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W J Pearce

Publications and source records attributed to W J Pearce.

At least 37 records · Page 2Linked to original sources

High altitude, hypoxic-induced modulation of noradrenergic-mediated responses in fetal and adult cerebral arteries.

In response to high altitude long-term hypoxemia, the cerebral arteries of adult and fetal sheep show decreased contractile responses to norepinephrine and other agonists. In this review, we examine some of the presynaptic and postsynaptic contractile mechanisms that might account for these changes. In addition, we examine cerebral vessel relaxation and the role of pregnancy in altering these responses. In general, high altitude hypoxia is associated with augmented or "upregulation" of presynaptic functions. In contrast, postsynaptic functions tend to be significantly depressed or "downregulated." The results emphasize the role of high altitude, long-term hypoxemia in modulating adrenergic-mediated signal transduction in the cerebral vasculature. They specifically highlight the profound differences in acclimatization responses between common carotid and intracranial arteries, as well as the significant differences between responses in the fetus and adult.

Acclimatization↗

Regulation of Ca2+ sensitization by PKC and rho proteins in ovine cerebral arteries: effects of artery size and age.

G protein-regulated Ca2+ sensitivity of vascular contractile proteins plays an important role in cerebrovascular reactivity. The present study examines the intracellular mechanisms that govern G protein-regulated Ca2+ sensitivity in cerebral arteries of different size and age. We studied beta-escin-permeabilized segments of common carotid, basilar, and middle cerebral arteries from nonpregnant adult and near-term fetal sheep. Activation of protein kinase C (PKC) by (-)-indolactam V or a phorbol ester produced receptor-independent increases in Ca2+ sensitivity. Such increases were more marked in immature arteries and were inversely correlated with artery size in both mature and immature arteries. However, inhibitors of PKC did not significantly affect increases in Ca2+ sensitivity in responses to either serotonin (5-hydroxytryptamine, 5-HT) or guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS). Alternatively, deactivation of rho p21, a small G protein associated with Rho kinase, by exotoxin C3 fully prevented increases in Ca2+ sensitivity in responses to 5-HT or GTPgammaS in both adult and fetal arteries of all types. Neither inhibitors of PKC nor exotoxin C3 altered baseline Ca2+ sensitivity. We conclude that patterns of receptor- and/or G protein-mediated modulation of Ca2+ sensitivity are dependent on an intracellular pathway that involves activation of small G proteins and Rho kinase. In contrast, PKC has little, if any, role in agonist-induced Ca2+ sensitization under the present experimental conditions.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Effects of maturation, artery size, and chronic hypoxia on 5-HT receptor type in ovine cranial arteries.

To test the hypothesis that variations in cerebrovascular reactivity to 5-HT among arteries of different size or type, during maturation, or during acclimatization to high altitude involve differences in serotonergic receptor subtype, we determined relative agonist potency orders and antagonist affinities in common carotid (Com), main branch middle cerebral (Main), and second branch middle cerebral (2BR) arteries from term fetal lambs and nonpregnant adult sheep acclimatized at sea level or at an altitude of 3,820 m for approximately 110 days. In normoxic adult Com segments, agonist potency order was 5-hydroxytryptamine (5-HT) > 5-carboxamidotryptamine (5-CT) >/= 8-hydroxy-2(di-n-propylamino)tetraline (8-OH-DPAT); sumatriptan (Suma) produced no contractile response; and antagonist dissociation constant (pKb) values were 9.4 and 9.5 for ketanserin against 5-HT and 5-CT, 7.5 for GR-127935 against 5-HT, and 7.2 for SB-206553 against 5-HT. In normoxic adult Main segments, agonist potency order was 5-HT > 5-CT >/= Suma >/= DPAT, and pKb values were 9.1 and 9.2 for ketanserin against 5-HT and 5-CT and 7.4 and 8.5 for GR-127935 against 5-HT and Suma, respectively. In the 2BR segments from normoxic adults, agonist potency order was 5-CT > 5-HT > Suma > DPAT and pKb values were 7.4 and 7.2 for ketanserin against 5-HT and 5-CT and 10.0 and 8.7 for GR-127935 against 5-HT and Suma, respectively. Compared with normoxic adults, none of these values were significantly different in hypoxic adults and in fetuses only the pKb values for ketanserin against 5-HT in the 2BR segments (8.8) were greater. From these results we propose that the ratio of 5-HT2 to 5-HT1 receptors is greatest in the Com and decreases progressively to its smallest values in 2BR or smaller segments. Because this gradient appears stable and relatively resistant to the effects of maturation and chronic hypoxia, changes in reactivity associated with these perturbations may involve alterations in receptor density and/or coupling efficiency for 5-HT in ovine cranial arteries.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Developmental changes in the calcium sensitivity of rabbit cranial arteries.

