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Biomedical subjects

W J Pearce

Publications and source records attributed to W J Pearce.

At least 55 records · Page 3Linked to original sources
PubMed1995

Noradrenaline-mediated contractions of ovine uterine artery: role of inositol 1,4,5-trisphosphate.

To elucidate the role of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) as a second messenger through which noradrenaline regulates contractions of the uterine artery, we present here studies designed to characterize simultaneously the noradrenaline-mediated contractions and Ins(1,4,5)P3 formation in isolated uterine arteries from near-term pregnant sheep. Noradrenaline stimulated a rapid increase of Ins(1,4,5)P3 formation with the peak at 30 second. Simultaneous measurement of noradrenaline-induced contractile responses and Ins(1,4,5)P3 formation revealed a significant linear correlation between these two events. In accordance with the contractile results, the noradrenaline-mediated inositol phosphate accumulation was blocked by prazosin (0.1 microM), but not by yohimbine (0.1 microM). Pre-treatment of tissues with pertussis toxin (200 ng/ml, 3 h) failed to block noradrenaline-induced inositol phosphate accumulation. We conclude that, in the uterine artery of late pregnancy, the alpha 1-adrenoceptor-elicited contraction, at least the initial phasic component, is predominantly mediated by the formation of Ins(1,4,5)P3, leading to release of Ca2+ from intracellular stores.

Animals↗
PubMed1995

Mechanisms of hypoxic cerebral vasodilatation.

Hypoxia activates multiple mechanisms that influence cerebrovascular tone. Through actions on non-vascular cerebral elements, hypoxia stimulates the production of a wide variety of vasodilator metabolites, the most important of which are potassium and hydrogen ions, prostaglandins and adenosine. Hypoxia also promotes the neuronal release of excitatory amino acids, which stimulates overall cerebral metabolism and further enhances the release of vasodilator metabolites. Altogether, the combined action of these metabolites, many of which remain unidentified, account for approximately half the vasodilatation associated with moderate to severe hypoxia. The remaining vasodilatation is attributable to direct effects of hypoxia on cerebral arteries. One component of the direct vascular effects of hypoxia involves the endothelium, which can release at least three different vasodilating factors (prostacyclin, nitric oxide and hyperpolarizing factor) and two different contracting factors (indomethacin-sensitive and indomethacin-resistant) in response to hypoxia. In cerebral arteries, the net contribution of endothelial factors to hypoxic vasodilatation appears to be modest, although the exact profile of factors released by hypoxia appears to depend on both species and artery type. Within the vascular smooth muscle cells of cerebral arteries, hypoxia activates membrane ATP-sensitive potassium channels, resulting in hyperpolarization of the smooth muscle membrane and reduced calcium permeability. In addition, hypoxia also appears capable of retarding flux through the inositol phosphate cascade and reducing the second messenger stimulus for release of intracellular calcium. Both of these latter influences, which may be caused by hypoxic changes in intracellular ATP, ADP and hydrogen ion concentrations, act to lower the free cytosolic calcium concentration available to support contraction. Hypoxia also appears to reduce the calcium sensitivity of contractile proteins. Combined, these mechanisms exert a powerful and multifaceted inhibitory influence on cerebrovascular tone.

Animals↗
PubMed1995

A new model of neonatal stroke: reversible middle cerebral artery occlusion in the rat pup.

Neonatal stroke remains a complex pathophysiologic process that is poorly understood and difficult to investigate. The primary animal model used to study this phenomenon is that of unilateral carotid artery ligation with 2-3 hours exposure to severe hypoxia. A new model of neonatal stroke was developed based on transient middle cerebral artery occlusion without craniectomy. In this model a #6-0 (0.07 mm) nylon filament is passed via the carotid artery to occlude reversibly the middle cerebral artery for 4 hours under conditions of normoxia in 14- to 18-day-old spontaneously hypertensive rat pups. After removal of the filament and reperfusion for 24 hours, the infarct volume was determined using the mitochondrial stain, 2,3,5-triphenyltetrazolium chloride. Using this technique, a neocortical and caudoputamenal infarct affecting 49% of hemispheric volume that measured 180 +/- 29 mm3 (hemisphere volume = 359 +/- 16 mm3, mean +/- SEM) was created in 90% of animals (n = 8) undergoing this procedure. This model has the advantage of being relatively noninvasive, of not requiring global exposure of brain to hypoxia, and of using temporary rather than permanent occlusion. This technique should improve the ability to study the acute and long-term pathophysiology of neonatal stroke, particularly the phenomenon of reperfusion injury, as well as its sequelae in the developing nervous system.

