Effect of nitroprusside on regional cerebral cyclic GMP, blood flow and O2 consumption in rat

This investigation was conducted to test whether topical nitroprusside (NP), a cytosolic guanylate cyclase activator, would increase the level of cyclic GMP and alter O₂ consumption or blood flow in the cerebral cortex of rats. Male Long-Evans rats were used in a control (n = 9), low dose NP (n = 13, 10⁻³ M) or high dose NP (n = 12, 10⁻² M) group. Nitroprusside or saline was topically applied to the right side of the cerebral cortex and the left side was used as a control. The cyclic GMP level was determined in five rats in each group using a radioimmunoassay. In the o ther rats in each group, regional cerebral blood flow was measured by [¹⁴C]iodoantipyrine and regional arterial and venous O₂ saturations were determined microspectrophotometrically. Nitroprusside significantly increased the cyclic GMP level from 21.4 ± 12.0 pmol/g (contralateral cortex) to 52.2 ± 36.7 pmol/g (NP treated cortex) in low dose nitroprusside group and from 19.9 ± 22.6 pmol/g (contralateral cortex) to 58.5 ± 15.1 pmol/g (NP treated cortex) in high dose nitroprusside group. High dose nitroprusside significantly increased cerebral blood flow from 80 ± 11 ml · min⁻¹ · 100 g (contralateral cortex) to 114 ± 11 ml · min⁻¹ · 100 g (NP treated cortex). However, there was no significant difference in O₂ extraction and O₂ consumption between the NP treated cortex and contralateral cortex in either the low or the high dose NP groups. In the high dose NP group, the O₂ extraction was 8.0 ± 1.3 ml O₂ · 100 ml⁻¹ in the treated cortex and 8.8 ± 1.5 ml O₂ · 100 ml⁻¹ in the contralateral cortex, while the O₂ consumptions in the NP treated cortex and contralateral cortex were 8.1 ± 1.3 ml O₂ · min⁻¹ · 100 g⁻¹ and 7.3 ± 1.2, respectively. Thus, NP increased the cyclic GMP level without a significant change in O₂ consumption in the cerebral cortex. Our data suggested that O₂ consumption in the cerebral cortex was not affected by the increased level of cyclic GMP.     

17Beta-estradiol blocks NMDA-induced increases in regional cerebral O(2) consumption

We tested the hypothesis that 17beta-estradiol would reduce the cerebral O(2) consumption response to stimulation of N-methyl-D-aspartate (NMDA) receptors. We determined NMDA receptor density in 10 ovariectomized Wistar female rats equally divided into a control group and 17beta-estradiol (500 microg/21 days) treated group. An autoradiographic assay using 125I-MK-801, an NMDA antagonist, was used to measure specific binding to NMDA receptors. Another 14 ovariectomized rats were separated into 17beta-estradiol and control groups to determine cerebral blood flow (14C-iodoantipyrine) and O(2) consumption (microspectrophotometry). 17Beta-estradiol caused a 20% decrease in specific binding to cortical NMDA receptors. After topical cortical stimulation with 10(-3)M and 10(-4)M NMDA, blood flow increased significantly in control from 73+/-5 in the saline treated cortex to 110+/-8 ml/min/100 g with 10(-3)M NMDA. In contrast, there was no significant change in blood flow in the 17beta-estradiol treated animals. Cerebral O(2) extraction increased significantly in the 10(-3)M NMDA treated cortex in both groups. Cerebral O(2) consumption in the control group significantly increased by 53%, from 3.7+/-0.2 to 5.7+/-0.5 with 10(-4)M NMDA and 72% to 6.4+/-2.4 ml O(2)/min/100 g with 10(-3)M NMDA. The 17beta-estradiol group demonstrated no significant difference between the saline treated and NMDA treated cortex. Thus, 17beta-estradiol blocked the effects of NMDA on cerebral O(2) consumption and this was associated with a slightly decreased number of NMDA receptors.     

