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J H Coote

Publications and source records attributed to J H Coote.

At least 37 records · Page 2Linked to original sources
PubMed2001

Inhibitory effects of angiotensin II on barosensitive rostral ventrolateral medulla neurons of the rat.

1. The brain renin-angiotensin system can influence arterial baroreceptor reflex control of blood pressure (BP) through both direct and indirect effects on sympathetic premotor neurons of the rostral ventrolateral medulla (RVLM). The present study examined the direct effect of angiotensin (Ang) II applied by microiontophoresis on the ongoing activity of single RVLM neurons. 2. In 26 urethane-anaesthetized Wistar rats, recordings of single unit activities of barosensitive RVLM neurons were made from one barrel of a six-barrel micropipette assembly. The other five barrels were filled with either L-glutamate, AngII, valsartan (an AT1 receptor antagonist), PD 123177 (an AT2 receptor antagonist) and saline. All drugs were applied by microiontophoresis. 3. Mean BP was 83 +/- 3 mmHg. Application of AngII inhibited the ongoing activity of RVLM neurons, identified as barosensitive because their activity was inhibited by a phenylephrine- induced increase in BP, from 12.6 +/- 1.5 to 5.4 +/- 1.1 Hz (n=24; P < 0.001). Angiotensin II also inhibited the glutamate-evoked excitation of barosensitive RVLM neurons from 15 +/- 3 to 5.8 +/- 2.0 Hz (n=6; P < 0.001). Valsartan significantly increased neuronal activity from 9.5 +/- 2.3 to 13.5 +/- 3.2 Hz (n=7, P < 0.01), whereas PD 123177 significantly decreased neuronal activity from 13.5 +/- 3.5 to 9.9 +/- 2.8 Hz (n=13; P < 0.01). 4. The results suggest that AngII exerts a tonic inhibitory effect on barosensitive RVLM neurons, which is presumably mediated through AT1 receptor stimulation.

Adrenergic alpha-Agonists↗
PubMed2001

A novel and simple technique to allow detection of the position of the R-waves from intraventricular pressure waveforms: application to the conductance catheter method.

A simple and novel technique that utilizes the zero-crossing points of the first time derivative of intra-ventricular pressure (dP/dt) to mark systole, is proposed. Discrete differentiation of the sampled pressure waveform is calculated using a difference equation. Filtration of high-frequency noise in dP/dt is achieved using a low-pass Butterworth filter of order 4 and a cutoff frequency of 10 Hz. The filter is realized digitally using infinite impulse response filter stages. Double filtering of discrete dP/dt is used to eliminate time shifts. The methods are evaluated on data obtained from six large, white, anaesthetised and open chest pigs, instrumented with a conductance catheter. The zero-crossing points of the filtered dP/dt compare very well with the R-waves of the electrocardiogram (ECG) as markers of systole. The mean error is 1.3% of the duration of the heartcycle. Significantly, our results provide a solution to a problem often encountered with multiuse pressure-volume catheters when an ECG signal cannot be obtained. In this situation, the zero-crossing points of dP/dt, rather than the R-waves of the ECG, can be used as a marker of systole, thus enabling the construction of end-systolic pressure-volume relations to assess cardiac contractility.

Animals↗
PubMed2001

Tracing functionally identified neurones in a multisynaptic pathway in the hamster and rat using herpes simplex virus expressing green fluorescent protein.

Using a genetically modified herpes simplex virus encoding green fluorescent protein we sought to establish if this viral modification could be used in transneuronal tracing studies of the sympathetic nervous system. The herpes simplex virus encoding green fluorescent protein was injected into the adrenal medulla of three hamsters and six rats. After a suitable survival period, neurones in the sympathetic intermediolateral cell column of the thoracolumbar spinal cord, rostral ventral medulla and paraventricular nucleus of the hypothalamus were clearly identified by the presence of a green fluorescence in the cytoplasm of the neurones of both species. Thus, herpes simplex virus encoding green fluorescent protein labelled chains of sympathetic neurones in the hamster and rat and therefore has the potential to be used in transneuronal tracing studies of autonomic pathways in these species.