The present experiments examine developmental changes in cerebrovascular Ca2+ sensitivity. Common carotid (COM), basilar (BAS) and femoral (FA) arteries from adult (n = 16), 8- to 9-day-old (n = 15) and 24- to 25-day-old rabbits (n = 12) were denuded of the endothelium and permeabilized with beta-escin. Bath calcium concentrations were controlled via EGTA-Ca2+ buffer solutions. Adult pCa-force relations were right-shifted relative to those of 8- to 9-day-old rabbits but were similar to those of 24- to 25-day-olds. Adult pD2 (-log ED50) values for Ca2+ averaged 6.36 +/- 0.03 (COM), 6.77 +/- 0.04 (BAS) and 6.40 +/- 0.04 (FA). Corresponding 8- to 9-day-old values were 6.85 +/- 0.03, 7.08 +/- 0.08 and 6.76 +/- 0.05. In all arteries studied, the addition of 5-hydroxytryptamine (5-HT) subsequent to contraction by a constant submaximal (EC30) concentration of Ca2+ produced a dose-dependent and GDP3S-sensitive increase in tension attributable to an increase in Ca2+ sensitivity. The magnitudes of 5-HT-induced increases in Ca2+ sensitivity were significantly greatest in 8- to 9-day-old rabbits, intermediate in 24- to 25-day-old rabbits, and least in adults. GTPgammaS mimicked the effects of 5-HT and prevented further increases in Ca2+ sensitivity induced by 5-HT in all arteries from all age groups. GDPbetaS completely reversed all effects of 5-HT on Ca2+ sensitivity. From these data we conclude that baseline Ca2+ sensitivity is elevated in newborn relative to adult rabbits, at least in femoral, common carotid and basilar arteries. In these arteries, 5-HT can increase Ca2+ sensitivity via a G-protein-dependent mechanism which is more effective in neonatal than adult arteries. These effects of maturation on vascular Ca2+ sensitivity may play an important role in developmental changes in vascular reactivity.

Aging↗

Maturational modification of hypoxic relaxation in ovine carotid and cerebral arteries: role of endothelium.

In light of evidence that hypoxia inhibits cerebrovascular calcium influx and that the contractile importance of calcium influx is greater in neonatal than adult cerebral arteries, the present studies evaluate the hypothesis that the mechanisms involved in hypoxic cerebral vasodilation are different in newborn and adult cerebral arteries. Intact segments of ovine common carotid (COM) and middle cerebral (MCA) arteries from 3- to 7-day-old newborn lambs and nonpregnant adults were mounted in tissue baths, contracted with 1 microM 5-HT, and exposed to hypoxia for 20 min (PO2 approximately 12 Torr). In endothelium-intact arteries, the magnitude of relaxation was similar and near maximal in all groups (range 82-92%) and did not vary with either age or artery type. However, the magnitude of hypoxic relaxation after endothelium removal was attenuated more in adult (COM 37 +/- 6%, MCA 53 +/- 7%) than in newborn (COM 75 +/- 5%, MCA 93 +/- 1%) arteries. In addition, endothelium-dependent responses to A23187, which like hypoxia increases endothelial cell concentrations of calcium, were attenuated in immature compared to mature arteries. Reductions of extracellular calcium concentration from 1.6 to 0.8 mM eliminated all age-dependent differences in relaxation responses to hypoxia. Together these data demonstrate that in newborn arteries, the endothelium-dependent component of hypoxic vasodilation is minimal, especially in intracerebral arteries, and robust relaxant responses to hypoxia depend on the high sensitivity of immature smooth muscle cells to decreases in PO2. In contrast, in adult arteries the smooth muscle sensitivity to decreases in PO2 is minimal and the endothelial component of hypoxic relaxation is particularly important in responses to hypoxia.