Cerebral O2 consumption in young Eker rats, effects of GABA blockade: implications for autism

Since there is a strong correlation between tuberous sclerosis and autism, we used a tuberous sclerosis model (Eker rat) to test the hypothesis that the increased regional cerebral O₂ consumption in the Eker rat might be associated with autism. We also examined whether this increased cerebral O₂ consumption was related to changes in the activity of the gamma-aminobutyric acid (GABA) inhibitory system. Young (4 weeks) male control Long Evans (n = 14) and Eker (n = 14) rats (70–100 g) were divided into control and bicuculline (1 mg/kg/min for 2 min then 0.1 mg/kg/min for 13 min, GABA_A receptor antagonist) treated animals. Cerebral regional blood flow (¹⁴C-iodoantipyrine) and O₂ consumption (cryomicrospectrophotometry) were determined in isoflurane anesthetized rats. We found significantly increased basal O₂ consumption in the cortex (6.3 ± 0.7 ml O₂/min/100 g Eker vs. 5.1 ± 0.2 ml O₂/min/100 g control), hippocampus and cerebellum, but not the pons. Regional cerebral blood flow was also elevated in the cortex and hippocampus in Eker rats at baseline, but cerebral O₂ extractions were similar. Bicuculline significantly increased O₂ consumption in the cortex (6.5 ± 0.3) and all other regions of the control rats, but had no effect on cortex (5.9 ± 1.5) or other regions of the Eker rats. Cerebral blood flow followed a similar pattern. In conclusion, Eker rats had significantly elevated cerebral O₂ consumption and blood flow, but this was not affected by GABA receptor blockade. This suggested a reduced activity of the GABA_A receptor in the brains of Eker rats. This may have important implications in the treatment of autism.     

Effects of aging on N-methyl-d-aspartate (NMDA)-induced GnRH and LH release in female rats

In order to evaluate if the changes of the hypothalamic-pituitary-ovary axis that induce a decrease in fertility and modifications in the sexual cycles during senescence involve modifications in the regulatory action of excitatory amino acid neurotransmission on GnRH neurons, we measured the in vitro effects of NMDA on GnRH release by the anterior preoptic and medial basal hypothalamic areas (APOA-MBH) of castrated aging (18 months old) and young (90 days of age) rats. In a second series of experiments the in vivo LH release response to intrahypothalamic (push-pull) administration of NMDA to aged and young castrated female rats was also determined. A similar rate of basal GnRH release was observed in old and young rats during the incubation time. The addition of NMDA to the medium significantly increased GnRH release in both groups; nevertheless, the GnRH release response to NMDA was significantly lower in old ( P < 0.01) than in young rats (Young: Basal: 50 ± 10; NMDA 15′: 410 ± 63, 22,5′: 1,469 ± 300; Old; Basal: 47±10 NMDA 15′:210 ± 30; 22,5′: 350 ± 65 ng/GnRH/mg . protein). The LH levels measured throughout the in vivo experiments indicated that basal LH concentrations were significantly lower in the aged group. The mean LH concentrations (fractions 1 to 6) was significantly lower in the aged group (Young: 3.9 ± 0.07, Old: 2.4 ± 0.03 ng/ml, P < 0.01). The LH release response to NMDA measured 10 min after the intrahypothalamic administration of the glutamate agonist was significantly lower in aged rats (4 .2 ± 1.6 ng/ml) as compared to young animals (18.0 ± 6.1 ng/ml; P < 0.05). LH levels in young rats increased to 580% vs., and only 47% in aged rats as compared to previous basal values. In conclusion, present results demonstrate that the GnRH responses to NMDA neurotransmission, which has a predominantly excitatory effects on GnRH neurons, is significantly decreased in old rats, these data give further support to the hypothesis that a decrease in the excitatory inputs to GnRH neurons could be directly involved in the reduction of the hypothalamic-pituitary-ovary axis activity observed during aging.     

PhTx4, a new class of toxins from Phoneutria nigriventer spider venom, inhibits the glutamate uptake in rat brain synaptosomes

We report the characterization of a new class of glutamate uptake inhibitors isolated from Phoneutria nigriventer venom. Glutamate transport activity was assayed in rat cerebrocortical synaptosomes by using [³H]- -glutamate. PhTx4 inhibited glutamate uptake in a dose dependent manner. The IC₅₀ value obtained was 2.35±0.9 μg/ml which is in the observed range reported for glutamate uptake blockers. Tx4-7, one of PhTx4 toxins, showed the strongest inhibitory activity (50.3±0.69%, n=3).     