Animals↗
PubMed2001

Effects of direct sympathetic and vagus nerve stimulation on the physiology of the whole heart--a novel model of isolated Langendorff perfused rabbit heart with intact dual autonomic innervation.

A novel isolated Langendorff perfused rabbit heart preparation with intact dual autonomic innervation is described. This preparation allows the study of the effects of direct sympathetic and vagus nerve stimulation on the physiology of the whole heart. These hearts (n = 10) had baseline heart rates of 146 +/- 2 beats x min(-1) which could be increased to 240 +/- 11 beats x min(-1) by sympathetic stimulation (15 Hz) and decreased to 74 +/- 11 beats x min(-1) by stimulation of the vagus nerve (right vagus, 7 Hz). This model has the advantage of isolated preparations, with the absence of influence from circulating hormones and haemodynamic reflexes, and also that of in vivo preparations where direct nerve stimulation is possible without the need to use pharmacological agents. Data are presented characterising the preparation with respect to the effects of autonomic nerve stimulation on intrinsic heart rate and atrioventricular conduction at different stimulation frequencies. We show that stimulation of the right and left vagus nerve have differential effects on heart rate and atrioventricular conduction.

Adrenergic beta-Antagonists↗
PubMed2001

Effects of angiotensin II (AT1) receptor blockade on cardiac vagal control in heart failure.

The objective of the present study was to determine the autonomic effects of angiotensin II (AT(1)) receptor blocker therapy in heart failure. In a randomized double-blind cross-over study, we compared the effects of candesartan and placebo on baroreflex sensitivity and on heart rate variability at rest, during stress and during 24 h monitoring. Acute effects were assessed 4 h after oral candesartan (8 mg) and chronic effects after 4 weeks of treatment (dose titrated to 16 mg daily). The study group comprised 21 patients with heart failure [mean (S.E.M.) ejection fraction 33% (1%)], in the absence of angiotensin-converting enzyme (ACE) inhibitor therapy. We found that acute candesartan was not different from placebo in its effects on blood pressure or mean RR interval. Chronic candesartan significantly reduced blood pressure [placebo, 137 (3)/82 (3) mmHg; candesartan, 121 (4)/75 (2) mmHg; P<0.001; values are mean (S.E.M.)], but had no effect on mean RR interval [placebo, 857 (25) ms; candesartan, 857 (21) ms]. Compared with placebo there were no significant effects of acute or chronic candesartan on heart rate variability in the time domain and no consistent effects in the frequency domain. Baroreflex sensitivity assessed by the phenylephrine bolus method was significantly increased after chronic candesartan [placebo, 3.5 (0.5) ms/mmHg; candesartan, 4.8 (0.7) ms/mmHg; P<0.05], although there were no changes in cross-spectral baroreflex sensitivity. Thus, in contrast with previous results with ACE inhibitors, angiotensin II receptor blockade in heart failure did not increase heart rate variability, and there was no consistent effect on baroreflex sensitivity.

Aged↗
PubMed2000

Measurement of voltage-gated potassium currents in identified spinally-projecting sympathetic neurones of the paraventricular nucleus.

The paraventricular nucleus (PVN) of the hypothalamus modulates cardiovascular function via a sub-population of neurones which project directly to sympathetic centres of the spinal cord. Identification and patch-clamp recording from these neurones is difficult, however, because of the complex organisation and neuronal heterogeneity of the PVN. We report here on methods for the in vitro recording of voltage-gated potassium channel (K(V)) currents from those neurones within the PVN which project to the intermediolateral column of the rat spinal cord, and are believed to directly modulate cardiovascular function. We show K(V) channel currents of spinally projecting neurones to be slowly inactivating (tau >> 100 ms) and weakly sensitive to TEA (K(d)>10 mM). These methods will be useful for the study of K(V) and other ion channel modulation in spinally projecting neurones of the PVN.

