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J A Coles

Publications and source records attributed to J A Coles.

At least 19 recordsLinked to original sources
PubMed2001

Mapping extracellular pH in rat brain gliomas in vivo by 1H magnetic resonance spectroscopic imaging: comparison with maps of metabolites.

The value of extracellular pH (pH(e)) in tumors is an important factor in prognosisand choice of therapy. We demonstrate here that pH(e) can be mappedin vivo in a rat brain glioma by (1)H magnetic resonance spectroscopic imaging (SI) of the pH buffer (+/-)2-imidazole-1-yl-3-ethoxycarbonylpropionic acid (IEPA). (1)H SI also allowed us to map metabolites, and, to better understand the determinants of pH(e), we compared maps of pH(e), metabolites, and the distribution of the contrast agent gadolinium1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraaceticacid (Gd-DOTA). C6 cells injected in caudate nuclei of four Wistar rats gave rise to gliomas of approximately 10 mm in diameter. Three mmols of IEPA were injected in the right jugular vein from t = 0 to t = 60 min. From t = 50 min to t = 90 min, spin-echo (1)H SI was performed with an echo time of 40 ms in a 2.5-mm slice including the glioma (nominal voxel size, 2.2 microl). IEPA resonances were detected only within the glioma and were intense enough for pH(e) to be calculated from the chemical shift of the H2 resonance in almost all voxels of the glioma. (1)H spectroscopic images with an echo time of 136 ms were then acquired to map metabolites: lactate, choline-containing compounds (tCho), phosphocreatine/creatine, and N-acetylaspartate. Finally, T(1)-weighted imaging after injection of a bolus of Gd-DOTA gave a map indicative of extravasation. On average, the gradient of pH(e) (measured where sufficient IEPA was present) from the center to the periphery was not statistically significant. Mean pH(e) was calculated for each of the four gliomas, and the average was 7.084 +/- 0.017 (+/- SE; n = 4 rats), which is acid with respect to pH(e) of normal tissue. After normalization of spectra to their water peak, voxel-by-voxel comparisons of peak areas showed that N-acetylaspartate, a marker of neurons, correlated negatively with IEPA (P < 0.0001) and lactate (P < 0.05), as expected of a glioma surrounded by normal tissue. tCho (which may indicate proliferation) correlated positively with pH(e) (P < 0.0001). Lactate correlated positively with tCho (P < 0.0001), phosphocreatine/creatine (P < 0.001), and Gd-DOTA (P < 0.0001). Although lactate is exported from cells in association with protons, within the gliomas, no evidence was observed that pH(e) was significantly lower where lactate concentration was higher. These results suggest that lactate is produced mainly in viable, well-perfused, tumoral tissue from which proton equivalents are rapidly cleared.

Animals↗
PubMed2001

Opioid preconditioning: myocardial function and energy metabolism.

BACKGROUND: Opioid receptor agonists are involved in ischemic preconditioning and natural hibernation. The aim of this study was to determine whether pretreatment with D-Ala2-Leu5-enkephalin or morphine confers cardioprotection in large mammalian hearts. We assessed myocardial functional recovery and global energy metabolism after ischemic cold storage. METHODS: After pretreatment with D-Ala2-Leu5-enkephalin, morphine sulfate, or saline (n = 6 each), swine hearts were excised and stored for 75 minutes at 4 degrees C, then reperfused in a four-chamber isolated working heart apparatus. Serial myocardial biopsies were performed to assess cellular energy metabolism. RESULTS: Improved systolic (cardiac output, contractility) and diastolic (tau) left ventricular functions were observed in hearts pretreated with D-Ala2-Leu5-enkephalin or morphine. These benefits were not correlated with changes in high-energy phosphate levels. Cardiac enzyme leakage (creatine kinase, troponin-I) was similar among treated and control groups. Lactate efflux increased significantly in controls, but not in opioid-pretreated hearts (p < 0.01) at 75 minutes of reperfusion. CONCLUSIONS: D-Ala2-Leu5-enkephalin and morphine pretreatments improve postischemic function after cold storage of swine hearts. Postischemic lactate reduction, but not high-energy phosphate levels, may account for the observed cardioprotective effects.