Effects of hyperosmolar mannitol on regional oxygen supply and consumption in the newborn pig

Abstract

Previous work indicated that opening the blood-brain barrier with hyperosmotic mannitol decreased local venous O₂ saturation and increased cerebral O₂ consumption. This study was performed to assess the vascular effect of hypertonic mannitol on oxygen supply/consumption balance in the newborn pig and to determine the role of nitric oxide in mediating the effects of mannitol. Animals were anesthetized with (J.chloralose and mechanically ventilated to maintain their blood gases within normal range. Retrograde catheterization of the right carotid artery was performed to inject 12 ml to 25% mannitol over a 30 sec interval. In one group of animals (n = 5), the blood-brain barrier transfer coefficient (Ki) to ¹⁴C-a aminoisobutyric acid or 14C-urea (n = 4) was measured 72 min after mannitol. In another group of animals (n = 9), regional cerebral blood flow and small vein O₂ saturation was measured using 14C-iodoantripyrine and microspectrophotometry. Similar measurements were made in other groups of animals (n = 9) after pretreatment with 10 mg kg^-1 i.v. of N-omega-nitro-L-arginine methyl ester (L-NAME), 20 min before mannitol injection. The mannitol injection did not increase Ki or local cerebral O₂ consumption. It resulted in a decreased small vein O₂ saturation in the ipsilateral cortex (46 ± 3%) in comparison to the contralateral cortex (55 ± 2%). The O₂ supply/consumption ratio decreased in the ipsilateral cortex in the mannitol injected animals (2.14 ± 0.23) in comparison to the contralateral cortex (2.76 ± 0.28). Pretreatment with L-NAME abolished this effect of mannitol (small vein O₂ saturation 59 ± 2% in ipsilateral cortex and 58 ± 2% in the contralateral cortex; O₂ supply/consumption 2.68 ± 0.17 in the ipsilateral cortex and 2.65 ± 0.16 in the contralateral cortex). We conclude that hypertonic mannitol adversely affects O₂ supply/consumption balance, without increasing bloQd-brain barrier transport, and this effect is blocked by L-NAME, a nitric oxide synthase antagonist. [Neural Res 1997; 19: 204–210]     

Increased cerebral oxygen consumption in Eker rats and effects of N-methyl-D-aspartate blockade: Implications for autism

Because there is a strong correlation between tuberous sclerosis and autism, we used a tuberous sclerosis model (Eker rat) to test the hypothesis that these animals would have an altered regional cerebral O2 consumption that might be associated with autism. We also examined whether the altered cerebral O2 consumption was related to changes in the importance of N-methyl-D-aspartate (NMDA) receptors. Young (4 weeks) male control Long Evans (N = 14) and Eker (N = 14) rats (70-100 g) were divided into control and CGS-19755 (10 mg/kg, competitive NMDA antagonist)-treated animals. Cerebral regional blood flow (14C-iodoantipyrine) and O2 consumption (cryomicrospectrophotometry) were determined in isoflurane-anesthetized rats. NMDA receptor protein levels were determined by Western immunoblotting. We found significantly increased basal O2 consumption in the cortex (6.2 +/- 0.6 ml O2/min/100 g Eker vs. 4.7 +/- 0.4 Long Evans), hippocampus, cerebellum, and pons. Regional cerebral blood flow was also elevated in Eker rats at baseline, but cerebral O2 extraction was similar. CGS-19755 significantly lowered O2 consumption in the cortex (2.8 +/- 0.3), hippocampus, and pons of the Long Evans rats but had no effect on cortex (5.8 +/- 0.8) or other regions of the Eker rats. Cerebral blood flow followed a similar pattern. NMDA receptor protein levels (NR1 subunit) were similar between groups. In conclusion, Eker rats had significantly elevated cerebral O2 consumption and blood flow, but this was not related to NMDA receptor activation. In fact, the importance of NMDA receptors in the control of basal cerebral O2 consumption was reduced. This might have important implications in the treatment of autism.     