Adrenergic Fibers↗
PubMed2000

Identification of branching paraventricular neurons of the hypothalamus that project to the rostroventrolateral medulla and spinal cord.

The paraventricular nucleus of the hypothalamus has efferent connections to autonomic nuclei known to ultimately regulate cardiovascular function. Studies have revealed projections to the sympathetic preganglionic neurons of the spinal cord and presympathetic motor neurons of the rostral ventrolateral medulla. This study set out to establish whether the same neurons in the paraventricular nucleus innervate both these regions. In rats the fluorescent neuroanatomical tracers FluoroGold, Fast Blue or Dextran tetramethyl rhodamine were injected into either the rostral ventrolateral medulla or T2 region of the spinal cord. After a suitable survival period (five to seven days) three populations of neurons could be identified in the paraventricular nucleus, double-labelled neurons and single-labelled neurons resulting from the injections into the spinal cord or injections into the rostral ventrolateral medulla. The neurons were of similar size regardless of the dye content. Most neurons were found in the parvocellular subdivision of the mid rostral paraventricular nucleus. The number of labelled neurons decreased in the caudal sections. This study provides an anatomical basis for three means of influence that the paraventricular nucleus can have on sympathetic activity; a hierarchical in series projection via the rostral ventrolateral medulla; a projection running in parallel with this but bypassing the rostroventrolateral medulla; and a branching population innervating neurons in both the rostral ventrolateral medulla and spinal cord. The paraventricular nucleus of the hypothalamus is an important brain area concerned with maintaining cardiovascular homeostasis. This anatomical study has not only provided confirmatory evidence that direct projections arising from the paraventricular hypothalamic nucleus do project to the rostral ventrolateral medulla and spinal cord, regions known to influence cardiovascular regulation. The study has identified a branching projection originating in the paraventricular nucleus of the hypothalamus that projects to both the rostral ventrolateral medulla and the spinal cord. Thus the paraventricular nucleus of the hypothalamus has three pathways in which to influence cardiovascular homeostasis.

Amidines↗
PubMed2000

Leaky scanning is the predominant mechanism for translation of human papillomavirus type 16 E7 oncoprotein from E6/E7 bicistronic mRNA.

Human papillomaviruses (HPV) are unique in that they generate mRNAs that apparently can express multiple proteins from tandemly arranged open reading frames. The mechanisms by which this is achieved are uncertain and are at odds with the basic predictions of the scanning model for translation initiation. We investigated the unorthodox mechanism by which the E6 and E7 oncoproteins from human papillomavirus type 16 (HPV-16) can be translated from a single, bicistronic mRNA. The short E6 5' untranslated region (UTR) was shown to promote translation as efficiently as a UTR from Xenopus beta-globin. Insertion of a secondary structural element into the UTR inhibited both E6 and E7 expression, suggesting that E7 expression depends on ribosomal scanning from the 5' end of the mRNA. E7 translation was found to be cap dependent, but E6 was more dependent on capping and eIF4F activity than E7. Insertion of secondary structural elements at various points in the region upstream of E7 profoundly inhibited translation, indicating that scanning was probably continuous. Insertion of the E6 region between Renilla and firefly luciferase genes revealed little or no internal ribosomal entry site activity. However when E6 was located at the 5' end of the mRNA, it permitted over 100-fold-higher levels of downstream cistron translation than did the Renilla open reading frame. Internal AUGs in the E6 region with strong or intermediate Kozak sequence contexts were unable to inhibit E7 translation, but initiation at the E7 AUG was efficient and accurate. These data support a model in which E7 translation is facilitated by an extreme degree of leaky scanning, requiring the negotiation of 13 upstream AUGs. Ribosomal initiation complexes which fail to initiate at the E6 start codon can scan through to the E7 AUG without initiating translation, but competence to initiate is achieved once the E7 AUG is reached. These findings suggest that the E6 region of HPV-16 comprises features that sponsor both translation of the E6 protein and enhancement of translation at a downstream site.

