Brain pHi Research Articles

D-galactose-induced aging in rats - The effect of metformin on bioenergetics of brain, skeletal muscle and liver.

Chronic D-galactose administration induces accelerated aging in rodents. The aim of the study was to find by in vivo31P MRS suitable markers of early stages of brain degeneration on this metabolic model in rats. Additionally, we studied the therapeutic effect of antidiabetic drug metformin. The study has been extended by in vitro determination of mitochondrial function in brain, skeletal muscle and liver mitochondria, oxidative stress parameter thiobarbituric acid reactive substances (TBARS), and lipophilic antioxidants levels. In vivo31P MRS revealed lower intracellular pH (pHi) and lower inorganic phosphate to ATP ratio (Pi/ATP), with higher index of oxidative phosphorylation - phosphocreatine (PCr) to Pi ratio - in brain of rats chronically administered with D-galactose. The function of brain mitochondria was not affected. Administration of metformin diminished changes in brain pHi and plasma TBARS. The function of skeletal muscle mitochondria and their coenzyme Q (CoQ) content were considerably reduced after D-galactose administration. Metformin administered simultaneously with D-galactose did not prevent these changes. The results of in vivo31P MRS revealed evidence of early stage of neurodegeneration that may indicate pre-inflammation. Our data show different susceptibility of brain, skeletal muscle, and liver to the chronic exposure to D-galactose and metformin. The D-galactose model presented in the literature as a model for "age-related dementia" had much more devastating effects on skeletal muscle than on the brain.

Altered brain ion gradients following compensation for elevated CO2 are linked to behavioural alterations in a coral reef fish.

Neurosensory and behavioural disruptions are some of the most consistently reported responses upon exposure to ocean acidification-relevant CO2 levels, especially in coral reef fishes. The underlying cause of these disruptions is thought to be altered current across the GABAA receptor in neuronal cells due to changes in ion gradients (HCO3− and/or Cl−) that occur in the body following compensation for elevated ambient CO2. Despite these widely-documented behavioural disruptions, the present study is the first to pair a behavioural assay with measurements of relevant intracellular and extracellular acid-base parameters in a coral reef fish exposed to elevated CO2. Spiny damselfish (Acanthochromis polyacanthus) exposed to 1900 μatm CO2 for 4 days exhibited significantly increased intracellular and extracellular HCO3− concentrations and elevated brain pHi compared to control fish, providing evidence of CO2 compensation. As expected, high CO2 exposed damselfish spent significantly more time in a chemical alarm cue (CAC) than control fish, supporting a potential link between behavioural disruption and CO2 compensation. Using HCO3− measurements from the damselfish, the reversal potential for GABAA (EGABA) was calculated, illustrating that biophysical properties of the brain during CO2 compensation could change GABAA receptor function and account for the behavioural disturbances noted during exposure to elevated CO2.

Na+/H+ Exchangers and Intracellular pH in Perinatal Brain Injury

Encephalopathy consequent on perinatal hypoxia–ischemia occurs in 1–3 per 1,000 term births in the UK and frequently leads to serious and tragic consequences that devastate lives and families, with huge financial burdens for society. Although the recent introduction of cooling represents a significant advance, only 40 % survive with normal neurodevelopmental function. There is thus a significant unmet need for novel, safe, and effective therapies to optimize brain protection following brain injury around birth. The Na+/H+ exchanger (NHE) is a membrane protein present in many mammalian cell types. It is involved in regulating intracellular pH and cell volume. NHE1 is the most abundant isoform in the central nervous system and plays a role in cerebral damage after hypoxia–ischemia. Excessive NHE activation during hypoxia–ischemia leads to intracellular Na+ overload, which subsequently promotes Ca2+ entry via reversal of the Na+/Ca2+ exchanger. Increased cytosolic Ca2+ then triggers the neurotoxic cascade. Activation of NHE also leads to rapid normalization of pHi and an alkaline shift in pHi. This rapid recovery of brain intracellular pH has been termed pH paradox as, rather than causing cells to recover, this rapid return to normal and overshoot to alkaline values is deleterious to cell survival. Brain pHi changes are closely involved in the control of cell death after injury: an alkalosis enhances excitability while a mild acidosis has the opposite effect. We have observed a brain alkalosis in 78 babies with neonatal encephalopathy serially studied using phosphorus-31 magnetic resonance spectroscopy during the first year after birth (151 studies throughout the year including 56 studies of 50 infants during the first 2 weeks after birth). An alkaline brain pHi was associated with severely impaired outcome; the degree of brain alkalosis was related to the severity of brain injury on MRI and brain lactate concentration; and a persistence of an alkaline brain pHi was associated with cerebral atrophy on MRI. Experimental animal models of hypoxia–ischemia show that NHE inhibitors are neuroprotective. Here, we review the published data on brain pHi in neonatal encephalopathy and the experimental studies of NHE inhibition and neuroprotection following hypoxia–ischemia.