Animals↗
PubMed1995

Effects of maturation on cell water, protein, and DNA content in ovine cerebral arteries.

The present study examined the effects of maturation on base-soluble protein, DNA content, and intracellular water volume in ovine cerebral arteries to evaluate these variables as references for normalization of levels of cellular constituents in studies of vascular maturation. With maturation, base-soluble protein per unit wet weight (measured by using the Bradford method) increased by 27% to 46%, and cell volume (estimated as the ratio of cell water to DNA) increased by 53% to 97%. However, intracellular water per unit wet weight (calculated as the difference between total water measured by dehydration and extracellular water measured by using the extracellular marker [57Co]EDTA) increased by only 1% to 16%, because of maturation-related hypertrophy (quantitated histologically) combined with decreased cell number per unit wet weight (DNA content, quantitated by using Hoechst dye, decreased by 32% to 41%). In addition, the effects of maturation were artery specific: maturational increases in cell water and volume were more pronounced in common carotid than in cerebral arteries. These findings demonstrate that different methods of normalization can produce quantitatively opposite results in maturation-related studies. For cellular constituents, the most physiologically relevant normalization is relative to intracellular water volume, which yields units of apparent intracellular concentration. Because the relative content of intracellular water per unit wet weight changes little with maturation, normalization of levels of cellular constituents relative to wet weight provides a reasonable index of intracellular concentration. Clearly, no single approach is optimal for all studies, and the method of normalization should be carefully considered with regard to effects of maturation on vascular composition.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗
PubMed1995

Maturation modulates serotonin- and potassium-induced calcium-45 uptake in ovine carotid and cerebral arteries.

Neonatal vulnerability to intracranial hemorrhage is often attributed to a relative inability of immature cerebral arteries to contract. Because this depressed contractility may involve age-related differences in cerebrovascular calcium handling, the present study examined age-related differences in cerebral artery contractility and its dependence on extracellular calcium from 24 newborn lambs and 36 adult sheep. Contractile tensions and 45Ca uptakes were measured under baseline conditions and as a function of time during stimulation by both receptor-dependent (100 microM serotonin) and receptor-independent (122 mM K+) mechanisms of contraction in endothelium denuded newborn (N) and adult (A) ovine middle cerebral (MCA) and common carotid (COM) arteries. Maximum contractile responses to potassium averaged 4.5 +/- 0.2 (N-COM), 5.8 +/- 0.9 (A-COM), 3.0 +/- 1.1 (N-MCA), and 3.1 +/- 0.6 (A-MCA) g. Corresponding averages for responses to serotonin were 7.2 +/- 0.8, 7.3 +/- 1.1, 3.6 +/- 0.1, and 3.6 +/- 0.2; except for COM responses to potassium, contractile responses were little affected by maturation in either artery type. At baseline, uptakes averaged 0.39 +/- 0.04 (N-MCA), 0.33 +/- 0.04 (A-MCA), 0.25 +/- 0.03 (N-COM), and 0.14 +/- 0.01 (A-COM) mumol Ca/g dry weight/min. Maximum increases in calcium uptake produced by potassium depolarization averaged 231 +/- 19% (N-MCA), 152 +/- 13% (A-MCA), 156 +/- 11% (N-COM), and 140 +/- 14% (A-COM) above baseline. Corresponding increases produced by 100 microM serotonin averaged 201 +/- 15% (N-MCA), 129 +/- 23% (A-MCA), 143 +/- 20% (N-COM), and 145 +/- 18% (A-COM).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

L-NAME reduces infarct volume in a filament model of transient middle cerebral artery occlusion in the rat pup.