Animals↗
PubMed2001

Ammonium in nervous tissue: transport across cell membranes, fluxes from neurons to glial cells, and role in signalling.

Most, but not all, animal cell membranes are permeable to NH3, the neutral, minority form of ammonium which is in equilibrium with the charged majority form NH4+. NH4+ crosses many cell membranes via ion channels or on membrane transporters, and cultured mammalian astrocytes and glial cells of bee retina take up NH4+ avidly, in the latter case on a Cl(-)-cotransporter selective for NH4+ over K+. In bee retina, a flux of ammonium from neurons to glial cells is an essential component of energy metabolism, which involves a flux of alanine from glial cells to neurons. In mammalian brain, both glutamate and ammonium are taken up preferentially by astrocytes and form glutamine. Glutamine is transferred to neurons where it is deamidated to re-form glutamate; the maintenance of this cycle appears to require a substantial flux of ammonium from neurons to astrocytes. In addition to maintaining the glial cell content of fixed N (a "bookkeeping" function), ammonium is expected to participate in the regulation of glial cell metabolism (a signalling function): it will increase conversion of glutamate to glutamine, and, by activating phosphofructokinase and inhibiting the alpha-ketoglutarate dehydrogenase complex, it will tend to increase the formation of lactate.

Animals↗
PubMed2000

Spontaneous neuronal activity in organotypic cultures of mouse dorsal root ganglion leads to upregulation of calcium channel expression on remote Schwann cells.

It is well established that neurons regulate the properties of both central and peripheral glial cells. Some of these neuro-glial interactions are modulated by the pattern of neuronal electrical activity. In the present work, we asked whether blocking the electrical activity of dorsal root ganglion (DRG) neurons in vitro by a chronic treatment with tetrodotoxin (TTX) would modulate the expression of the T-type Ca(2+) channel by mouse Schwann cells. When recorded in their culture medium, about one-half of the DRG neurons spontaneously fired action potentials (APs). Treatment for 4 days with 1 microM TTX abolished both spontaneous and evoked APs in DRG neurons and in parallel significantly reduced the percentage of Schwann cells expressing Ca(2+) channel currents. On the fraction of Schwann cells still expressing Ca(2+) channel currents, these currents had electrophysiological parameters (mean amplitude, mean inactivation time constant, steady-state inactivation curve) similar to those of control cultures. Co-treatment for 4 days with 1 microM TTX and 2 mM CPT-cAMP, a cAMP analogue that induces the expression de novo of Ca(2+) channel currents in Schwann cells deprived of neurons, maintained the percentage of Schwann cells expressing Ca(2+) channel currents, showing that TTX does not directly affect the expression of Ca(2+) channel currents by Schwann cell. We conclude that blocking spontaneous activity of DRG neurons in vitro downregulates Ca(2+) channel expression by Schwann cells. These results strongly suggest that DRG neurons upregulate Ca(2+) channel expression by Schwann cells via the release of a diffusible factor whose secretion is dependent on electrical activity.

Animals↗
PubMed2000

A Cl(-) cotransporter selective for NH(4)(+) over K(+) in glial cells of bee retina.