Effects of perfusion flow rate, prostaglandin F2α, phenylephrine, and serotonin on isolated, perfused brains of spontaneously hypertensive rats

Effects of perfusion flow rate and three vasoconstrictors, phenylephrine, prostaglandin F_2α (PGF_2α) and serotonin, on isolated, perfused brain preparations of spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto rats (WKY) were investigated. The basal perfusion pressure of the cerebral vascular beds at a flow rate of 2.5 ml/min was 48 ± 3mm Hg(n = 11) in SHR and 32 ± 2mm Hg(n = 12)in WKY(P < 0.005). The perfusion pressures at all flow rates tested (2.5−6.5 ml/min) in SHR were significantly greater than those in WKY. Concentration-perfusion pressure curves for the vasoconstrictors showed that the brain vascular bed was much more reactive to sertonin compared with phenylephrine and PGF_2α. EC₅₀ values (−logM) for serotonin in the perfused brains of SHR and WKY were 7.0 ± 0.06 (n = 10)and6.5 ± 0.06 (n = 11), respective (P < 0.01). There were no differences in EC₅₀ values for phenylephrine or PGF_2α between SHR and WKY. Exogenous serotonin and phenulephrine caused significantly greater maximal vasoconstrictor responses in SHR compared with WKY, while the pressor response to PGF_2α was very weak and no significant difference between SHR and WKY preparations was observed. These results indicate that cerebral vascular beds in SHR exhibit higher cerebrovascular resistance than those in WKY, and that reactivity and sensitivity to serotonin and reactivity to phenylephrine in SHR rats are enhanced to a greater extent compared to WKY.     

Preservation of autoregulatory cerebral vasodilator responses to hypotension after inhibition of nitric oxide synthesis

Effects of inhibition of nitric oxide (NO) synthesis on the cerebrovascular autoregulatory vasodilator response to hypotension were studied in conscious rats. Cerebral blood flow (CBF) was determined with [¹⁴C]iodoantipyrine in a saline-treated control group and in three groups following inhibition of NO synthase activity by twice daily intraperitoneal injections of 50 mg/kg ofN^G-nitro-l-arginine methyl ester (l-NAME) for four days. In the saline-control group (n = 8) and in thel-NAME-treated Group (a) (n = 8) CBF was determined while systemic mean arterial blood pressure (MABP) remained at its resting level (means ± S.D., 128±6 and 151±11 mmHg, respectively). In the other groups CBF was determined after MABP was reduced by blood withdrawal to 118±9 and 88±8 mmHg in Groups (b) (n = 8) and (c) (n = 8), respectively. Despite the elevated MABP, global CBF was significantly lower inl-NAME-treated Group (a) than in the saline-controls (P < 0.005), indicating cerebral vasoconstriction striction resulting from inhibition of NO synthesis. Global CBF was not significantly reduced further in the two groups with hypotension. Local CBF in the hypotensive rats showed no significant reductions below values inl-NAME-treated control rats (Group (a)) in 31 of 32 brain structures; the only exception was in the auditory cortex of the severely hypotensive rats (Group (c)). The autoregulatory mechanism for cerebral vasodilatation to compensate for reduced arterial blood pressure is maintained following inhibition of NO synthesis.     

l-Arginine ameliorates cerebral blood flow and metabolism and decreases infarct volume in rats with cerebral ischemia

Effects ofl-arginine, 300 mg/kg, i.p., on the regional cerebral blood flow (rCBF), brain metabolism, and infarct volume were examined in spontaneously hypertensive rats subjected to occlusion of both left middle cerebral artery and left common carotid artery. Rats treated withl-arginine had higher rCBF, determined by hydrogen clearance method, in the ischemic core (7 ± 1 ml/100 g/min, mean ± S.E.M.) and penumbral regions (16 ± 2) than did rats treated with saline (5 ± 0 and 7 ± 1, respectively). Simultaneously,l-arginine attenuated metabolic derangement in the ischemic tissue at 60 min, i.e. well maintained adenosine triphosphate (ATP) in ischemic region (1.29 ± 0.07 mmol/kg inl-arginine group vs. 1.05 ± 0.06 in saline group), and also close to normal levels in ATP (2.61 ± 0.02 mmol/kg vs. 2.45 ± 0.05), glucose (2.29 ± 0.12 mmol/kg vs. 1.80 ± 0.17) and lactate (1.63 ± 0.10 mmol/kg vs. 2.24 ± 0.21) in periischemic region. In another experiment, the effects ofl-arginine on rCBF in the subcortical regions and on infarct volume were evaluated.l-arginine, compared with saline, increased rCBF by 8 ml/100 g/min in the ischemic side and reduced infarct volume by 29% at 24 h of ischemia. These findings support thatl-arginine may be potentially useful for the treatment of acute cerebral ischemia.     