The importance of nitric oxide (NO) during focal cerebral ischemia remains controversial as studies have suggested both a neurotoxic and neuroprotective role. In the 7 d old rat pup, NG-nitro-L-arginine, a nitric oxide synthase inhibitor, reduced infarct volume in a model of unilateral carotid ligation with 2.5 h exposure to 8% O2. The current study examined whether NO is neurotoxic in a filament model of transient middle cerebral artery occlusion (MCAO) in the 14-18-d-old rat pup. We developed a reproducible filament model of transient MCAO in 14-18-d-old spontaneously hypertensive rats (35 g) by passing a no. 6-0 (0.07-mm) nylon filament via the carotid artery to occlude the middle cerebral artery for 4 h under normoxic conditions. After filament removal and reperfusion for 24 h, we determined infarct volume using the mitochondrial stain 2,3,5-triphenyltetrazolium chloride. NO synthesis was inhibited using NG-nitro-L-arginine methyl ester (L-NAME) at a dose of 3 mg/kg, intraperitoneally, 1 h before MCAO. We measured infarct volume in control (n = 7) and L-NAME (n = 7) groups. L-NAME reduced infarct volume by 55% (p < 0.01). In the control group, infarct volume (180 +/- 29 mm3) averaged 49 +/- 7% of the left hemisphere (359 +/- 16 mm3). In the L-NAME-treated group, infarct volume (77 +/- 19 mm3) was 22 +/- 5% of the left hemispheric volume (344 +/- 2 mm3). These findings support earlier studies that used models of neonatal hypoxic-ischemic brain injury and suggest a neurotoxic role of NO. They extend these observations by demonstrating a significant reduction in infarct volume in a stroke model in the immature rat pup.

Animals↗
PubMed1994

Endothelium-derived relaxing factor and cyclic GMP-dependent vasorelaxation in human chorionic plate arteries.

Endothelium derived relaxing factor (EDRF), now widely believed to be nitric oxide (NO), may play an important part in the control of fetoplacental vascular tone. To further explore this role we have determined the relaxation responses to exogenous NO and examined the temporal relationship between intracellular concentrations of cyclic GMP and vascular tone in isolated ring segments of human chorionic plate arteries. We have also determined the dose relations for the contractile agonists serotonin and the thromboxane analog U46619. Lastly, we have explored the relaxation responses to a wide range of agents known to elicit EDRF release in other vascular beds. Chorionic plate arteries relaxed significantly to exogenous NO with concomitant increases in cyclic guanosine monophosphate over basal values. ED50s for serotonin and U46619 were 1.48 x 10(-6) M and 3.39 x 10(-8) M respectively. The ED50 for NO derived from S-nitroso-N-acetyl-penicillamine was 1.28 x 10(-6) M. Endothelium-intact segments of chorionic plate arteries pre-contracted with either serotonin or U46619 failed to relax significantly to acetylcholine, adenosine diphosphate, A23187, bradykinin, and histamine and only minimally to substance P. We suggest that EDRF is likely to be important in the control of placental vascular tone, but that it is not possible to demonstrate its action in an unperfused experimental system.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1994

Dual effects of L-NAME during transient focal cerebral ischemia in spontaneously hypertensive rats.

The role of nitric oxide (NO) in ischemic neuronal injury is unclear. In permanent focal ischemia models, NO release has been reported to be both neuroprotective, by virtue of actions to improve cerebral blood flow (CBF) within ischemic tissue, and neurotoxic. Very little attention has been given to determining the role of NO in transient focal ischemia. In the present studies, low-dose NO inhibition using NG-nitro-L-arginine methyl ester (L-NAME; 0.1 mg/kg bolus, 0.01 mg.kg-1.min-1 iv) reduced infarct volume after 180 min of middle cerebral arterial occlusion (MCAO) and 120 min of reperfusion as measured via 2,3,5-triphenyltetrazolium chloride by 55% (P < 0.0001). Similar reductions occurred whether L-NAME was given throughout MCAO-reperfusion or just 30 or 60 min before reperfusion. L-NAME reduced CBF in the area of infarction at 30 and 180 min of MCAO by 36 and 33% (P < 0.02). In contrast, 15 min into reperfusion, L-NAME increased CBF in the area of infarction by 69% (P < 0.03) and by 27% in the contralateral homologous right hemisphere. Although vascular effects are present, these findings suggest a neurotoxic role for NO primarily during reperfusion after transient focal ischemic injury.