There appears to be a flux of ammonium (NH(4)(+)/NH(3)) from neurons to glial cells in most nervous tissues. In bee retinal glial cells, NH(4)(+)/NH(3) uptake is at least partly by chloride-dependant transport of the ionic form NH(4)(+). Transmembrane transport of NH(4)(+) has been described previously on transporters on which NH(4)(+) replaces K(+), or, more rarely, Na(+) or H(+), but no transport system in animal cells has been shown to be selective for NH(4)(+) over these other ions. To see if the NH(4)(+)-Cl(-) cotransporter on bee retinal glial cells is selective for NH(4)(+) over K(+) we measured ammonium-induced changes in intracellular pH (pH(i)) in isolated bundles of glial cells using a fluorescent indicator. These changes in pH(i) result from transmembrane fluxes not only of NH(4)(+), but also of NH(3). To estimate transmembrane fluxes of NH(4)(+), it was necessary to measure several parameters. Intracellular pH buffering power was found to be 12 mM. Regulatory mechanisms tended to restore intracellular [H(+)] after its displacement with a time constant of 3 min. Membrane permeability to NH(3) was 13 microm s(-1). A numerical model was used to deduce the NH(4)(+) flux through the transporter that would account for the pH(i) changes induced by a 30-s application of ammonium. This flux saturated with increasing [NH(4)(+)](o); the relation was fitted with a Michaelis-Menten equation with K(m) approximately 7 mM. The inhibition of NH(4)(+) flux by extracellular K(+) appeared to be competitive, with an apparent K(i) of approximately 15 mM. A simple standard model of the transport process satisfactorily described the pH(i) changes caused by various experimental manipulations when the transporter bound NH(4)(+) with greater affinity than K(+). We conclude that this transporter is functionally selective for NH(4)(+) over K(+) and that the transporter molecule probably has a greater affinity for NH(4)(+) than for K(+).

Acid-Base Equilibrium↗
PubMed1999

A rapid wavelength changer based on liquid crystal shutters for use in ratiometric microspectrophotometry.

Many of the fluorescent indicator molecules most useful for measuring intracellular concentrations of ions of biological importance, such as Ca2+ or H+, require illumination first at one wavelength, at which the fluorescence depends strongly on the concentration of the ion, and then at another wavelength (e.g. the isosbestic point), so that a ratio can be obtained. Existing wavelength changers are mechanical and involve moving filters, mirrors or gratings. These systems have the disadvantage that they introduce mechanical shocks that can interfere with simultaneous electrophysiological recording. In addition, they require special electrical driving systems and are relatively expensive, especially if they are capable of switching rapidly. We describe a new wavelength changer based on liquid crystal shutters which has the following advantages: (1) it has no mechanical moving parts; (2) it can switch rapidly (@1 ms) and in any desired pattern (off - on1 - off - on2 - off, etc.); (3) it is driven by a low-power 15-V pulse; and (4) it is substantially cheaper than existing wavelength changers. Its limitations are that it does not pass wavelengths shorter than about 400 nm and transmission in the range 430-700 nm is only 20-40%.

Calcium↗
PubMed1999

Chloride-dependent transport of NH4+ into bee retinal glial cells.

Mammalian astrocytes convert glutamate to glutamine and bee retinal glial cells convert pyruvate to alanine. To maintain such amination reactions these glial cells may take up NH4+/NH3. We have studied the entry of NH4+/NH3 into bundles of glial cells isolated from bee retina by using the fluorescent dye BCECF to measure pH. Ammonium caused intracellular pH to decrease by a saturable process: the rate of change of pH was maximal for an ammonium concentration of about 5 mM. This acidifying response to ammonium was abolished by the loop diuretic bumetanide (100 microM) and by removal of extracellular Cl-. These results strongly suggest that ammonium enters the cell by contransport of NH4+ with Cl-. Removal of extracellular Na+ did not abolish the NH(4+)-induced acidification. The NH(4+)-induced pH change was unaffected when nearly all K+ conductance was blocked with 5 mM Ba2+ showing that NH4+ did not enter through Ba(2+)-sensitive ion channels. Application of 2 mM NH4+ led to a large increase in total intracellular proton concentration estimated to exceed 13.5 mEq/L. As the cell membrane appeared to be permeable to NH3, we suggest that when NH4+ entered the cells, NH3 left, so that protons were shuttled into the cell. This shuttle, which was strongly dependent on internal and external pH, was quantitatively modelled. In retinal slices, 2 mM NH4+ alkalinized the extracellular space: this alkalinization was reduced in the absence of bath Cl-. We conclude that NH4+ enters the glial cells in bee retina on a cotransporter with functional similarities to the NH4+(K+)-Cl- cotransporter described in kidney cells.