10.2. Acid–base regulation during hypercapnia, exercise and anoxia in the armoured catfish, Liposarcus pardalis

During exposure to elevated water CO2 levels (hypercapnia), blood pH and tissue intracellular pH (pHi) in fish are initially reduced due to the rapid development of a respiratory acidosis. Blood pH recovery is associated with acid–base relevant ion transfer at the gills (i.e., Na+/H+ or Cl−/HCO3−), and generally occurs over 24–48 h during which time pHi recovery closely follows pH recovery of the blood. In the armoured catfish, Liposarcus pardalis, hypercapnia induces a large respiratory acidosis, but intracellular pH (pHi) of tissues such as the liver, heart and muscle is tightly regulated despite large reductions in plasma pH. Using an in situ perfused heart preparation, maximum cardiac performance (power output, cardiac output and stroke volume) was maintained at levels of up to 5% CO2 in L. pardalis. Maintenance of heart function under these conditions was associated with preferential pHi regulation. Following exhaustive exercise and 2 h of anoxia, there was little evidence for lactate incorporation into the skull or bony plates, but liver, heart and brain pHi were preferentially regulated despite a large uncompensated plasma acidosis, indicating that the type of acidosis does not alter the pattern of acid–base regulation in this species. This pattern of preferential pHi regulation differs markedly from that of most fish investigated to date; however, it has also been observed in two other facultative air-breathing fishes, Synbranchus marmoratus and Amia calva, and may be associated with the evolution of air-breathing.

Effect of angiotensin II on cerebral edema following cerebral ischemia and reperfusion

Angiotensin II may be a significant contributor to the pathophysiology of ischemic stroke. The major goal of the present study was to clarify the role of angiotensin II in brain ischemia. We investigated the effect of angiotensin II on cerebral ischemic injury following transient forebrain ischemia induced by bilateral occlusion of the common carotid arteries in pentobarbital anesthetized gerbils. Following 60-min cerebral ischemia, we infused angiotensin II ( n=17), phenylephrine ( n=17), or saline ( n=17), and evaluated their effects on blood pressure, cerebral blood flow, survival ratio, cerebral edema, and cerebral energy metabolism. Angiotensin II treatment during post-cerebral ischemic reperfusion worsened specific gravity of cerebral cortex ( p<0.01) in association with a less recovery of brain pHi ( p<0.005). However, phenylephrine treatment, in the same blood pressure manipulation level and duration as angiotensin II treatment, improved the survival rate ( p<0.05) and specific gravity of cerebral cortex ( p<0.005) compared with saline treatment. Therefore, we believe that angiotensin II may have harmful effects in cerebral ischemia.

Brain alkaline intracellular pH after neonatal encephalopathy.

Experimental studies demonstrate an alkaline shift in brain intracellular pH (pH(i)) after hypoxia-ischemia (HI). In infants with neonatal encephalopathy after HI, our aims were to assess (1) brain pH(i) during the first 2 weeks after birth in infants categorized according to magnetic resonance imaging (MRI) during the first 2 weeks after birth and at more than 3 months of age, and neurodevelopmental outcome at 1 year; (2) the relationship between brain pH(i) and lactate/creatine; and (3) duration of alkaline brain pH(i). Seventy-eight term infants with neonatal encephalopathy were studied using MR techniques. One hundred and fifty-one studies were performed throughout the first year including 56 studies of 50 infants during the first 2 weeks after birth. pH(i) was calculated using phosphorus-31 MR spectroscopy and lactate/creatine was measured using proton MRS. The mean (standard deviation [SD]) brain pH(i) during the first 2 weeks after birth in infants with severely abnormal versus normal MRI was 7.24 (SD, 0.17) versus 7.04 (SD, 0.05; p < 0.001); in infants who subsequently developed cerebral atrophy versus those who did not: 7.23 (SD, 0.17) versus 7.06 (SD, 0.06; p < 0.05); in infants who died or had a severe neurodevelopmental impairment versus normal outcome: 7.28 (SD, 0.15) versus 7.11 (SD, 0.09; p < 0.05). Brain alkalosis was associated with increased brain lactate/creatine (p < 0.001). pH(i) remained more alkaline in the severe outcome group up to 20 weeks after birth (p < 0.05).