Animals↗
PubMed1994

Effects of maturation on alpha-adrenergic receptor affinity and occupancy in small cerebral arteries.

The present experiments examine the hypothesis that changes in receptor affinity and occupation mediate maturational changes in norepinephrine sensitivity in small cerebral arteries. In second-order (2B) and fourth-order (4B) branch middle cerebral artery segments from newborn and adult sheep, we first found that a stretch ratio based on artery diameter better estimated optimal prestretch than did passive tension. Next, we determined norepinephrine dose-response relations before and after prazosin, yohimbine, and benextramine. Prazosin competitively blocked contractions to norepinephrine, but yohimbine had no effect, indicating that alpha 1-adrenoceptors mediated contraction. Norepinephrine sensitivity [determined from the -log of the half-maximal effective dose (pD2)], maximal response, and binding affinity all decreased with age in 4B but not 2B segments. Receptor occupancy at the pD2 increased with age only in 2B segments. In conclusion, maturation of ovine middle cerebral arteries involves branch-specific changes in affinity and receptor occupation of the alpha 1-adrenoceptors that mediate contractile responses to norepinephrine. Age-related changes in receptor density and/or intrinsic efficacy probably are involved also.

Adrenergic alpha-Antagonists↗
PubMed1994

Pregnancy alters cerebrovascular adaptation to high-altitude hypoxia.

We have previously shown alterations in cerebrovascular composition, contractility, and endothelial function in normoxic pregnant (P) and chronically hypoxic nonpregnant (HNP) adult sheep compared with nonpregnant normoxic controls (NP). This study focuses on a fourth group, pregnant sheep exposed to chronic high-altitude hypoxia (HP) (110 days at 3,820 m). The combined challenges of pregnancy and high-altitude hypoxia resulted in significant alterations in cerebrovascular function that were not simply the summation of the responses seen in the P and HNP animals. Compared with NP, HP arteries had increased protein content and increased intracranial arterial wall thickness. Both P and HP arteries showed increased contractility, while HNP artery maximum tensions were depressed. Endothelial function was depressed in the P common carotid and was enhanced in all HNP arteries. In contrast, an increased endothelial response was shown only in the HP common carotid. Thus, for contractility, the effects of pregnancy predominated in the HP arteries. For endothelial function, hypoxia effects were dominant in the common carotid but not in the intracranial arteries. Clearly, cerebrovascular characteristics are dynamic in nature, with artery-specific responses to both pregnancy and hypoxia.

Adaptation, Physiological↗
PubMed1994

Maturation enhances the sensitivity of ovine cerebral arteries to the ATP-sensitive potassium channel activator lemakalim.

A wide variety of previous studies have demonstrated that arterial reactivity and contractility change dramatically during maturation. In light of recent findings that binding sites for glibenclamide, a ligand for ATP-sensitive potassium (KATP) channels, become more abundant with age in many tissues, the present studies examine the hypothesis that maturational changes in vascular reactivity involve changes in arterial electrophysiologic characteristics. To test this hypothesis, we determined the dose-response relation to lemakalim, a selective activator of KATP channels, in isolated endothelium-denuded segments of the second (2B, internal diameter approximately 200 microns) and fourth (4B, internal diameter approximately 125 microns) branches of middle cerebral arteries taken from newborn (3-7 d old) and adult sheep. At 100 microM, lemakalim completely relaxed serotonin-induced tone in all arteries. However, -log ED50 values were 29 to 43 times greater in adult (2B, 7.15 +/- 0.38; 4B, 6.61 +/- 0.42) than in newborn (2B, 5.52 +/- 0.25; 4B, 5.15 +/- 0.24) segments. Correspondingly, Hill values were significantly smaller in adults (2B, 0.47 +/- 0.17; 4B, 0.71 +/- 0.30) than in newborns (2B, 1.40 +/- 0.35; 4B, 3.30 +/- 0.92). These findings demonstrate that KATP channels are less sensitive to activation in newborn than in adult cerebral arteries. Given the important influence of KATP channels on vascular tone, and their possible role in many cardiovascular responses, the present data suggest that maturational increases in the activity of KATP channels contribute significantly to age-related changes in cerebrovascular contractility.

