Biomedical subjects

E Q Colquhoun

Publications and source records attributed to E Q Colquhoun.

48 records · Page 3Linked to original sources

PubMed1991

Norephedrine (phenylpropanolamine) stimulates oxygen consumption and lactate production in the perfused rat hindlimb.

d,l-Norephedrine (phenylpropanolamine) which is both a demethylated analogue and a metabolite of d,l-ephedrine, is a reputed anorectic agent. In the present study the proposed most active isomer of this mixture, l-norephedrine has been assessed as a peripherally acting thermogenic agent in the isolated perfused rat hindlimb. l-Norephedrine produced a dose-dependent increase in oxygen uptake and perfusion pressure and increased lactate production. Whereas propranolol potentiated the increase in oxygen uptake and perfusion pressure produced by l-norephedrine, prazosin significantly and nitroprusside totally inhibited both of these changes. Nitroprusside also completely inhibited the increase in lactate production. We conclude that norephedrine has a hitherto unrecognized peripheral thermogenic activity in the perfused rat hindlimb resulting from its interaction with alpha 1 adrenergic receptors that control vasoconstriction in this tissue.

Animals↗

PubMed1990

Release of purine and pyrimidine nucleosides and their catabolites from the perfused rat hindlimb in response to noradrenaline, vasopressin, angiotensin II and sciatic-nerve stimulation.

Uric acid and uracil were released at constant rates (0.95 and 0.4 nmol/min per g respectively) by the perfused rat hindlimb. Noradrenaline, vasopressin or angiotensin II further increased the release of these substances 2-5-fold, coinciding with increases in both perfusion pressure (vasoconstriction) and O2 uptake. The hindlimb also released, but in lesser amounts, uridine, hypoxanthine, xanthine, inosine and guanosine, and all but hypoxanthine and guanosine were increased during intense vasoconstriction. Uric acid and uracil releases were increased by noradrenaline in a dose-dependent manner. However, the release of these substances did not fully correspond with the dose-dependent increase in O2 uptake and perfusion pressure, where changes in the latter occurred at lower doses of noradrenaline. Sciatic-nerve stimulation (skeletal-muscle contraction) did not increase the release of uracil, uric acid or uridine, but instead increased the release of inosine (7-fold) and hypoxanthine (2-fold). Since the UTP content as well as the UTP/ATP ratio are higher in smooth muscle than in skeletal muscle, it is proposed that release of uric acid and uracil arises from increased metabolism of the respective adenosine and uridine nucleotides during intense constriction of smooth muscle.

Angiotensin II↗

PubMed1990

Inhibition by vasodilators of noradrenaline and vasoconstrictor-mediated, but not skeletal muscle contraction-induced oxygen uptake in the perfused rat hindlimb; implications for non-shivering thermogenesis in muscle tissue.

1. The effect of noradrenaline as well as of vasopressin and angiotensin II to increase oxygen uptake and perfusion pressure by the isolated perfused rat hindlimb were completely inhibited by the vasodilators, nitroprusside (0.5 mM), nifedipine (2.5 microM) and isoprenaline (50 nM). 2. Oxygen uptake due to sciatic nerve stimulation of skeletal muscle contraction was not inhibited by 0.5 mM nitroprusside but was found to increase further that produced by a maximum dose of either noradrenaline or angiotensin II. 3. Analysis of high energy phosphates in samples of freeze-clamped hindlimb muscle showed no difference before and after vasoconstrictor addition or with muscle sampled in vivo. 4. It is concluded that norepinephrine mediated increase in oxygen uptake by the perfused rat hindlimb results from its vasoconstrictor action.

Adenosine Diphosphate↗

PubMed1990

A comparison of vasopressin and noradrenaline on oxygen uptake by perfused rat hindlimb, kidney, intestine and mesenteric arcade suggests that it is in part due to contractile work by blood vessels.