5' Untranslated Regions↗
PubMed2000

Nitric oxide and cardiac autonomic control in humans.

Cardiac autonomic control is of prognostic significance in cardiac disease, yet the control mechanisms of this system remain poorly defined. Animal data suggest that nitric oxide (NO) modulates cardiac autonomic control. We investigated the influence of NO on the baroreflex control of heart rate in healthy human subjects. In 26 healthy male volunteers (mean age, 23+/-5 years), we measured heart rate variability and baroreflex sensitivity during inhibition of endogenous NO production with N(G)-monomethyl-L-arginine (L-NMMA) (3 mg/kg per hour) and during exogenous NO donation with sodium nitroprusside (1 to 3 mg/h). Increases from baseline (Delta) in high-frequency (HF) indexes of heart rate variability were smaller with L-NMMA in comparison to an equipressor dose of the control vasoconstrictor phenylephrine (12 to 42 microg/kg per hour): Deltaroot mean square of successive RR interval differences (DeltaRMSSD)=23+/-32 versus 51+/-48 ms (P<0.002); Deltapercentage of successive RR interval differences >50 ms (DeltapNN50)=5+/-15% versus 14+/-12% (P<0.05); and DeltaHF normalized power=-2+/-7 versus 9+/-8 normalized units (P<0.01), respectively. Relative preservation of these indexes was observed during unloading of the baroreflex with sodium nitroprusside compared with a matched fall in blood pressure produced by a control vasodilator, hydralazine (9 to 18 mg/h): DeltaRMSSD=-8+/-8 versus -24+/-15 ms (P<0.001); DeltapNN50=-6+/-11% versus -15+/-19% (P<0.01); DeltaHF normalized power=-7+/-13 versus -13+/-11 normalized units (P<0.05), respectively. The change in cross-spectral alpha-index calculated as the square root of the ratio of RR interval power to systolic spectral power in the HF band (although not alpha-index calculated in the same way for the low-frequency bands or baroreflex sensitivity assessed by the phenylephrine bolus method) was attenuated with L-NMMA compared with phenylephrine (Delta=4+/-8 versus 14+/-15 ms/mm Hg, respectively; P<0.02) and with sodium nitroprusside compared with hydralazine (Delta=-7+/-6 and -9+/-7 ms/mm Hg, respectively; P<0.05). In conclusion, these data demonstrate that NO augments cardiac vagal control in humans.

Adolescent↗
PubMed2000

Paraventricular neurones elicit a volume expansion-like change of activity in sympathetic nerves to the heart and kidney in the rabbit.

The role of the paraventricular nucleus (PVN) of the hypothalamus in the cardiovascular response induced by blood volume expansion was examined in anaesthetised rabbits, in which simultaneous recordings were made from a renal sympathetic nerve and one of other sympathetic nerves, the inferior cardiac nerve, a splanchnic nerve and an adrenal nerve. Activation of PVN neurones, by discrete injections (25-100 nl) of d,l-homocysteic acid (DLH, 0.2 M) produced one main pattern of sympathetic nerve activity accompanying a pressor response (57-86 % of PVN sites). This was a decrease in renal sympathetic activity (27 +/- 12 %) and an increase in splanchnic (60 +/- 12 %), adrenal (31 +/- 9 %) and cardiac (42 +/- 8 %) sympathetic activity. Sites in the PVN from which these combinations of nerve activity were obtained were not confined to a specific subnucleus. An increase in renal sympathetic activity which was reversed to a decrease by reducing the volume of DLH injected was obtained at 10 sites in the PVN. These sites were mainly located in the dorsal parvocellular subnucleus. Varying combinations of sympathetic activation were obtained at a minority of sites. It is concluded that the PVN can evoke a differential pattern of sympathetic discharge which may be functionally significant in the control of blood volume regulation, as it mimics that seen on acute volume expansion.