Adenosine Triphosphate↗
PubMed1994

Effects of maturation on cyclic GMP-dependent vasodilation in ovine basilar and carotid arteries.

The present experiments examine the effects of maturation on cyclic GMP (cGMP)-mediated vasodilation in 688 segments of common carotid (COM) and basilar (BAS) arteries taken from newborn (3- to 7-d-old) and nonpregnant adult sheep. The main finding is that maximum efficacy for relaxation decreased with maturation in both artery types for the nitric oxide releasing vasodilators S-nitroso-N-acetyl-penicillamine and nitroglycerin. These decreases could not be explained by changes in the -log ED50 concentrations for either vasodilator. Determination of the time course of cGMP responses to S-nitroso-N-acetyl-penicillamine or nitroglycerin at 10 microM revealed that the peak cGMP responses to these agents (range: 5.3 +/- 0.8 to 8.3 +/- 1.6 pmol/mg of protein) also did not vary significantly with age. However, cGMP attained peak values more rapidly in adult (COM: 50 s; BAS 30 s) than in newborn (COM: 60-80 s: BAS, 40-60 s) segments and returned to baseline more slowly in newborn than in adult segments, suggesting that maturation accelerates cGMP turnover. Correspondingly, baseline levels of cGMP were higher in newborn (COM: 1.0 +/- 0.1; BAS: 3.3 +/- 0.5 pmol/mg of protein) than in adult (COM: 0.3 +/- 0.1; BAS: 1.7 +/- 0.2 pmol/mg of protein) segments. Despite these differences in cGMP time course, rates of relaxation in response to S-nitroso-N-acetyl-penicillamine and nitroglycerin did not vary significantly with age, indicating that the temporal relation between cGMP and relaxation is different in newborn and adult arteries.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Low dose L-NAME reduces infarct volume in the rat MCAO/reperfusion model.

In a variety of recent studies, inhibitors of nitric oxide (NO) synthesis have ameliorated neuronal injury during permanent focal cerebral ischemia, suggesting that NO may contribute to ischemic damage. In other studies, however, these inhibitors increased infarct volume during permanent middle cerebral artery occlusion (MCAO). One complication in these studies was that high-dose NO synthase inhibitors increased mean arterial blood pressure (MAP) by 20-30 mm Hg. Thus, it is possible that variations in the effects of NO synthesis inhibitors on infarct volume could be related to effects of these inhibitors on MAP and cerebral perfusion during or after ischemia. The present study compared the effects of control (Ringer's lactate solution) versus low-dose NO inhibition (0.1 mg/kg bolus followed by 0.01 mg/kg/min) on cerebral infarct volume using L-NAME (NG-nitro-L-arginine methyl ester) administered during a 1-h baseline period, 3-h of MCAO, and 2 h of reperfusion in the spontaneously hypertensive rat. Infarct volume was determined using the TTC (2,3,5-triphenyltetrazolium chloride) method performed 5 h after onset of occlusion. L-NAME reduced infarct volume by 55%. In the control group (n = 7), infarct volume measured 116 +/- 4 (SEM) mm3 which was 29 +/- 1% of the left hemispheric volume (400.5 +/- 0.3 mm3). In the L-NAME group (n = 7), infarct volume measured 53 +/- 8 mm3 which was only 13 +/- 2% of the left hemispheric volume (400.4 +/- 0.5 mm3).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Cerebrovascular adaptations to high-altitude hypoxemia in fetal and adult sheep.