1. The rat hindlimb, kidney and intestine were each perfused in a nonrecirculating mode at 25 degrees C using an artificial perfusate (initial pressure 85 +/- 5 mmHg) and the effects of vasopressin and noradrenaline on oxygen uptake and perfusion pressure determined. 2. Both vasopressin (K0.5 = 0.1 nM) and noradrenaline (K0.5 = 2 nM) increased oxygen uptake as well as perfusion pressure by the perfused hindlimb; changes in oxygen uptake were closely matched by changes in pressure. The maximum increase in oxygen uptake was approx. 9 mumol/hr per g wet wt of hindlimb. 3. The perfused kidney also responded to vasopressin and noradrenaline with parallel increases in oxygen uptake and perfusion pressure for each agent. The largest increase in oxygen uptake was approx. 30 mumol/hr per g wet wt but this was not maximal. 4. Vasopressin increased oxygen uptake and pressure by the perfused intestine over the range 0.01-2 nM, but the changes in pressure only became significant at doses greater than 0.1 nM. 5. Noradrenaline inhibited oxygen uptake and increased perfusion pressure in a dose-dependent manner at pharmacological concentrations (greater than 30 nM) when shunting of perfusate may have contributed to unperfused regions. 6. A network of mesenteric blood vessels estimated to contain approx. 6% vascular tissue by weight, with the remainder white fat cells, lymphatics and connective tissue, was also perfused. 7. Vasopressin (K0.5 = 0.3 nM) and noradrenaline (K0.5 = 30 nM) each increased oxygen uptake and perfusion pressure in a dose-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Flow-induced oxygen uptake by the perfused rat hindlimb is inhibited by vasodilators and augmented by norepinephrine: a possible role for the microvasculature in hindlimb thermogenesis.

Oxygen uptake in the perfused rat hindlimb was studied at 25 degrees C using an artificial perfusate, and the effects of perfusate flow rate, norepinephrine, and vasodilators were compared. Hindlimb oxygen uptake and perfusion pressure each increased as the flow rate was increased stepwise from 2 to 18.5 mL/min per hindlimb. At each flow rate, the rate of oxygen uptake was inhibited by the vasodilator nitroprusside (0.5 mM) and increased by norepinephrine (5 nM). A corresponding change in perfusion pressure also occurred, with norepinephrine leading to a marked increase and nitroprusside leading to a decrease; however, changes in oxygen uptake and pressure were not linearly related. The lactate/pyruvate ratio of the perfusate was used as an index of tissue perfusion and was determined at each flow rate. Lactate and pyruvate efflux increased as the flow rate was increased stepwise from 2 to 18.5 mL/min per hindlimb. At 2 mL/min per hindlimb, the lactate/pyruvate ratio was 15; at flow rates equal or greater than 4 mL/min per hindlimb, the ratio was constant at 9. Nitroprusside had no significant effect on the ratio at any flow rate even though a marked inhibitory effect on oxygen uptake was evident. Muscle content of high energy phosphates at 8 mL/min per hindlimb did not differ before and after treatment with vasodilators. In addition, the vasodilators had no apparent effect on skeletal muscle oxygen uptake or force development during electrical stimulation. The findings indicate that oxygen uptake by the hindlimb is not limited by inadequate perfusion and that oxygen uptake can be further increased by norepinephrine.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

PubMed1990

Capsaicin and dihydrocapsaicin stimulate oxygen consumption in the perfused rat hindlimb.

The capsaicinoid spice principles capsaicin and dihydrocapsaicin were shown to be thermogenic in the isolated rat hindlimb perfused with constant flow. Both principles elicited similar maximal increases in oxygen consumption (VO2) and perfusion pressure. For capsaicin, the mean maximal increase in VO2 was 1.39 +/- 0.28 mumol/g h or 23 per cent with a perfusion pressure increase of 15.8 +/- 2.8 mmHg, or 54.5 per cent. Dihydrocapsaicin increased VO2 by 1.13 +/- 0.24 or 20 per cent and a perfusion pressure rise of 14.2 +/- 5.0 mmHg or 49 per cent. Above 0.8 microM of either capsaicinoid there was inhibition of oxygen consumption after transient stimulation. Concurrent infusion of the adrenergic antagonists prazosin (alpha 1) and propranolol (beta) had little or no effect on the actions of either capsaicin or dihydrocapsaicin, nor did division of the somatic nerves to the hindlimb. These results indicate a local site of action of these principles in the hindlimb not mediated by a secondary release of catecholamines. Increases in both VO2 and pressure were significantly blocked by the vasodilator nitroprusside. This is in agreement with our previous findings that nitroprusside can block the effects of angiotensin, vasopressin and flow-induced increases in VO2 and perfusion pressure in the perfused hindlimb. The present findings suggest that capsaicin and dihydrocapsaicin can be thermogenic in the rat and that the mechanism of action directly involves vasoconstriction in some manner. We have previously suggested that there might be significant direct smooth muscle vascular consumption of oxygen during sustained vasoconstriction. The findings with capsaicin and dihydrocapsaicin are consistent with this hypothesis.