Role of nitric oxide in regulation of cerebral microvascular tone and autoregulation of cerebral blood flow in cats

The role of nitric oxide in the regulation of cerebrocortical microvascular tone and autoregulation of cerebral blood flow (CBF) was examined in 24 anesthetized cats. The local cerebral blood volume (CBV), mean transit time of blood (MTT), and CBF in the cortex were measured by our photoelectric method. CBV represents the cumulative dimensions of the cerebral microvessels. Intravenous injection of 0.35–0.7 mg/kg/minN^G-monomethyl-l-arginine (l-NMMA), an inhibitor of nitric oxide synthesis, significantly increased mean arterial blood pressure (MABP; 8.4–14.1%,P < 0.01), decreased CBV (15.2–28.7%,P < 0.01), and decreased CBF (20.0–29.8%,P < 0.01) in a dose-related manner. The changes in MABP, CBV, and CBF elicited byl-NMMA were inhibited (P < 0.05) by simultaneous infusion of 35 mg/kg/minl-arginine. Autoregulation of CBF was examined during controlled hypotension of −30 to −40 mmHg (artificial bleeding) and recovery of blood pressure (reinfusion of blood). Although CBF remained constant with blood pressure changes in the control state (ΔCBF/ΔMABP of 0.037±0.155 with hypotension), CBF became dependent on blood pressure changes (ΔCBF/ΔMABP of 0.478±0.135, P < 0.05) during infusion of 0.35 mg/kg/minl-NMMA. It is concluded that nitric oxide participates in both the regulation of basal tone of cerebral microvessels and the autoregulation of CBF.     

Effect of upregulation of AMPA glutamate receptors on cerebral O2 consumption and blood flow in rat

AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate) receptors, in cerebral cortex, underwent upregulation (35% increase) following chronic blockade with a non-competitive AMPA receptor antagonist, GYKI 52466 (1-(aminophenyl)-4-methyl-7, 8-methylenedioxy-5H-2,3-benzodiazepine). Such upregulation did not alter basal cerebrocortical blood flow or O(2) consumption. There was a much higher increase in blood flow and O(2) consumption in the upregulated, agonist (AMPA) stimulated cortices of anesthetized rats.     

D1 dopamine and NMDA receptors interactions in the medial prefrontal cortex: Modulation of spatial working memory in rats

Dopamine (DA) and N-methyl-d-aspartate (NMDA) receptors seem to be critically involved in working memory processing in the medial prefrontal cortex (mPFC). Effects of NMDA receptors blockade on dopamine D₁ receptors activation in the mPFC on spatial working memory was investigated. Adult male Wistar rats, well trained in an eight-arm radial maze and bilaterally cannulated in the mPFC, received intracortical administrations of saline (SAL) or SKF-38393 (DA D₁ receptor agonist) followed, 10 min later, by MK-801 (non-competitive NMDA receptor antagonist). They were tested in 1 h delayed tasks after 5 min of the second administration. SKF-38393 (0.56 and 1.8 μg) was disruptive to working memory, increasing significantly the number of errors in the 1 h post-delay performance when administered into the mPFC. MK-801, at doses with no significant effects alone (0.32 or 1.0 μg), reduced significantly the disruptive effect of 0.56 μg SKF-38393. These results showed that the disruptive effect of DA D₁ receptors activation in the mPFC on working memory was significantly reduced by an open-channel NMDA receptor blockade, suggesting that the processing of working memory in the mPFC involving DA D₁ receptors depend, at least in part, of NMDA receptors activity in this cortical area.     