Animals↗
PubMed1998

Functional Ca2+ and Na+ channels on mouse Schwann cells cultured in serum-free medium: regulation by a diffusible factor from neurons and by cAMP.

Regulation of expression of functional voltage-gated ion channels for inward currents was studied in Schwann cells in organotypic cultures of dorsal root ganglia from E19 mouse embryos maintained in serum-free medium. Of the Schwann cells that did not contact axons, 46.5% expressed T-type Ca2+ conductances (ICaT). Two days or more after excision of the ganglia, and consequent disappearance of neurites, ICaT were detectable in only 10.9% of the cells, and the marker 04 disappeared. On Schwann cells deprived of neurons, T- (but not L-) type Ca2+ conductances were re-induced by weakly hydrolysable analogues of cAMP, and by forskolin (an activator of adenylyl cyclase) after long-term treatment (4 days). With CPT cAMP (0.1-2 mM), 8Br cAMP, db cAMP or forskolin (0.01 or 0.1 mM), the proportion of cells with ICaT was not significantly different from the proportion in the cultures with neurons. These agents also induced expression in some cells of tetrodotoxin-resistant Na+ currents, which were rarely induced by neurons, but 04 was not re-induced by cAMP analogue treatments that re-induced ICaT. Inward currents (Ba2+ or Na+) were partly restored (P < 0.05) on Schwann cells cultured for 6-7 days beneath a filter bearing cultured neurons. In contrast, addition of neuron-conditioned medium was ineffective. The results suggest that neurons activate, via diffusible and degradable factors, a subset of Schwann cell cAMP pathways leading to expression of IcaT, and activate additional non-cAMP pathways that lead to expression of 04.

Animals↗
PubMed1998

Lactate is released and taken up by isolated rabbit vagus nerve during aerobic metabolism.

To determine if lactate is produced during aerobic metabolism in peripheral nerve, we incubated pieces of rabbit vagus nerve in oxygenated solution containing D-[U-14C]glucose while stimulating electrically. After 30 min, nearly all the radioactivity in metabolites in the nerve was in lactate, glucose 6-phosphate, glutamate, and aspartate. Much lactate was released to the bath: 8.2 pmol (microg dry wt)(-1) from the exogenous glucose and 14.2 pmol (microg dry wt)(-1) from endogenous substrates. Lactate release was not increased when bath PO2 was decreased, indicating that it did not come from anoxic tissue. When the bath contained [U-14C]lactate at a total concentration of 2.13 mM and 1 mM glucose, 14C was incorporated in CO2 and glutamate. The initial rate of formation of CO2 from bath lactate was more rapid than its formation from bath glucose. The results are most readily explained by the hypothesis that has been proposed for brain tissue in which glial cells supply lactate to neurons.

Aerobiosis↗
PubMed1997

Selective downregulation of an inactivating K+ conductance by analogues of cAMP in mouse Schwann cells.

1. Voltage-dependent K+ conductances on Schwann cells in organotypic cultures of mouse dorsal root ganglia were classified as inactivating or sustained (responsible for currents IA and IK, respectively). IA is known to be much reduced on Schwann cells that contact neurites. 2. In the absence of neurones, IA and IK were present. IA, but not IK, was markedly reduced (by 80% after 105 h of treatment) by 2 mM 8-(4-chlorophenylthio)-cAMP (cpt-cAMP), a weakly hydrolysable analogue of cAMP. The effect did not appear for at least 2 h and was maximal after about 100 h. 3. The effect of 1 mM cpt-cAMP was abolished in the presence of an inhibitor of protein kinases, N-[2-bromocinnamyl(amino)ethyl]-5-isoquinolinesulphonamide (H-89, 10 microM). 4. Other analogues of cAMP, but not an analogue of cGMP (8-bromo-cGMP), also reduced IA. 5. We conclude that IA, but not IK, can be downregulated by activation of the protein kinase A pathway.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1997

Potassium homeostasis and glial energy metabolism.