Animals↗

PubMed1989

Alpha 1-adrenergic control of contractility and coronary flow in the perfused rat heart.

Prazosin inhibition of phenylephrine plus propranolol (alpha 1-adrenergic agonist combination)-mediated inotropy, oxygen uptake, and coronary vasoconstriction of the perfused rat heart were compared with prazosin binding under identical conditions. Binding studies for the perfused heart indicated a population of high-affinity sites (Kd, 0.41 nM; Bmax, 13.2 pmol/g wet wt). Phenylephrine (50 microM) plus dl-propranolol (10 microM) did not significantly alter binding of 3.9 nM prazosin. The alpha 1-agonist combination mediated a dose-dependent increase in tension development and oxygen uptake and a decrease in coronary flow, each of which was inhibited by prazosin. The concentration of prazosin required for half-maximal inhibition of inotropy and oxygen uptake produced by 50-100 microM phenylephrine was 30-40 nM; no inhibition occurred at 2-3 nM prazosin concentration when binding sites were saturated. One nanomolar prazosin was required for half-maximal inhibition of the flow decrease produced by 10 microM phenylephrine. It is concluded that less than 1% of the alpha 1-binding sites are required for full development of inotropy and that this may result from an excess of binding sites or a small population of low-affinity receptors.

Animals↗

PubMed1988

Characterization of alpha 1-adrenergic receptors in perfused rat heart.

The characteristics of alpha 1-adrenergic receptors were investigated in perfused rat hearts at 37 degrees C. [3H]Prazosin was bound in a time-dependent manner and reached equilibrium at 15 min. Scatchard analysis of the specific binding isotherm for [3H]prazosin indicated a population of high affinity sites (Kd = 0.41 nM, Bmax = 13.2 pmol/g wet wt). Prazosin binding was displaced by epinephrine as well as by the adrenergic antagonists prazosin greater than phentolamine greater than yohimbine greater than propranolol. Specific prazosin binding was defined as that portion of the binding inhibited by 10 microM phentolamine; phentolamine and epinephrine displaced 3H-prazosin to the same level. [3H]Prazosin was not metabolized by the heart. When pre-labelled hearts were perfused at 37 degrees C with prazosin-free medium non-specific binding of [3H]prazosin decreased more rapidly (t0.5 = 4 min) than specific binding (t0.5 = 38 min). Perfusion of the heart at lower temperatures (less than 10 degrees C) decreased the rate of loss of nonspecific binding and prevented the loss of specific binding. Fractionation of [3H]prazosin perfused hearts at 0 degrees C, when dissociation was minimal, led to a loss of binding so that sarcolemma-enriched fractions contained approximately 2% of the binding sites present in the perfused heart. The binding characteristics of sarcolemma-enriched fractions (Kd 0.10 nM, Bmax 300 fmol/mg protein) differed significantly from those of the perfused heart. Exposure of the heart to 10 min of ischaemia prior to binding studies did not alter the characteristics of the [3H]prazosin binding sites. It is concluded that the perfused rat heart contains a population of alpha 1-adrenoceptors which differ from those of isolated sarcolemma preparations perhaps because of alterations that occur during sarcolemma isolation. The perfused heart should be an appropriate model system in which to study the relationship between receptor occupancy and biological response as well as the direct effects of perturbations such as ischaemia.

Animals↗

PubMed1988

Vasopressin and angiotensin II stimulate oxygen uptake in the perfused rat hindlimb.