Effects of glucose and PaO2 modulation on cortical intracellular acidosis, NADH redox state, and infarction in the ischemic penumbra.

During focal cerebral ischemia, the ischemic penumbra or border-zone regions of moderate cortical blood flow reductions have a heterogeneous development of intracellular cortical acidosis. This experiment tested the hypotheses that (1) this acidosis is secondary to glucose utilization and (2) this intracellular acidosis leads to recruitment of potentially salvageable tissue into infarction. Brain pHi, regional cortical blood flow, and NADH redox state were measured by in vivo fluorescent imaging, and infarct volume was assessed by triphenyltetrazolium chloride histology. Thirty fasted rabbits divided into 6 groups of 5 each were subjected to 4 hours of permanent focal ischemia in the presence of hypoglycemia ( approximately 2.8 mmol/L), moderate hyperglycemia ( approximately 11 mmol/L), and severe hyperglycemia (>28 mmol/L) under either normoxia or moderate hypoxia (PaO2 approximately 50 mm Hg). Preischemic hyperglycemia led to a more pronounced intracellular acidosis and retardation of NADH regeneration than in the hypoglycemia groups under both normoxia and moderate hypoxia in the ischemic penumbra. For example, 4 hours after ischemia, brain pHi in the severe hyperglycemia/normoxia group measured 6.46, compared with 6.84 in the hypoglycemia/normoxia group (P<0.01), and NADH fluorescence measured 173% compared with 114%. Infarct volume in the severe hyperglycemia/normoxia group measured 35.1+/-6.9% of total hemispheric volume, compared with 13.5+/-4.2% in the hypoglycemia/normoxia group (P<0.01). Hyperglycemia significantly worsened both cortical intracellular brain acidosis and mitochondrial function in the ischemic penumbra. This supports the hypothesis that the evolution of acidosis in the ischemic penumbra is related to glucose utilization. Furthermore, the observation that hypoglycemia significantly decreased infarct size supports the postulate that cortical acidosis leads to recruitment of ischemic penumbra into infarction.

Magnesium Deficiency Exacerbates Brain Injury and Stroke Mortality Induced by Alcohol: A 31P-NMR In Vivo Study

Mimicking in rats the reduced level of dietary magnesium (Mg) intake, seen in present-day Western World populations, short-term (4 weeks) restriction of Mg intake (30–35% normal) resulted in a 40% loss in brain intracellular free Mg 2+ ions ([Mg 2+] i) and significant rises in brain intracellular pH (pHi) and phosphocreatine ([PCr]) but no change in [ATP] or [Pi] as measured by 31P-NMR spectroscopy. Such Mg-deficient animals (serum Mg fell 65%), when given ED40 stroke doses of ethanol, demonstrated a 100% stroke mortality. These findings indicate that: 1) moderate, short-term Mg deficiency makes the brain vulnerable to hypoxic-lethal stroke insults induced by alcohol administration, and 2) brain [Mg 2+] i appears to play an important role in finely regulating brain pHi and [PCr].

Nitric oxide synthase inhibition by L-NAME prevents brain acidosis during focal cerebral ischemia in rabbits.