Since capillaries appear not to contribute significantly to rapid removal of K+ from brain tissue, the K+ released into extracellular clefts by neurons at the onset of electrical activity is presumably removed either by redistribution in the clefts or by uptake into cells. What appear to be the three major processes require no energy from the glial cells. These are diffusion through the extracellular clefts, spatial buffering by glial cells, and net uptake of K+ into glial cells through glial K+ channels associated with uptake of Cl- through an independent Cl- conductance. There is a relatively slow uptake by the Na+/K+-ATPase, which directly consumes ATP. In addition, some glial cells take up K+ on the Na+/K+/2Cl- cotransporter, which leads indirectly to energy consumption when the Na+ is subsequently pumped out. Currently available data suggest that the glial energy metabolism devoted to K+ homeostasis is less than a tenth of the total tissue energy metabolism, even under conditions of pathologically high extracellular [K+]. Hence, in situ, it is possible that glial cells could function with much less ATP than neurons do. All the various routes of muffling of changes in extracellular [K+] can be modulated, directly or indirectly, by transmitters liberated by neurons. A consequence of this could be regulation of the entry of Na+ into glial cells such that the Na+/K+-ATPase is activated. The degree of activation might be adjusted so that the resulting activation of the glial glycolytic pathway is appropriate to the provision of the quantity of metabolic substrates required by the neurons.

Animals↗
PubMed1997

The separation and characterisation of haemocytes from the mussel Mytilus edulis.

The separation of haemocytes from the mussel Mytilus edulis was carried out on continuous Percoll gradients. The haemocytes separated into three distinct layers, the first comprised 97% basophilic cells, the third comprised 84% eosinophilic cells and the middle layer was a mixture of eosinophilic and basophilic cells. Enzyme cytochemistry demonstrated arylsulphatase, phenol oxidase and peroxidase associated with the haemocytes from the third layer. Lectin-binding studies showed differential binding of lectins to the separated cells. The ultrastructural morphology demonstrated that the first layer of cells was composed predominantly of small agranular cells with a high nucleus to cytoplasm ratio. The second layer comprised a mixture of cells with the majority being granular cells with small granules. The third layer was almost exclusively composed of granular cells with small and large granules. Assays to assess the function of the different cells demonstrated that respiratory burst activity, measured as the reduction of cytochrome-c, was carried out almost entirely by the eosinophilic haemocytes. Similarly, levels of phagocytosis, measured as uptake of Escherichia coli, were much higher in the eosinophilic haemocytes. Of the potential mitogenic factors investigated, concanavalin A and pokeweed mitogen showed some evidence of inducing haemocyte proliferation.

Animals↗
PubMed1996

Effects of photoreceptor metabolism on interstitial and glial cell pH in bee retina: evidence of a role for NH4+.

1. Measurements were made with pH microelectrodes in superfused slices of the retina of the honey-bee drone. In the dark, the mean +/- S.E.M. pH values in the three compartments of the tissue were: neurones (photoreceptors), 6.99 +/- 0.04; glial cells (outer pigment cells), 7.31 +/- 0.03; extracellular space, 6.60 +/- 0.03. 2. Stimulation of the photoreceptors with light caused transient pH changes: a decrease in the photoreceptors (pHn) and in the glial cells (pHg), and an increase in the interstitial clefts (pHo). 3. The effects of inhibition and activation of aerobic metabolism showed that part, perhaps all, of the light-induced delta pHo resulted from the increased aerobic metabolism in the photoreceptors. 4. Addition of 2 mM NH4+ to the superfusate produced changes in pHo and pHg of the same sign as and similar amplitude to those caused by light stimulation. Manipulation of transmembrane pH gradients had similar effects on changes in pHo induced by light or by exogenous NH4+. 5. Measurements with NH(4+)-sensitive microelectrodes showed that stimulation of aerobic metabolism in the photoreceptors increased [NH4+]o and also that exogenous NH4+/NH3 was taken up by cells, presumably the glial cells. 6. We conclude that within seconds of an increase in the aerobic metabolism in the photoreceptors, they release an increased amount of NH4+/NH3 which affects pHo and enters glial cells. Other evidence suggests that in drone retina the glial cells supply the neurones with amino acids as substrates of energy metabolism; the present results suggest that fixed nitrogen is returned to the glial cells as NH4+/NH3.