Adenosine Diphosphate↗

Maturation alters the contractile role of calcium in ovine basilar arteries.

The present studies examine how agonist-induced increases in cytosolic Ca2+ concentration and sensitivity vary with maturation. Basilar arteries from term fetal (138-141 d) and nonpregnant adult sheep were denuded of endothelium, mounted for measurements of contractile tension, and then loaded with Fura-2 to enable estimation of cytosolic Ca2+ responses to both potassium and serotonin (5-hydroxytryptamine, 5-HT). In response to potassium, normalized values of intracellular Ca2+ and tension increased in parallel in both fetal and adult preparations; no age-related differences were apparent. In contrast, 5-HT increased Ca2+ sensitivity significantly more in fetal than in adult arteries. In the absence of extracellular Ca2+, 5-HT increased cytosolic Ca2+ in adult but not fetal arteries. In addition, responses to repeated applications of 5-HT in the absence of extracellular Ca2+ were exhausted more rapidly in fetal than in adult arteries. We interpret these data to indicate that vascular maturation involves important shifts in the mechanisms mediating cerebrovascular pharmacomechanical coupling. Specifically, the data suggest that normal development involves a reduction in the Ca2+ sensitizing effects of agonists with parallel increases in the agonist-induced intracellular Ca2+ release. In so doing, these studies offer one possible reason why vascular reactivity varies dramatically with age. From a pathophysiologic perspective, these studies also advance the possibility that failure to shift from the increased Ca2+ sensitivity typical of immature arteries may lead to vascular hyperreactivity in adult arteries.

Animals↗

Core and penumbral nitric oxide synthase activity during cerebral ischemia and reperfusion.

BACKGROUND AND PURPOSE: The present studies examined the hypothesis that the distribution of cerebral injury after a focal ischemic insult is associated with the regional distribution of nitric oxide synthase (NOS) activity. METHODS: Based on previous studies that certain anatomically well-defined areas are prone to become either core or penumbra after middle cerebral artery occlusion (MCAO), we measured NOS activity in these areas from the right and left hemispheres in a spontaneously hypertensive rat filament model. Four groups were studied: (1) controls (immediate decapitation); (2) 1.5 hours of MCAO with no reperfusion (R0); (3) 1.5 hours of MCAO with 0.5 hour of reperfusion (R0.5); and (4) 1.5 hours of MCAO with 24 hours of reperfusion (R24). Three groups of corresponding isoflurane sham controls were also included: 1.5 (S1.5) or 2 (S2.0) hours of anesthesia and 1.5 hours of anesthesia+24 hours of observation (S24). RESULTS: Control core NOS activity for combined right and left hemispheres was 129% greater than penumbral NOS activity (P<0.05). Combined core NOS activity was also greater (P<0.05) in the three sham groups: 208%, 122%, and 161%, respectively. In the three MCAO groups, ischemic and nonischemic core NOS remained higher than penumbral regions (P<0.05). However, NOS activity was lower in the ischemic than in the nonischemic core in all three groups: R0 (29% lower), R0.5 (48%), and R24 (86%) (P<0.05). Addition of cofactors (10 micromol/L tetrahydrobiopterin, 3 micromol/L flavin adenine dinucleotide, and 3 micromol/L flavin mononucleotide) increased NOS activity in all groups and lessened the decrease in ischemic core and penumbral NOS. CONCLUSIONS: Greater NOS activity in core regions could explain in part the increased vulnerability of that region to ischemia and could theoretically contribute to the progression of the infarct over time. The data also suggest that NOS activity during ischemia and reperfusion could be influenced by the availability of cofactors.

Anesthetics, Inhalation↗

Activator calcium and myocardial contractility in fetal sheep exposed to long-term high-altitude hypoxia.