Decrease in GABA synthesis rate in rat cortex following GABA-transaminase inhibition correlates with the decrease in GAD67 protein

γ-Aminobutyric acid (GABA) synthesis in the brain is mediated by two major isoforms of glutamic acid decarboxylase, GAD₆₅ and GAD₆₇. The contribution of these isoforms to GABA synthesis flux (V_GAD) is not known quantitatively. In the present study we compared V_GAD in cortex of control and vigabatrin-treated rats under α-chloralose/70% nitrous oxide anesthesia, with total GAD activity and GAD isoform composition (GAD₆₅ and GAD₆₇) measured by enzymatic assay and quantitative immunoblotting. V_GAD was determined by re-analysis of ¹³C NMR data obtained ex vivo and in vivo during infusions of [1-¹³C]glucose using an extension of a model of glutamate–glutamine cycling that included a discrete GABAergic neuronal compartment with relevant interconnecting fluxes. V_GAD was significantly lower in vigabatrin-treated rats (0.030–0.05 μmol/min per g, P<0.003) compared to the non-treated control group (0.10–0.15 μmol/min per g). The 67–70% decrease in V_GAD was associated with a 13% decrease in total GAD activity (P=0.01) and a selective 44±15% decrease in GAD₆₇ protein (from 0.63±0.10 to 0.35±0.08 μg protein/mg tissue, P<0.05); GAD₆₅ protein was unchanged. The reduction in GAD₆₇ protein could account for a maximum of 65% of the decrease in V_GAD in vigabatrin-treated animals suggesting that inhibition of GAD₆₅ must have also occurred in these experiments, although product inhibition of GAD₆₇ by increased GABA could play a role. GAD₆₇ could account for 56–85% of cortical GABA synthesis flux under basal conditions and the entire flux after vigabatrin treatment.     

Cerebral oxygen metabolism of rats using injectable 15O-oxygen with a steady-state method

To develop a less-stressful and simple method for measurement of the cerebral metabolic rate of oxygen (CMRO₂) in small animals, the steady-state method was applied to injectable ¹⁵O₂-PET (¹⁵O₂-positron emission tomography) using hemoglobin-containing vesicles (¹⁵O₂-HbV). Ten normal rats and 10 with middle cerebral arterial occlusion (MCAO) were studied using a small animal PET scanner. A series of ¹⁵O-PET scans with C¹⁵O-labeled HbV, H₂¹⁵O, and ¹⁵O₂-HbV were performed with 10 to 15 minutes intervals to measure cerebral blood volume (CBV), cerebral blood flow (CBF), and CMRO₂. Positron emission tomography scans were started with a tracer injection using a multiprogramming syringe pump, which provides a slowly increasing injection volume to achieve steady-state radioactivity for H₂¹⁵O and ¹⁵O₂-HbV scans. The radioactivity concentration of ¹⁵O rapidly achieved equilibrium in the blood and whole brain at about 2 minutes after H₂¹⁵O and ¹⁵O₂-HbV administration, which was stable during the scans. The whole brain mean values of CBF, CBV, and CMRO₂ were 54.3±2.0 mL per 100 g per minute, 4.9±0.4 mL/100 g, and 2.8±0.2 μmoL per g per minute (6.2±0.4 mL per 100 g per minute) in the normal rats, respectively. In the MCAO model rats, all hemodynamic parameters of the infarction area on the occlusion side significantly decreased. The steady-state method with ¹⁵O-labeled HbV is simple and useful to analyze hemodynamic changes in studies with model animals.     

Lesions of the rostral ventrolateral medulla reduce the cerebrovascular response to hypoxia

