; verapamil, 25.2 ± 3.4; combined treatments, 26.9 ± 2.3). Increasing ventricular preload with either exercise training or calcium channel blockade was coincident with faster tauVO2 and increased VO2,max. Experimental Physiology (2000) 85.5, 547-555.

The dynamics and kinetics of thyroid hormone transport in the isolated rat heart were examined using the modified unidirectional paired tracer dilution method. The uptake of 125I-thyroxine (125I-T4) and 125I-triiodothyronine (125I-T3) from the extracellular space into heart cells was measured relative to the extracellular space marker 3H-mannitol. The thyroid hormone maximal uptake was 54.4 % for 125I-T4 and 52.15 % for 125I-T3. The thyroid hormone net uptake was 25.69 % for 125I-T4 and 25.49 % for 125I-T3. Backflux from the intracellular space was 53.17 % for 125I-T4 and 61.59 % for 125I-T3. In the presence of unlabelled thyroid hormones, 125I-T4 and 125I-T3 maximal uptakes were reduced from 10.1 to 59.74 % and from 34.6 to 65.3 %, respectively, depending on the concentration of the unlabelled hormone, suggesting a saturable mechanism of the thyroid hormone uptake by the heart cells, with Km(T4)= 105.46 µM and the maximal rate of 125I-thyroid hormone flux from the extracellular space to heart cells (Vmax(T4)) = 177.84 nM min-1 for 125I-T4 uptake, and Km(T3) = 80.0 µM and Vmax(T3) = 118.5 nM min-1 for 125I-T3 uptake. Experimental Physiology (2001) 86.1, 13-18.

This study examined the effect of pre-exercise carbohydrate (CHO) ingestion on pyruvate dehydrogenase complex (PDC) activation, acetyl group availability and substrate level phosphorylation (glycogenolysis and phosphocreatine (PCr) hydrolysis) in human skeletal muscle during the transition from rest to steady-state exercise. Seven male subjects performed two 10 min treadmill runs at 70 % maximum oxygen uptake (VO2,max), 1 week apart. Each subject ingested 8 ml (kg body mass (BM))-1 of either a placebo solution (CON trial) or a 5.5 % CHO solution (CHO trial) 10 min before each run. Muscle biopsy samples were obtained from the vastus lateralis at rest and immediately after each trial. Muscle PDC activity was higher at the end of exercise in the CHO trial compared with the CON trial (1.78 ± 0.18 and 1.27 ± 0.16 mmol min-1 (kg wet matter (WM))-1, respectively; P 0.05) and this was accompanied by lower acetylcarnitine (7.1 ± 1.2 and 9.1 ± 1.1 mmol kg-1 (dry matter (DM))-1 in CHO and CON, respectively; P 0.05) and citrate concentrations (0.73 ± 0.05 and 0.91 ± 0.10 mmol (kg DM)-1 in CHO and CON, respectively; P 0.05). No difference was observed between trials in the rates of muscle glycogen and PCr breakdown and lactate accumulation. This is the first study to demonstrate that CHO ingestion prior to exercise augments the exercise-induced activation of muscle PDC and reduces acetylcarnitine accumulation during the transition from rest to steady-state exercise. However, those changes did not affect the contribution of substrate level phosphorylation to ATP resynthesis. Experimental Physiology (2000) 85.5, 581-586.