We studied myocardial contractility in fetal sheep from ewes exposed to approximately 112 days of hypoxia at high altitude (3,820 m). We measured the inotropic response to extracellular Ca2+ concentration ([Ca2+]o, 0.2-10 mM) and ryanodine (10(-10) to 10(-4) M) in isometrically contracting papillary muscles and quantified dihydropyridine (DHPR) and ryanodine (RyR) receptors. In hypoxic fetuses, curves describing the force-[Ca2+]o relationship were shifted left, and the top plateaus were decreased by approximately 35% in both left and right ventricles. In normoxic and hypoxic fetuses, ryanodine (10(-4) M) reduced maximum active tension (Tmax) to approximately 25-40% of baseline values, indicating that the sarcoplasmic reticulum was the chief source of activator Ca2+ and that Ca2+ influx alone was not sufficient to activate a contraction of normal amplitude. Hypoxia resulted in a lower Tmax in the right ventricle and a lower maximum rate of rise in the left ventricle after treatment with ryanodine. DHPR number did not change, but RyR number and the RyR/DHPR in both ventricles were higher in hypoxic fetuses. We conclude that hypoxia decreases contractility, possibly by reducing the availability of activator Ca2+. Further studies are needed to directly measure the Ca2+ current and intracellular Ca2+ transient and to examine myofilament protein and adenosinetriphosphatase activity.

Altitude↗

Physiological variations in ovine cerebrovascular calcium sensitivity.

Cerebrovascular reactivity to biogenic amines varies in relation to both arterial diameter and age. The present study examines the hypothesis that these patterns of reactivity are secondary to corresponding variations in the Ca2+ sensitivity of the contractile proteins. To test this hypothesis, we permeabilized segments of common carotid (Com), basilar, main branch middle cerebral, and second-branch middle cerebral (MCA-B) arteries from nonpregnant adult and near-term fetal sheep using beta-escin. Permeabilization methods were carefully validated and adjusted for each artery type. Baseline myofilament Ca2+ sensitivity in both adults and fetuses increased significantly from the Com to the MCA-B and was generally higher in fetuses than in adults. Serotonin dose dependently increased Ca2+ sensitivity via a G protein-dependent mechanism in all arteries. The magnitudes of this effect did not vary among artery types but were significantly greater in fetal than in adult arteries. This effect of serotonin was mimicked by guanosine 5'-O-(3-thiotriphosphate), a nonhydrolyzable analog of guanosine 5'-triphosphate, and its effects were also much greater in fetal than in adult arteries. We conclude that patterns of cerebrovascular reactivity to biogenic amines were determined, at least in part, by underlying variations in baseline myofilament Ca2+ sensitivity and/or its alteration by G protein-dependent mechanisms.

Animals↗

Maturation alters cerebral NOS kinetics in the spontaneously hypertensive rat.

Using 14C-labeled arginine to 14C-labeled citrulline conversion assays in brain homogenates from 14- to 18-day-old and adult spontaneously hypertensive rats, we tested the hypotheses that maturation increases neuronal nitric oxide synthase (nNOS) activity and that this increase involves changes in cofactor availability and/or nNOS kinetics. nNOS activity (in pmol x mg(-1) x min(-1)) was 46% higher in adults (19 +/- 2) than in pups (13 +/- 1). The addition of 264 microM calmodulin (CaM), 3 microM FAD, 3 microM flavin adenine mononucleotide (FMN), and 10 microM tetrahydrobiopterin (BH4) increased NOS activity by 3, 46, 45, and 88% in pups and by 19, 40, 36, and 102% in adults, respectively. All cofactor effects were significant except for CaM in the pup homogenates. Cofactor effects were not significantly different between pup and adult homogenates, except for BH4, which increased absolute NOS activity more in adults than in pups. Values of maximal enzyme velocity (Vmax) for nNOS in the absence of added cofactors were greater in adults than in pups (104 +/- 5 vs. 53 +/- 3, P < 0.05). Addition of 3 microM FAD or 3 microM FMN increased pup Vmax values to 68 +/- 2 and 99 +/- 5, respectively, but had no effect in adults. BH4 did not affect Vmax in either group. Control values of the Michaelis-Menten constant (Km) for L-arginine were greater (P < 0.05) in pups (5.7 +/- 0.4 microM) than in adults (4.3 +/- 0.2 microM) and were significantly reduced by 10 microM BH4 to 3.8 +/- 0.2 and 2.9 +/- 0.1 microM, respectively. Neither FAD nor FMN affected Km values in either group. The results indicate that endogenous nNOS cofactor levels are not saturating in either pups or adults, changes in cofactor levels differentially affect NOS kinetics in pups and adults, and age-related differences in NOS activity result from fundamental differences in NOS kinetics. These findings support the general hypothesis that the increased vulnerability to ischemic stroke associated with maturation is due in part to corresponding increases in the capacity for nitric oxide synthesis.