Sympathoexcitatory neurons of the rostral ventrolateral medulla are tonically active and required for maintenance of resting levels of arterial pressure. They are also selectively excited by hypoxia and responsible for the associated sympathoexcitation. Since electrical or chemical stimulation of RVL will increase regional cerebral blood flow (rCBF) independently of changes in regional cerebral glucose utilization (rCGU) we investigated whether the RVL was also required to maintain resting levels of rCBF and also participated in the cerebrovascular vasodilation elicited by hypoxia. Rats were anesthetized (chloralose; 40 mg/kg, s.c.), paralyzed (tubocurarine) and ventilated (100% O₂). rCBF was measured in 10 dissected brain regions using [¹⁴C]iodoantipyrine; rCGU was measured by 2-deoxy-d-[¹⁴C]glucose. in controls (n = 6) rCBF ranged56 ± 5 in corpus callosum to101 ± 6ml/min× 100g in inferior colliculus. Hypoxic-hypoxia (PaO₂ - 36 ± 1mmHg, n = 6) increased rCBF in all structures maximally, at 204% of control, in occipital cortex. Hypercapnia (PaCO₂ = 63.5 ± 0.9, n =5) also increased rCBFP < 0.01) maximally to 199% of control in superior colliculus, Spinal cord transection with maintenance of arterial pressure did not affect resting rCBF and increased the vasodilation to hypoxia (PaO₂ = 39 ± 1mmHg, n = 5) from 2- to 3-fold in all structuresP < 0.01). Bilateral lesions within the RVL had no effect on resting rCBF or rCGU. However, they significantly reduced, in all areas by 50–69% (P < 0.01, n = 5), the cerebrovascular dilation elicited by hypoxia but not hypercapnia, Bilateral lesions in the spinal trigeminal nucleus (PaO₂ = 35 ± 1; n = 6), or transection of the IXth and Xth cranical nerves did not affect the rCBF response to hypoxia(PaO₂ = 41 ± 2; n = 6) (P > 0.05) indicating that the effect of RVL Lesions was not attributable to interference with arterial baro- or chemoreceptor reflexes. We conclude that neurons within RVL are not responsible for maintaining tonic levels of rCBF. However they contribute to the cerebrovascular vasodilation elicited by hypoxia but not hypercapnia. The cerebrovascular response to hypoxia appears reflexive and, in part, due to stimulation of oxygen-sensing neurons in RVL. In contrast, the vasodilation elicited by hypercapnia reflects local chemical signals in the cerebral microcirculation     

Influence of 100% and 40% oxygen on penumbral blood flow,oxygen level,and T2*-weighted MRI in a rat stroke model

Accurate imaging of the ischemic penumbra is a prerequisite for acute clinical stroke research. T₂^* magnetic resonance imaging (MRI) combined with an oxygen challenge (OC) is being developed to detect penumbra based on changes in blood deoxyhemoglobin. However, inducing OC with 100% O₂ induces sinus artefacts on human scans and influences cerebral blood flow (CBF), which can affect T₂^* signal. Therefore, we investigated replacing 100% O₂ OC with 40% O₂ OC (5 minutes 40% O₂ versus 100% O₂) and determined the effects on blood pressure (BP), CBF, tissue p₂, and T₂^* signal change in presumed penumbra in a rat stroke model. Probes implanted into penumbra and contralateral cortex simultaneously recorded p₂ and CBF during 40% O₂ (n=6) or 100% O₂ (n=8) OC. In a separate MRI study, T₂^* signal change to 40% O₂ (n=6) and 100% O₂ (n=5) OC was compared. Oxygen challenge (40% and 100% O₂) increased BP by 8.2% and 18.1%, penumbra CBF by 5% and 15%, and penumbra p₂ levels by 80% and 144%, respectively. T₂^* signal significantly increased by 4.56%±1.61% and 8.65%±3.66% in penumbra compared with 2.98%±1.56% and 2.79%±0.66% in contralateral cortex and 1.09%±0.82% and −0.32%±0.67% in ischemic core, respectively. For diagnostic imaging, 40% O₂ OC could provide sufficient T₂^* signal change to detect penumbra with limited influence in BP and CBF.     

Role of cAMP and K channel-dependent mechanisms in piglet hypoxic/ischemic impaired nociceptin/orphanin FQ-induced cerebrovasodilation