Adrenal Glands↗
PubMed1999

Effect of sympathectomy on the expression of NMDA receptors in the spinal cord.

The expression of NMDA receptors in the intermediolateral (IML) region of the upper thoracic spinal cord, was studied in 3 week old rats. The effect of section of the cervical sympathetic nerve on neuronal cell number and receptor expression was examined up to two weeks after the operation. Age-matched sham-operated and unoperated animals were used as controls. It was shown using quantitative autoradiography with the NMDA receptor antagonist [(3)H]MK-801 (dizocilpine maleate), that there was a marked downregulation of receptors in all groups of animals, beginning at approximately 4 weeks of age. However after sympathectomy, which resulted in the death of 44% of neurones in the IML by 7 days, there was a significant increase in receptor density per neurone compared to sham-operated controls. In the control animals there was a significant increase in the Kd value of the binding between 21 and 24 days after birth indicating an increased expression of a low affinity receptor, but no such increase was seen after axotomy. The results are consistent with two populations of NMDA receptors being transiently expressed in the IML in developing animals, and the higher affinity receptor being down-regulated between 4 and 5 weeks of age. The presence of the high affinity receptor subtype may predispose neurones to die after axotomy.

Animals↗
PubMed1999

Fast excitatory post synaptic potentials and their response to catecholaminergic antagonists in rat sympathetic preganglionic neurones in vitro.

In an in vitro slice preparation from neonatal rats intracellular recordings were made from electrophysiologically identified sympathetic preganglionic neurones. Electrical stimulation in the lateral funiculus (>500 microm) from the recording site elicited a mono- or polysynaptic excitatory post synaptic potential. The latter potential was blocked with the dopamine D2 antagonist haloperidol but not with the dopamine D1 antagonist SCH 23390. We therefore report the first showing of a functional descending pathway in an in vitro slice preparation describing both the transmitter and the receptor subtype involved and physiologically show that dopamine may exert an indirect excitatory influence on sympathetic preganglionic neurones possibly via interneurones present in the spinal cord.

Autonomic Fibers, Preganglionic↗
PubMed1999

Inhibitory and indirect excitatory effects of dopamine on sympathetic preganglionic neurones in the neonatal rat spinal cord in vitro.

Regions of the thoraco-lumbar spinal cord containing sympathetic preganglionic neurones are rich in dopamine terminals. To determine the influence of this innervation intracellular recordings were made from antidromically identified sympathetic preganglionic neurones in (400 micrometers) transverse neonatal rat spinal cord slices. Dopamine applied by superfusion caused a slow monophasic hyperpolarisation in 46% of sympathetic preganglionic neurones, a slow monophasic depolarisation in 28% of sympathetic preganglionic neurones and a biphasic effect consisting of a slow depolarisation followed by a slow hyperpolarisation or vice-versa in 23% of sympathetic preganglionic neurones. Three percent of sympathetic preganglionic neurones did not respond to the application of dopamine. Low Ca2+/high Mg2+ Krebs solution or TTX did not change the resting membrane potential but abolished the slow depolarisation elicited by dopamine, indicating this was synaptic and did not prevent the dopamine induced hyperpolarisation. The dopamine induced slow hyperpolarisation was mimicked by the selective D1 agonists SKF 38393 or SKF 81297-C and blocked by superfusion with the D1 antagonist SCH 23390. It was not prevented by superfusion of the slices with alpha1 or alpha2 or beta-adrenoceptor antagonists, whereas the inhibitory or excitatory actions of adrenaline were prevented by alpha1 or alpha2 antagonists, respectively. The dopamine induced slow depolarisation occurring in a sub-population of sympathetic preganglionic neurones was mimicked by quinpirole, a D2 agonist, and blocked by haloperidol, a D2 antagonist. Haloperidol did not block the dopamine induced hyperpolarisations. Dopamine also induced fast synaptic activity which was mimicked by a D2 agonist and blocked by haloperidol. D1 agonists did not elicit fast synaptic activity.