This experiment examined the effects of nitric oxide (NO) synthase inhibition on brain intracellular pH, regional cortical blood flow, and NADH fluorescence before and during 3 h of focal cerebral ischemia using in vivo fluorescence imaging. Thirty fasted rabbits under 1% halothane were divided into four treatment groups receiving N omega-nitro-L-arginine methyl ester (L-NAME) intravenously at 20 min prior to ischemia (0.1, 1, and 10 mg/kg and 1 mg/kg + 5 mg/kg L-arginine) and two control groups (nonischemic and ischemic). In ischemic controls, brain pH(i), declined to 6.73 +/- 0.03 at 30 min and remained acidotic through the remainder of the ischemic period. In the 0.1 mg/kg group, brain pH(i) fell after 30 min of ischemia to 6.76 +/- 0.05 (p < 0.05), but then improved progressively despite occlusion. In the 1 mg/kg group, brain pH(i), remained normal despite middle cerebral artery (MCA) occlusion. In the 10 mg/kg group and in the combined L-NAME + L-arginine group, pH(i) fell after 30 min of ischemia to 6.81 +/- 0.03 (p < 0.05) and remained acidotic. During occlusion, regional cortical blood flow dropped in a dose-dependent manner. After 3 h of ischemia, regional cortical blood flow was 33.9 +/- 10.9 and 25.1 +/- 8.9 ml/100 g/min at doses of 0.1 and 10.0 mg/kg, respectively, L-NAME treatment did not significantly alter the increased NADH fluorescence that accompanied occlusion. This study shows that L-NAME can prevent intracellular brain acidosis during focal cerebral ischemia independent from regional cortical blood flow changes. This experiment suggests that NO is involved in pH(i) regulation during focal cerebral ischemia.

Effects of Intermittent Reperfusion on Brain pHi, rCBF, and NADH During Rabbit Focal Cerebral Ischemia

The use of intermittent reperfusion versus straight occlusion during neurovascular procedures is controversial. This experiment studied the effects of intermittent reperfusion and single occlusion on intracellular brain pH (pHi), regional cerebral or cortical blood flow, and nicotinamide adenine dinucleotide (NADH) fluorescence during temporary focal ischemia. Twenty fasted rabbits under 1.0% halothane anesthesia were divided into four groups: (1) nonischemic controls, (2) 60 minutes of uninterrupted focal ischemia, (3) 2 x 30-minute periods of focal ischemia separated by a 5-minute reperfusion, and (4) 4 x 15-minute periods of focal ischemia separated by three 5-minute reperfusion periods. Focal ischemia was produced by occlusion of both the middle cerebral and ipsilateral anterior cerebral arteries. After the final occlusion, there was a 3-hour reperfusion period in all groups. Regional cerebral and cortical blood flow, brain pHi, and NADH fluorescence were measured with in vivo panoramic fluorescence imaging. During occlusion, regional cerebral and cortical blood flows and NADH fluorescence values were not different among the groups. Brain pHi was significantly lower in the 4 x 15-minute group compared with the 1 x 60-minute group (6.57 +/- 0.02 versus 6.73 +/- 0.06; P < .03) but not significant when compared with the 2 x 30-minute group. During the short reperfusion periods, all parameters returned to normal except for NADH fluorescence levels, which remained elevated. During the postischemic final reperfusion period, there was a mild brain alkalosis of approximately 7.1 in all groups. There were no significant differences in NADH fluorescence among groups during the final reperfusion. Regional cerebral and cortical blood flow returned to near normal values in all groups. This study demonstrates that intermittent reperfusion during temporary focal ischemia has different effects on the intracytoplasmic and the intramitochondrial compartments: worsening of brain cytoplasmic pHi but no significant differences in the oxidation/reduction level of mitochondrial NADH.

Intracellular pH in lizards after hypercapnia

We used the transmembrane distribution of 5,5-[2-14C]dimethyloxazolidine-2,4-dione ([14C]DMO) and 31P magnetic resonance spectroscopy (NMR) to investigate the effects of hypercapnia on intracellular pH (pHi) in brain and skeletal muscle of two lizard species: Anolis equestris and Dipsosaurus dorsalis. In control animals (normocapnic), plasma PCO2 (3.3 +/- 0.1 kPa) and plasma pH (7.52 +/- 0.01) for D. dorsalis were not significantly different from the values for A. equestris (2.8 +/- 0.2 kPa and 7.59 +/- 0.02, respectively). Furthermore 60 min of 5% CO2 increased plasma PCO2 and decreased plasma pH by the same amounts in both species. Brain pHi values determined with the DMO method were not significantly different from values determined with NMR. Control values of brain pHi (DMO, 7.16 +/- 0.01; NMR, 7.11 +/- 0.02) and muscle pHi were significantly higher for D. dorsalis (DMO, 7.15 +/- 0.03) than for A. equestris (DMO, 6.99 +/- 0.03; NMR, 7.02 +/- 0.02 for brain; DMO, 6.97 +/- 0.03 for muscle). In addition, changes in tissue pHi after 60 min of 5% CO2 were significantly different for the two species. In D. dorsalis muscle and brain pHi decreased significantly after hypercapnia, whereas in A. equestris muscle pHi decreased significantly but brain pHi was unchanged. Our findings were independent of the methods used to determine pHi. The smaller change in brain and muscle pHi than in plasma pH for A. equestris is consistent with the view that pHi regulation involves active processes such as transmembrane ion transport.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between intracellular pH, extracellular pH, and ventilation during dilution acidosis.