Aging↗

Cardiac beta-adrenergic receptor function in fetal sheep exposed to long-term high-altitude hypoxemia.

In this study, we hypothesized that a reduction in beta-adrenergic receptor number or a decrease in functional coupling of the receptor to the adenylate cyclase system may be responsible for the blunted inotropic response to isoproterenol observed in fetal sheep exposed to high altitude (3,820 m) from 30 to 138-142 days gestation. We measured the contractile response to increasing doses of isoproterenol and forskolin in papillary muscles from both ventricles, estimated beta-adrenergic receptor density (Bmax) and ligand affinity (Kd) using [125I]iodocyanopindolol, and measured adenosine 3',5'-cyclic monophosphate (cAMP) levels before and after maximally stimulating doses of isoproterenol and forskolin. Left ventricular wet weight was unchanged, but right ventricular weight was 20% lower than controls. At the highest concentration of isoproterenol (10 microM), maximum active tension was 32 and 20% lower than controls in hypoxemic left and right ventricles, respectively. The contractile response to forskolin was severely attenuated in both hypoxemic ventricles. Bmax was unchanged in the left ventricle, but increased by 55% in the hypoxemic right ventricle. Kd was not different from controls in either ventricle. Basal cAMP levels were not different from controls, but isoproterenol-stimulated and forskolin-stimulated cAMP levels were 1.4- to 2-fold higher than controls in both hypoxemic ventricles. The results suggest mechanisms downstream from cAMP in the beta-adrenergic receptor pathway are responsible for the attenuated contractile responses to isoproterenol.

Adrenergic beta-Agonists↗

Intracranial-extracranial differences in the Ca2+ sensitivity of rabbit arteries.

We have previously demonstrated that the ratio of calcium uptake to force production varies widely with age, artery size, and method of contraction in cerebral arteries. The present experiments were conducted to examine the possibility that these differences involve corresponding variations in contractile force-calcium relations. Common carotid (COM), basilar (BAS), and middle cerebral (MCA) arteries from adult were denuded of endothelium and mounted in vitro for measurement of contractility. Following equilibration at optimum resting diameter, the arteries were permeabilized (beta-escin, 50 microg/ml) and depleted of intracellular Ca2+ by treatment with 1 microM A23187. Ca2+ depletion was verified by absence of any contracile response to either 25 mM caffeine or 1 microM inositol 1,4,5-trisphosphate. Then, in the continuous presence of 1 microM calmodulin, bath calcium concentration was raised from zero through 10 microM in half-log increments and the corresponding contractions were recorded. For all permeabilized arteries, the maximum force produced by 10 microM Ca was greater than or equal to that produced by 120 mM potassium-Krebs in the same segment before skinning. The pD2 (-log ED50) values for calcium averaged 6.39 +/- 0.03, 6.77 +/- 0.04, and 6.92 +/- 0.03 in COM, BA, and MCA segments, respectively. In arteries contracted by a constant submaximal concentration of calcium (0.1 microM for BAS and MCA, 0.3 microM for COM), the addition of 5HT produced a dose-dependent and GDPbetaS-sensitive increase in tension of up to 44% maximum. GTPgammaS mimicked the effects of 5HT and prevented further increases in Ca force induced by 5HT. Together, these data demonstrate that cerebrovascular calcium sensitivity is an anatomically heterogenous, physiologically regulated parameter responsive to agonist-induced perturbations.

Animals↗

Mechanisms of platelet-induced angiospastic reactions: potentiation of calcium sensitivity.