In the fetus and infant, high-altitude hypoxemia is associated with increased cerebrovascular morbidity. To test the hypothesis that this increased morbidity involves changes in cerebrovascular endothelial and smooth muscle function, we examined middle cerebral, posterior communicating, basilar, and common carotid arteries obtained from 23 normoxic fetuses, 19 hypoxemic fetuses maintained at high altitude (3,820 m) from 30 days gestation to near term (approximately 143 days), 55 normoxic non-pregnant adults, and 24 hypoxemic nonpregnant adults maintained at the same altitude and duration as the hypoxemic fetuses. Long-term hypoxemia was associated with several significant changes in both fetal and adult arteries, including a generalized increase in base-soluble protein (5-50%), a depression of the maximum potassium-induced tensions (16-49%), and a depression of the relaxation responses to S-nitroso-N-acetylpenicillamine (1-11%), which releases nitric oxide into solution upon hydration. Altitude acclimatization significantly enhanced amine-to-potassium ratios (the ratio of tension produced by 10 microM serotonin with 20 microM histamine to that produced by 122 mM potassium) only in adult cerebral arteries (51-87%) and significantly depressed potassium-induced stresses (up to 41%) and serotonin/histamine-induced tensions (20-37%) only in fetuses. Endothelium-dependent relaxations to A23187 were significantly depressed in hypoxemic fetuses (4-11%) but were significantly enhanced in hypoxemic adults (2-14%). We conclude that chronic hypoxemia depresses both vascular smooth muscle and endothelial function to a greater extent in fetal than in adult cerebral arteries and that this effect could contribute to the greater postnatal vulnerability to asphyxic and hypertensive insults seen in hypoxemic neonates.

Acclimatization↗
PubMed1992

Differential cerebrovascular and metabolic responses in specific neural systems elicited from the centromedian-parafascicular complex.

The effect of electrical stimulation of the centromedian-parafascicular complex on local cerebral blood flow and local cerebral glucose utilization was investigated in anesthetized, paralysed and ventilated rats. Local cerebral blood flow and local cerebral glucose utilization were measured in separate groups of animals using the autoradiographic (14C)iodoantipyrine and (14C)2-deoxyglucose methods, respectively. Because of the well-established centromedian-parafascicular complex neuroanatomical connections, three functional neuronal systems were analysed and compared: the extrapyramidal motor system the limbic system and the reticular formation, also known as the ascending activating system. Cortical regions not included in the limbic system were considered separately. The validity of comparisons between changes in local cerebral blood flow and local cerebral glucose utilization across the brain was verified by assessing the reactivity and stability of the cortical blood flow during long-term centromedian-parafascicular complex stimulation. Centromedian-parafascicular complex stimulation elicited a marked but heterogeneous increase in local cerebral blood flow in 50 of the 52 cerebral structures measured. The most pronounced increases were seen in the lateral habenular nucleus (331 +/- 30% of control), the zona incerta (400 +/- 55%), the mesencephalic reticular formation (415 +/- 122%) and the parietal cortex (211 +/- 35%). In contrast, local cerebral glucose utilization remained statistically unchanged (P greater than 0.05) in 28 of these 50 individual brain regions during centromedian-parafascicular complex stimulation. The most pronounced increases in local cerebral glucose utilization were seen in the zona incerta (123 +/- 28%) and the mesencephalic reticular formation (193 +/- 26%). Local cerebral blood flow and local cerebral glucose utilization were linearly related in unstimulated controls, considering either all brain regions taken as a whole or the three systems separately. The significant increase in the slopes of the regression line between local cerebral blood flow and local cerebral glucose utilization for the reticular formation and the limbic system during centromedian-parafascicular complex stimulation indicates, however, that the coupling mechanisms for these systems, but not for the extrapyramidal motor system, were reset. The local cerebral blood flow to local cerebral glucose utilization ratio was heterogeneous in controls and differentially increased during centromedian-parafascicular complex stimulation, being markedly pronounced in the parietal cortex and in the reticular formation. We conclude that these results, for the first time, provide evidence that, the functionally well-defined neural networks may have different mechanisms whereby changes in vascular and metabolic demands are regulated.

Anesthesia↗