Vasopressin and angiotensin II markedly stimulated oxygen uptake in the perfused rat hindlimb. The increase due to each agent approached 70% of the basal rate, and was greater than that produced by a maximal concentration of norepinephrine. Half-maximal stimulation occurred at 60 pM vasopressin, 0.5 nM angiotensin II and 10 nM norepinephrine. Angiotensins I and III were less potent than angiotensin II. For each agent, the dose-dependent increase in oxygen uptake coincided with a dose-dependent increase in perfusion pressure. The effects of both vasopressin and angiotensin to increase oxygen uptake and pressure were not inhibited by either phentolamine, propranolol or a combination of the two, but were completely inhibited by the vasodilator, nitroprusside. Nitroprusside also inhibited flow-induced increases in hindlimb oxygen uptake and perfusion pressure. The findings indicate a key role for the vascular system in the control of hindlimb oxygen uptake.

Angiotensin I↗

PubMed1986

Alpha-adrenergic receptors in rat skeletal muscle.

Sarcolemma-enriched preparations from muscles rich in slow oxidative red fibres contained specific binding sites for the alpha 1 antagonist, prazosin (e.g. soleus Kd 0.13 nM, Bmax 29 fmol/mg protein). Binding sites for prazosin were almost absent from white muscle. Displacement of prazosin binding from sarcolemma of soleus muscle (phentolamine greater than phenylephrine greater than idazoxan greater than yohimbine) suggested that the receptors were alpha 1. Binding sites for dihydroalprenolol (beta antagonist) were also more concentrated on red than white muscle and outnumbered prazosin sites by approx. 10:1. Binding sites for idazoxan (alpha 2 antagonist) were undetectable. Contamination of sarcolemma-enriched preparations by endothelial tissue indicated by the activity of angiotensin converting enzyme did not correlate with prazosin binding. It is concluded that post-synaptic alpha 1 adrenergic receptors are present on the sarcolemma of slow oxidative red fibres of rat skeletal muscle. The presence provides the mechanistic basis for apparent alpha-adrenergic effects to increase glucose and oxygen uptake in perfused rat hindquarter.

Animals↗

PubMed1984

Lysosomal thyroid hormone 5'-deiodinase.

Lysosomes prepared from rat liver and kidney after loading with the detergent Triton WR-1339 show membrane-bound 5'-deiodinase activity with marked specificity for 3,3',5'-triiodothyronine (reverse T3), lesser activity with respect to thyroxine (T4) and almost none towards 3,3',5'-triiodothyronine (T3). The enzyme is thiol dependent and shows maximal catalysis at pH 7.2. As many of the states known to alter thyroid hormone levels also affect lysosomal function, inhibition of the lysosomal 5'-deiodinase leading to an increase in intracellular reverse T3 may be an initiating mechanism for thyroid hormone change.

Animals↗

PubMedDate not supplied

Nonshivering thermogenesis in a marsupial (the tasmanian bettong Bettongia gaimardi) is not attributable to brown adipose tissue.

The Tasmanian bettong (Bettongia gaimardi, a marsupial) is a rat-kangaroo that increases nonshivering thermogenesis (NST) in response to norepinephrine (NE). This study attempted to assess whether brown adipose tissue (BAT), a specialized thermogenic effector, is involved in NST in the bettong. Regulatory NST, indicated by resting oxygen consumption (Vo2) of the whole body, was measured under conscious conditions at 20 degrees C with various stimuli: cold (4 degrees -5 degrees C) or warm (25 degrees C) acclimation, NE injection, and the beta3-adrenoceptor agonist (BRL) 37344. In line with the functional studies in vivo, the presence of BAT was evaluated by examining the expression of the uncoupling protein 1 (UCP1) with both rat cDNA and oligonucleotide probes. Both NE and BRL 37344 significantly stimulated NST in the bettong. After cold acclimation of the animals (at 4 degrees -5 degrees C for 2 wk), the resting Vo2 was increased by 15% and the thermogenic effect of NE was enhanced; warm-acclimated animals showed a slightly depressed response. However, no expression of UCP1 was detected in bettongs either before or after cold exposure (2 wk). These data suggest that the observed NST in the marsupial bettong is not attributable to BAT.

Adaptation, Physiological↗