Platelet aggregation and adherence to the vessel lumina are common events in various physiological and pathological conditions. However, the mechanisms whereby aggregating platelets cause vasoconstriction remain unclear. We hypothesized that aggregating platelets may alter the Ca2+ sensitivity of the contractile apparatus in smooth muscle cells. We tested this hypothesis using rabbit common carotid arteries permeabilized with beta-escin. In these preparations, the receptor-effector coupling is functionally intact, while the intracellular ionic composition, in particular calcium concentration, may be clamped. Aggregating platelets in the presence of 100 microM GTP left-shifted the pCa-force curve. On average, the pD2 for calcium was 6.30 +/- 0.05 and 6.86 +/- 0.07 in the absence and presence of platelets, respectively (p < 0.01). This increase in calcium sensitivity was prevented by blockade of G-proteins with guanosine 5'-O-(2-thiodiphosphate). Platelets modulated calcium sensitivity at concentrations markedly lower than those that produce contractions of intact vessels. These data suggest that platelet activation in the vicinity of the vessel wall may sensitize the smooth muscle contractile apparatus to calcium via a G-protein-dependent mechanism. This phenomenon may enhance vessel responses to vasoactive substance that produce vessel contraction via elevation of intracellular calcium concentrations.

Animals↗

Effect of chronic hypoxia on myometrial responsiveness in the pregnant rat.

Mechanisms involving the timing of normal parturition are not well understood in most animal species. To gain a greater understanding of the mechanisms, we employed hypoxia to perturb the normal system of parturition. The present study was designed to investigate the effects of chronic hypoxia on myometrial contractility in the near-term pregnant rat. Rats were exposed to room air (control) or to continuous hypoxia (10.5% O2) either from experimental days 19 through 21 (2-day exposure) or from experimental days 15 through 21 (6-day exposure). On day 21, blood was collected for hormone assays, and the uterine horns were collected from each dam. One horn was snap-frozen in liquid nitrogen for oxytocin (OT) receptor analysis, and the other was used for in vitro assessment of myometrial contractile responses to cumulative doses of OT or arginine vasopressin (AVP). Hypoxic exposure resulted in approximately 60% reduction of the maximal myometrial contractile response to OT and a significant reduction in OT binding sites from 256.9 +/- 34.9 to 84.9 +/- 21.3 fmol/mg protein (P<0.01). In contrast, the contractile response to AVP was unaffected after exposure to chronic hypoxia (P> 0.05). Additionally, we observed no difference in the plasma concentrations of estrogen, progesterone, and corticosterone. We conclude that chronic hypoxia decreased the effectiveness of OT-specific contractile mechanisms, at least partially through a decrease in OT binding sites.

Animals↗

NE-induced contraction, alpha 1-adrenergic receptors, and Ins(1,4,5)P3 responses in cerebral arteries.

Adrenergic-mediated responses in cerebral vessels in vitro differ with vessel segment. We performed this study to test the hypothesis that these vessel-specific cerebral artery norepinephrine (NE)-induced contractility changes are mediated in part by differences in alpha 1-adrenergic receptor (alpha 1-R) density (Bmax) or antagonist dissociation constant (KD), and/or inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] synthesis. In common carotid (Com), circle of Willis (Wil), and middle cerebral arteries (MCA) from adult sheep we measured NE-induced contractions. We also quantified alpha 1-R in these, and in anterior, middle, and posterior (AMP) cerebral arteries and cerebral microvessels (Micro). In addition, we quantified NE-induced Ins(1,4,5)P3 synthesis. pD2 values of Com and MCA were 5.2 +/- 0.1 and 6.3 +/- 0.1, respectively. In contrast, the MCA maximum response to NE compared with K+ was much lower than that of the Com. In the Com, Wil, AMP, and Micro, alpha 1-R Bmax was 54 +/- 3, < 5 +/- 2, 23 +/- 3, and 35 +/- 3 fmol/mg protein, respectively. KD averaged 0.20 +/- 0.05 nM in the several vessel groups. In Com and in AMP cerebral arteries, NE produced a rapid increase in Ins(1,4,5)P3 with a peak at 45 s, and 50% effective concentration of 5.5 +/- 0.2 microM. NE stimulated a 240% increase of Ins(1,4,5)P3 in both Com and AMP, whereas Wil showed essentially no response. The ovine MCA was more sensitive to NE than was the Com. In contrast, MCA showed a much lower maximum contractile response to NE compared with K+. Cerebral arteries (AMP) had only about half the alpha 1-R density of the Com. In AMP cerebral arteries, both the basal and NE-stimulated Ins(1,4,5)P3 values were much less than those of the Com. In MCA, the ratio of Ins(1,4,5)P3 response to alpha 1-R Bmax was much greater than in Com. These findings suggest important artery-to-artery differences in components of the cerebrovascular alpha 1-R-mediated contractile pathway. They also suggest considerable potential for modulation of pharmacomechanical coupling and homeostatic regulation of cerebrovascular tone.