2,4-Dinitrophenol↗
PubMed1996

Signalling from neurones to glial cells in invertebrates.

Recent work shows that glial cells in species throughout the animal kingdom appear to contribute to the functioning of the neurones and are equipped to receive signals from them. However, the detailed mechanisms of the signalling and its role in vivo are generally unclear. Parts of some invertebrate nervous systems are particularly favourable for addressing these problems, and the four preparations that have been studied most intensively are the subject of this review. Between the giant axons and their Schwann glial cells in squid and crayfish, within snail brain, and in leech ganglion, there appear to be multiple, and in some cases very complex, signalling pathways, whose precise functions remain to be elucidated. In bee retina only a single signal to the glia has been demonstrated, and its function appears to be to activate transfer of metabolic substrates to the photoreceptor neurones.

Animals↗
PubMed1995

Conditions under which Na+ channels can boost conduction of small graded potentials.

It has recently become apparent that in the dendrites or short axons of some neurons, voltage-dependent sodium channels are used not to generate action potentials but to modulate graded potentials; graded potentials carry far more information than do action potentials. A model axon (or dendrite) is described in which sodium channels with kinetics described by equations of the Hodgkin-Huxley type boost conduction of small voltage signals. For a sodium channel density beyond a certain minimum there exists an optimal potential, depolarized with respect to the resting potential, at which there is no steady-state decrement along the axon. For an axon not longer than about 0.7 length constants, small, steady-state deviations from this optimal potential imposed at one end of the axon appear amplified in a graded and stable way at the other end. A small pulse of potential is propagated with amplification and more rapidly than in an axon with a passive membrane. Compared to passive propagation, there will be an improvement in signal-to-noise ratio at the synapse; the axon also acts as a selective frequency filter. The same axon is capable of conducting an action potential.

Animals↗
PubMed1994

Axon contact is associated with modified expression of functional potassium channels in mouse Schwann cells.

In organotypic cultures of mouse dorsal root ganglia, Schwann cells were classed as isolated, that is to say without contact with neurites, or as attached to neurites. It was known that isolated Schwann cells in these cultures display two types of voltage-dependent K+ currents, a fast transient current and a delayed sustained current. In this study, we have investigated outward K+ currents on Schwann cells attached to neurites. These all had a sustained current whose amplitude, timecourse, outward rectification, cumulative inactivation and sensitivity to tetraethylammonium were similar to those of the sustained current on isolated cells. However, the attached cells differed from the isolated cells in that only 15% had a detectable transient current. We suggest three possible explanations for this result: (i) that only cells with reduced transient K+ current move to contact the axon; (ii) that functional expression of these channels is down-regulated by close association with the axon; or (iii) that the channels are lost by transfer to the axon.

Animals↗
PubMed1993

Cranial computed tomography of elderly patients: an evaluation of its use in acute neurological presentations.

We assessed the use of cranial computed tomography (CT) in elderly patients with acute neurological deficit and its influence on patient management. Clinical notes from 100 consecutive CT referrals from geriatric admissions presenting with acute neurological deficit were reviewed and categorized according to clinical presentation. CT results and subsequent therapy were recorded. Twenty of the patients had treatable lesions (in 6 out of 14 patients with signs atypical of stroke and 7 out of 19 patients with acute confusion). These two groups contained 68% of all treatable lesions found. Forty-four scans yielded no new diagnostic information; these included all scans for transient ischaemic attacks and for progression of stroke. The remaining scans yielded information regarding pathology but did not alter patient management. CT is a valuable first-line investigation in elderly patients presenting with signs atypical of stroke and unexplained confusion but may be less useful in patients with other presentations.

Acute Disease↗