Adrenergic Antagonists↗
PubMed1999

Terminals of paraventricular spinal neurones are closely associated with adrenal medullary sympathetic preganglionic neurones: immunocytochemical evidence for vasopressin as a possible neurotransmitter in this pathway.

A recent study using transsynaptically transported pseudorabies virus, injected into the adrenal gland, showed labelled neurones in the paraventricular nucleus (PVN) of the hypothalamus, indicating that these neurones send projections to sympathoadrenal preganglionic neurones (SPNs). However, this technique cannot conclusively demonstrate that the pathway is monosynaptic. In order to investigate the possibility of a direct projection from the PVN to SPNs, the present study used the anterograde tracer biotin dextran amine to label paraventricular spinal projections and the retrograde tracer cholera toxin B conjugated to horseradish peroxidase to label SPNs. In addition, because electrophysiological evidence suggests vasopressin to be a neurotransmitter candidate in this pathway, immunocytochemical identification of the peptide and retrograde labelling of SPNs to the adrenal medulla were used to investigate this. The results of these studies show spinally projecting paraventricular axons with terminal varicosities closely associated with SPNs. Therefore some of these associations may represent boutons forming synaptic contact on SPNs. Similarly, vasopressin fibres were found close to the dendrites and soma of SPNs. It is suggested that spinal axons originating from paraventricular neurones can provide a direct influence on adrenal medullary function, that vasopressin is a possible neurotransmitter involved in some of these connections and this is one means by which the paraventricular nucleus can generate a defence to stressful stimuli.

Adrenal Medulla↗
PubMed1999

D(1)-like dopamine receptors on retrogradely labelled sympathoadrenal neurones in the thoracic spinal cord of the rat.

Cellular localization of dopamine D(1)-like receptors was accomplished on target-specified sympathoadrenal preganglionic neurones using the radioligand [(3)H]SCH23390. Sympathoadrenal neurones were retrogradely labelled with cholera B subunit conjugated to horseradish peroxidase and were detected in segments T(1) to T(13) with a predominance at T(8)/T(9). Binding of the selective D(1)-like radioligand [(3)H]SCH23390 was associated with the retrogradely labelled sympathoadrenal neurones in longitudinal/horizontal sections of thoracic spinal cord. D(1)-like receptor localization on target-specific neurones was determined in more than half of the spinal cord sections and was associated predominantly with the cell soma and principal proximal dendrites in the intermediolateral cell column of the spinal grey matter. D(2)-like receptor localization was not associated with retrogradely labelled sympathoadrenal neurones but a higher degree of specific binding was noted in more medial aspects of the spinal grey matter. This is the first successful demonstration of receptor localization combining two quite different techniques and provides conclusive anatomical evidence for D(1)-like receptor localization on sympathetic preganglionic neurones that project to the adrenal medulla.

Adrenal Medulla↗
PubMed1999

The influence of the paraventricular nucleus on baroreceptor dependent caudal ventrolateral medullary neurones of the rat.

The question of whether neurones in the paraventricular nucleus (PVN) of the hypothalamus have an influence on barosensitive vasomotor neurones of the caudal ventrolateral medulla (CVL) was addressed in this study using anaesthetised rats. Extracellular microelectrode recordings were made from 27 vasomotor neurones in the CVL, identified by their cardiac cycle-related probability of discharge, by the increase in activity in response to an increase in arterial blood pressure produced by i.v. phenylephrine and by the decrease in activity in response to a decrease in blood pressure produced by i.v. nitroprusside. Activation of neurones at different sites in the PVN with a microinjection of d,l-homocysteic acid (DLH) activated 12 CVL neurones and inhibited 6 CVL neurones. In four CVL units single-shock electrical stimulation at a PVN pressor site, first identified with DLH, elicited a synaptic action potential with a latency of 29+/-0.3 ms. It is concluded that PVN neurones can influence the CVL vasomotor neurones directly. This could be one means by which PVN-depressor and sympatho-inhibitory effects are produced.

Anesthesia↗