Animals↗

Developmental changes in alpha 1-adrenergic receptors, IP3 responses, and NE-induced contraction in cerebral arteries.

Cerebral arteries show significant developmental and artery-specific changes in noradrenergic-mediated contraction. To test the hypothesis that these changes result from differences in the density of alpha 1-adrenergic receptors (alpha 1-ARs) and/or norepinephrine (NE)-induced inositol 1,4,5-trisphosphate [Ins(1,4,5)P3,IP3] synthesis, we quantified these variables and the NE-induced contraction in the common carotid artery (Com) and main branch cerebral arteries (MBC) from term fetal (approximately 140 gestational day) and newborn (2- to 5-day) sheep and compared them with adult values. In fetal and newborn Com, maximal contractions to NE (percent K+ maximum response) were 132 +/- 14 and 118 +/- 9%, respectively (adult = 92 +/- 7%). For fetal and newborn middle cerebral artery, these values were 34 +/- 10 and 43 +/- 7%, respectively (adult = 24 +/- 7%). alpha 1-AR density values in Com of fetal and newborn sheep were 113 +/- 18 and 106 +/- 4 fmol/mg protein, respectively (adult = 54 +/- 3 fmol/mg protein). For the MBC, density values were 47 +/- 2 and 24 +/- 3 fmol/mg protein, respectively (adult = 23 +/- 3 fmol/protein). In term fetal and newborn MBC, NE produced dose-dependent increases in Ins(1,4,5)P3, the maximal increases above basal values being 245 +/- 40 and 189 +/- 16%, respectively (adult = 254 +/- 35%). Neither fetus nor newborn Com showed significant Ins(1,4,5)P3 responses to NE. We concluded that in fetal and newborn Com and MBC, alpha 1-AR density and NE-induced Ins(1,4,5)P3 response varied as a function of developmental age and specific vessel. However, these variations did not correlate with NE-induced maximum contraction. Thus we reject the hypothesis that age-dependent and vessel-specific differences of cerebral artery adrenergic-mediated contraction are a function of alpha 1-AR density or Ins(1,4,5)P3 response. Rather, the differences would appear to result from other factors such as non-Ins(1,4,5)P3-mediated calcium activation and/or sensitivity to Ins(1,4,5)P3. The studies also suggest considerable potential for maturational modulation of pharmacomechanical coupling and homeostatic regulation of cerebrovascular tone.

Aging↗

Effects of maturation on cyclic GMP metabolism in ovine carotid arteries.

Previous studies suggest that elevated basal levels of cGMP in newborn arteries may help explain why vascular resistance is lower in newborns than adults. To explore the reasons why basal cGMP is higher in neonatal arteries, the present studies examined rates of cGMP synthesis and degradation in newborn and adult ovine common carotid arteries. The measurements were performed in both intact and homogenized arteries, and results were normalized relative to cell water to estimate intracellular concentrations and minimize errors due to compositional differences between newborn and adult arteries. Steady state levels of cGMP measured under baseline conditions averaged 0.11 +/- 0.02 microM in adult arteries and 0.59 +/- 0.11 microM in newborn arteries. These resting cGMP levels were unaffected by endothelium removal. Under baseline conditions, steady state rates of cGMP synthesis (mumol of cGMP/L of cell water/min) were higher in newborn (0.31 +/- 0.06) than in adult (0.15 +/- 0.04) arteries. Maximal rates of cGMP degradation (mumol of cGMP/L of cell water/min) measured in artery homogenates were also much higher in preparations of newborn (106 +/- 6) than of adult (78 +/- 6) arteries. Together, these data suggest that the reason resting cGMP concentrations were higher in newborn than in adult arteries was due at least in part to a higher basal rate of cGMP synthesis in the newborn. Estimates of apparent Km values for PDE were also greater in newborn (2.9 microM) than in adult (1.5 microM) preparations, suggesting that age-related differences in the Km for PDE may also contribute to the elevated basal concentration of cGMP observed in the newborn.

1-Methyl-3-isobutylxanthine↗

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↗