Previous experiments showed that acute hyperosmolality results in an extracellular acidosis that does not stimulate respiratory compensation (C.E. Kasserra, D. R. Jones, and M. R. Hughes. Respir. Physiol. 85: 383-393, 1991). The data suggested that development of the extracellular dilution acidosis would also result in a concomitant intracellular contraction alkalosis. The effects of acute hyperosmolality and lactacidosis on systemic intracellular pH (pHi) were studied in the conscious Pekin duck in an effort to separate the effects of pHi and extracellular pH (pHe) on ventilatory control. Brain pH was also measured during systemic hyperosmolality to determine the relationship between blood and brain pHi. Hyperosmolality caused a concurrent extracellular acidosis and intracellular alkalosis in pectoral muscle, whereas lactic acid infusion decreased both pHe and pHi. Ventilation was stimulated only during lactacidosis and did not change during hyperosmolality. Brain pHi did not show a consistent significant increase in response to systemic hyperosmolality over 1 h but did show a trend toward an alkalosis. Measurement of high-energy phosphate metabolites (phosphocreatine, ATP, and Pi) indicated an increase of metabolic rate during hyperosmolality. With the assumption that similar pHi changes were occurring in chemoreceptive cells, the results suggest that ventilation was responding to pHi changes and that much of the depressive response to acute hyperosmotic disturbance was peripherally generated.

Acidic foci within the ischemic penumbra of the New Zealand white rabbit.

In vivo panoramic imaging of reduced nicotinamide adenine dinucleotide (NADH), intracellular brain pH (pHi), and cortical blood flow was used to characterize the ischemic penumbra during focal ischemia. During global ischemia, hypoxia, and status epilepticus, the development of cortical acidic foci has been observed. The hypothesis tested was that during focal ischemia, acidic foci develop, which may lead to recruitment of the ischemic penumbra into infarction. Five fasted New Zealand White rabbits underwent middle cerebral artery (MCA) occlusion under 1.5% halothane anesthesia through a retro-orbital approach, and five animals served as controls. Brain pHi and cerebral blood flow were measured with in vivo umbelliferone fluorescence. Baseline brain pHi was 6.98 +/- 0.05, whereas cortical blood flow and NADH fluorescence measured 52.2 +/- 8.7 mL/100 g per minute and 35.5 +/- 3.7 gray-scale units, respectively. Fifteen minutes after MCA occlusion, overall brain pHi and cortical blood flow of the ischemic penumbra measured 6.61 +/- 0.06 and 31.9 +/- 9.2 mL/100 g per minute. Over 3 hours there was normalization of pHi in the majority of the penumbra due to increases in cortical blood flow. Within the ischemic penumbra acidic foci developed with an initial pHi of 6.35 +/- 0.09 and cortical blood flow of 18.0 +/- 5.7 mL/100 g per minute. These foci remained acidic with increased NADH fluorescence despite being surrounded by cortex that was recovering from ischemia. On light microscopy, these acidic foci had a mixed pattern of neuronal injury. Within the ischemic penumbra, acidic foci develop that do not follow a vascular distribution and have microscopic evidence of ischemic neuronal injury. This suggests that there is a cortical selective vulnerability regarding pHi regulation and these acidic foci may lead to recruitment of the ischemic penumbra into infarction.