Control (normothermic) and cold-acclimated (environmental temperature gradually reduced from 20 to 5 °C for 4 weeks) groups of male rats and hamsters were compared to elucidate the nature of angiogenesis in oxidative and glycolytic muscles of these species during progressive cold exposure. Skeletal muscle capillarity and fibre cross-sectional area were measured in the tonic soleus (SOL) and phasic tibialis anterior (TA). Cryostat sections were stained for alkaline phosphatase (ALP) activity to identify all capillaries, and proliferating cell nuclear antigen (PCNA) to localise the site of cellular proliferation. Cold-induced angiogenesis, indicated by an increase in capillary to fibre ratio (C:F), occurred in SOL of rats (~20 % increase, P < 0.05) but not hamsters (~9.5 % increase, n.s.), and in TA of hamsters (~22 % increase, P < 0.01) but not rats (~1 % increase, n.s.). The change in C:F was highest in the glycolytic cortex region of TA where fibre size is larger than in the oxidative core. Capillary-specific cell proliferation (co-localised ALP and PCNA labelling) increased in parallel with C:F. The total PCNA label density within the interstitium was some 5-fold higher than that co-localised with capillaries, but where angiogenesis occurred the relative increase in capillary labelling was 2-fold greater than for other cells of the interstitium. These data suggest a significant role for endothelial cell proliferation in the angiogenic response, indicative of the sprouting form of angiogenesis. There was a tendency for fibre hypertrophy in both SOL and TA of rats, especially in the core region of TA (P < 0.01), such that capillary density (CD) and intramuscular diffusion distances (DD) were largely unchanged following cold exposure. In contrast, fibre size was maintained in hamsters, DD reduced and CD increased compared to control TA (P < 0.01). In conclusion, cold acclimation stimulated angiogenesis in muscle of hamsters more than in rats, possibly due to a higher metabolic rate in the smaller species. Angiogenesis was also seen in SOL of rat, where oxidative capacity and muscle activity is higher than the TA. Thus, a combination of oxidative capacity, muscle activity, and fibre size may determine the degree of angiogenesis in response to low environmental temperature. Experimental Physiology (2003) 88.6, 741-746.

The aim of this study was to determine the ouabain receptor density, Na+,K+-ATPase function and contractile properties of cardiac muscle in insulin-dependent and non-insulin-dependent rat diabetes mellitus (IDDM and NIDDM, respectively) and the reversibility of the diabetes-induced changes by insulin or thyroxin substitution. IDDM and NIDDM were induced in Wistar rats by streptozotocin injection. Contractile parameters were measured in isolated left ventricular trabeculae. [3H]Ouabain binding to myocardium was measured in right and left ventricular strips obtained from diabetic animals and their age-matched controls. Both the maximum [3H]ouabain binding capacity (Bmax) and the Kd for [3H]ouabain binding, as well as maximum 86Rb+ uptake and rate of contraction, were decreased in IDDM myocardium compared with controls. Insulin or thyroxin substitution reversed the reduction in Bmax and contraction rate, but not the decrease in Kd. In young, but not old, control animals, both Bmax and maximum 86Rb+ uptake were higher in the right ventricular myocardium than in the left one. In contrast to changes observed in IDDM, both Bmax and Kd for [3H]ouabain binding were increased in the left but not in the right ventricle of NIDDM animals. NIDDM caused no alterations in contractile properties. Prominent differences were observed in [3H]ouabain binding characteristics and myocardial contractility between young and old control animals. Bmax of [3H]ouabain binding and rate of contraction were inversely proportional in all preparations studied. It is concluded that IDDM and NIDDM induce different alterations in myocardial Na+,K+-ATPase, and these changes may influence the contractile properties of cardiac muscle.

Recently attempts have been made to analyse blood pressure (BP) and renal sympathetic nerve activity (RSNA) to determine how patterns contained within them might reflect control by the autonomic nervous system. To date, these studies have primarily used coherence analysis of BP and RSNA in the frequency domain. However, this analysis is unable to assess the non-linear properties of the underlying cardiovascular control system. In this study we employed not only coherence analysis but also cross-entropy analysis using balloon inflation (‘balloon-on’) to assess the influence of right atrial receptors on the relationship between BP and RSNA under two conditions in anaesthetised Wistar rats. Balloon-on stimulation alone inhibited RSNA by 28 ± 4% in eight rats without changing BP. This effect on integrated nerve activity was not present when atrial stimulation was applied during stimulation of a site in the paraventricular nucleus (PVN) of the hypothalamus which increased RSNA by 158.7 ± 58% and increased BP by 17.1 ± 2.3 mmHg. However, the cross-entropy measurement was significantly decreased (P < 0.05) during balloon-on stimulation in both the conditions revealing that there is greater synchrony between the oscillating signals contained within the BP and RSNA time series. Thus during the enhanced RSNA elicited by stimulation of the PVN, the inhibitory influence of atrial receptors, although apparently blocked, was still effective in that it resulted in the energy of the RSNA spectrum becoming more evenly distributed over a range of frequencies. The data show that cross-entropy calculations are able to characterize the non-linearities of underlying cardiovascular control.