This study was designed to determine the role of altered cAMP and K⁺ channel-dependent mechanisms in impaired pial artery dilation to the newly described opioid, nociceptin/orphanin FQ (NOC/oFQ) following hypoxia/ischemia in newborn pigs equipped with a closed cranial window. Recent studies have observed that NOC/oFQ elicits pial dilation via release of cAMP, which, in turn, activates the calcium sensitive (K_ca) and the ATP-dependent K⁺ (K_ATP) channel. Global cerebral ischemia (20 min) was induced via elevation of intracranial pressure, while hypoxia (10 min) decreased pO₂ to 35±3 mmHg with unchanged pCO₂. Topical NOC/oFQ (10⁻⁸, 10⁻⁶ M) induced vasodilation was attenuated by ischemia/reperfusion (I+R) and reversed to vasoconstriction by hypoxia/ischemia/reperfusion (H+I+R) at 1 h of reperfusion (control, 9±1 and 16±1%; I+R, 3±1 and 6±1%; H+I+R, −7±1 and −12±1%). Such altered dilation returned to control values within 4 h in I+R animals and within 12 h in H+I+R animals. NOC/oFQ dilation was associated with elevated CSF cAMP in control animals but such biochemical changes were attenuated in I+R animals and reversed to decreases in cAMP concentration in H+I+R animals (control, 1037±58 and 1919±209 fmol/ml; I+R, 1068±33 and 1289±30 fmol/ml; H+I+R, 976±36 and 772±27 fmol/ml for absence and presence of NOC/oFQ 10⁻⁶ M, respectively). Topical 8-Bromo cAMP (10⁻⁸, 10⁻⁶ M) pial dilation was unchanged by I+R but blunted by H+I+R (control, 10±1 and 20±1%; I+R, 11±1 and 20±2%; H+I+R, 0±1 and 0±2%). Pituitary adenylate cyclase activating polypeptide and cromakalim, adenylate cyclase and K_ATP channel activators, respectively, elicited dilation that was blunted by both I+R and H+I+R while NS1619, a K_ca channel activator, elicited dilation that was unchanged by I+R but blunted by H+I+R. These data indicate that impaired NOC/oFQ dilation following I+R results form altered adenylate cyclase and K_ATP channel-dependent mechanisms. These data further indicate that impaired NOC/oFQ dilation following H+I+R results not only from altered adenylate cyclase and K_ATP channel but also from altered cAMP and K_ca channel-dependent mechanisms.     

Mathematical model of oxygen transport in the cerebral cortex

To estimate the magnitude of hyperemia necessary to support oxidative metabolism in the cerebral cortex during functional activation, a mathematical model of O₂ transport from capillary to tissue was developed. Radial and axial gradients of O₂ pressure in tissue surrounding a single capillary were calculated at normal and increased cerebral metabolic rates for O₂. Cone-shaped tissue geometry and nonlinear oxyhemoglobin dissociation were assumed. Local O₂ consumption was assumed to be supported with local tissue pO₂ greater than 1 mmHg. The distribution of tissue pO₂ was also calculated during moderate hypoxemia (p_aO₂=42 mmHg), using experimental values of red blood cell velocity measured in individual capillaries of the rat cerebral cortex using intravital video-microscopy. The model predicted that moderate increases (≤50%) in cerebral O₂ consumption were supported by proportional increases in capillary blood flow. Large increases in O₂ consumption (50–110%) were supported by disproportional increases in flow. During moderate hypoxemia, average tissue pO₂ decreased but oxygen utilization was sustained when capillary flow was increased to a level measured in experiments. The results suggest a proportional relationship between cerebrocortical blood flow and oxygen consumption in the normal physiological range of functional activation.     

Cerebrovascular consequences of altering serotonergic transmission in conscious rat

Many therapeutic strategies aim at altering serotonin brain levels. However, serotonin (5-HT) is known to influence the cerebral circulation. The purpose of this study was to determine the effects of acutely decreasing intracerebral serotonin release upon cerebral blood flow and cerebrovascular reactivity to hypercapnia in conscious rats. To this end, (1) we analyzed the time-course of cortical blood flow changes measured with laser-Doppler flowmetry following injection of 0.1 mg kg⁻¹ 8-OHDPAT (5-HT_1A agonist), and (2) we evaluated the cerebrovascular reactivity to hypercapnia using a quantitative multiregional diffusible tracer technique 5 and 60 min following 8-OHDPAT administration. 8-OHDPAT induced a rapid and transient increase in cortical blood flow (+34%) that was prevented totally by WAY100135 (5-HT_1A antagonist) pre-treatment. Five min following 8-OHDPAT administration, the cerebrovascular responsiveness to hypercapnia was increased significantly in striatum (+27%) and fronto-parietal cortex (+61%). This result is consistent with a vasoconstrictor role of the serotonergic system that becomes manifest during hyperemic conditions.