Effect of superhypercapnia on cortical pHi and cortical blood flow

In vivo panoramic imaging of regional cortical blood flow and intracellular brain pH with umbelliferone fluorescence was performed in fasted New Zealand White rabbits (n = 30) subjected to constant or stepwise increases in arterial PCO2 (PaCO2) up to 160 mmHg under 1.0% halothane anesthesia. At a PaCO2 of 40 mmHg, baseline brain pHi measured 7.03 +/- 0.04, while regional cortical blood flow was 47.0 +/- 4.3 ml.100 g-1.min-1. With an immediate but constant exposure to a PaCO2 of 60, 80, or 120 mmHg, at 15 min pHi fell to 6.94 +/- 0.03, 6.89 +/- 0.03, and 6.90 +/- 0.03 and recovered over 20, 80, and 120 min, respectively. At 160 mmHg PaCO2, pHi fell to 6.68 +/- 0.04 and remained acidotic. With stepwise increases in PaCO2 up to 80 mmHg, brain pHi decreased from 7.01 +/- 0.02 to 6.99 +/- 0.02, not significantly different despite a severe systemic acidosis of 7.022 +/- 0.034. At a PaCO2 of 120 and 160 mmHg, pHi dropped to 6.97 +/- 0.02 and 6.93 +/- 0.02, respectively. The difference in brain pHi between an immediate and stepwise exposure to a PaCO2 of 160 mmHg was significant (P < 0.005). These results demonstrate that brain pHi is tightly regulated across the cortical surface and resistant to profound changes in extracellular pH. The difference in pHi between the constant and incremental increases in PaCO2 suggests that there is upregulation of pH homeostatic mechanisms.

Brain pHi, cerebral blood flow, and NADH fluorescence during severe incomplete global ischemia in rabbits.

The aim of this experiment was to study the serial changes in brain intracellular pH, cerebral blood flow, and the oxidation/reduction level of intramitochondrial nicotinamide adenine dinucleotide fluorescence across the cortical surface during severe incomplete global ischemia. Reduced nicotinamide adenine dinucleotide fluorescence and brain intracellular pH using the pH-sensitive indicator umbelliferone were measured with in vivo panoramic fluorescence imaging of the cortical surface. Cerebral blood flow was measured with the clearance of both umbelliferone and xenon-133. Fifteen minutes of severe incomplete global ischemia was produced by temporary occlusion of the innominate, left carotid, and subclavian arteries in five fasted New Zealand White rabbits. Baseline brain intracellular pH was homogeneous over the exposed cortex, measuring 7.00 +/- 0.02, while cerebral blood flow was 48.0 +/- 2.6 ml/100 g/min. During 15 minutes of ischemia, cerebral blood flow measured 6.3 +/- 1.8 ml/100 g/min and brain pH declined to 6.61 +/- 0.02 (p < 0.005); in addition, there were acidotic foci with pH measuring 6.40 +/- 0.10. During reperfusion, there was an initial normalization of brain intracellular pH without an alkaline shift followed by a recurrent cortical acidosis of pH 6.88 +/- 0.06. There was a heterogeneous pattern of fluorescence that increased significantly following 60 minutes of reperfusion, coinciding with a postischemic hypoperfusion. The hypoperfusion was a uniform reduction in cerebral blood flow over the brain's surface, with reductions of 42.5% and 44.2% at 30 and 45 minutes, respectively. During incomplete global ischemia there is a heterogeneous pattern of brain intracellular pH and reduced nicotinamide adenine dinucleotide changes that do not correlate with changes in cortical blood flow. The acidotic foci that were approximately 0.2 pH units more acidotic than the surrounding cortex may be the result of continued glucose delivery under anaerobic conditions. The degree of reduced nicotinamide adenine dinucleotide fluorescence suggests that the cortex is most vulnerable to metabolic failure after 60 minutes of reperfusion following severe incomplete global ischemia. The heterogeneous pattern of brain intracellular pH and reduced nicotinamide adenine dinucleotide changes suggest that there may be a selective vulnerability of cortical tissue to an ischemic challenge.

Effect of hypoglycemia on changes of brain lactic acid and intracellular pH produced by ischemia.

Previous investigators have attributed the fall of brain intracellular pH (pHi) produced by ischemia to accumulation of lactic acid. The goal of the present experiments was to examine the hypothesis that the acidosis produced by cerebral ischemia is due to accumulation of lactic acid. The present experiments inhibited lactic acid production by lowering glucose availability using insulin-induced hypoglycemia. The adverse effects of hypoglycemia were prevented by the prior elevation of beta-hydroxybutyric acid and acetoacetic acid induced by a high lipid diet. Brain pHi and lactic acid were measured by 31P and 1H NMR. The results showed that insulin-induced hypoglycemia markedly inhibits production of lactic acid, but has no effect on brain pHi during ischemia. These findings suggest that, at least under some conditions, the acidosis produced by cerebral ischemia is not due to accumulation of lactic acid.

Hypocapnic-hypoglycemic interactions on cerebral high-energy phosphates and pH in dogs.

With a level of hypoglycemia (1-1.5 mM) that does not alter cerebral O2 uptake and glucose uptake in dogs, induction of hypocapnia may cause severe electroencephalographic (EEG) abnormalities. The aim of this study was to determine the effect of hypoglycemia (blood glucose = 1.1 +/- 0.1 mM) and hypocapnia (arterial PCO2 = 15 +/- 1 mmHg) on cerebral ATP, phosphocreatine, and intracellular pH (pHi; 31P magnetic resonance spectroscopy), cerebral blood flow (CBF; radiolabeled microspheres), global O2 uptake, and glucose uptake in anesthetized dogs. Neither hypoglycemia nor hypocapnia alone altered brain high-energy phosphates, pHi, O2 or glucose uptake or caused major EEG abnormalities. Hypocapnia alone decreased CBF to 62 +/- 4% of control. The combination of hypoglycemia and hypocapnia did not decrease CBF (85 +/- 6% of control), and O2 and glucose uptake were unchanged. During hypocapnic hypoglycemia, isoelectric EEG was seen in 40% of animals, ATP and phosphocreatine decreased to 38 +/- 12 and 43 +/- 12% of control, respectively, while pHi increased from 7.13 +/- 0.05 to 7.43 +/- 0.09. The increase in pHi was related reciprocally to the decrease in venous PCO2, indicating little change in intracellular bicarbonate concentration ([HCO3-]i). With normoglycemic hypocapnia, in contrast, estimated [HCO3-]i decreased 57 +/- 1%. These data suggest that active regulation of pHi during normoglycemic hypocapnia is impaired during hypoglycemic hypocapnia associated with decreased ATP.

Focal cortical distribution of blood flow and brain pHi determined by in vivo fluorescent imaging.

A technique was developed for panoramic imaging of intracellular focal brain pH (pHi) and focal cortical blood flow to measure the pattern of changes in these parameters during arterial PCO2 (PaCO2) and arterial PO2 (PaO2) alterations. Eleven overnight-fasted New Zealand White rabbits were operated and studied under 2.0 and 1.0% halothane anesthesia, respectively. The exposed cortex was imaged by a 512 x 512 pixel image intensifier system with a resolution of 10 microns2/pixel. Focal brain pHi and focal cortical blood flow were measured by umbelliferone fluorescence. The mean focal cortical blood flow was 49 +/- 4.5 ml.100 g-1.min-1, and focal brain pHi was 7.05 +/- 0.02 during normocapnia. At a stable PaCO2, focal cortical blood flow and focal brain pHi within 10 microns of surface-conducting vessels were 2-50% greater and 0.02 pH units lower than that observed in the border-zone regions between vessels. Focal cortical blood flow demonstrated a significant heterogeneity of 51% among various cortical regions, despite a stable PaCO2. Alternatively, focal brain pHi was homogeneous, varying no more than 0.05 pH units. These results demonstrate the dependence of focal cortical blood flow on topical locations from surface conducting vessels and the tight regulation of brain pHi.

Brain Bioenergetics during Cardiopulmonary Resuscitation in Dogs

Cardiac arrest causes a rapid loss of cerebral adenosine triphosphate [corrected] (ATP) and a decrease in cerebral intracellular pH (pHi). Depending on the efficacy of cardiopulmonary resuscitation (CPR), cerebral blood flow levels (CBF) ranging from near zero to near normal have been reported experimentally. Using 31P magnetic resonance spectroscopy, the authors tested whether experimental CPR with normal levels of cerebral blood flow can rapidly restore cerebral ATP and pHi despite the progressive systemic acidemia associated with CPR. After 6 min of ventricular fibrillation in six dogs anesthetized with fentanyl and pentobarbital, ATP was reduced to undetectable concentrations and pHi decreased from 7.11 +/- 0.02 to 6.28 +/- 0.09 (+/- SE) as measured by 31P magnetic resonance spectroscopy. Application of cyclic chest compression by an inflatable vest placed around the thorax and infusion of epinephrine (40 micrograms/kg bolus plus 8 micrograms/kg/min, intravenously) maintained cerebral perfusion pressure greater than 70 mmHg for 50 min with the dog remaining in the magnet. Prearrest cerebral blood flows were generated. Cerebral pHi recovered to 7.03 +/- 0.03 by 35 min of CPR, whereas arterial pH decreased from 7.41 +/- 0.4 to 7.08 +/- 0.04 and cerebral venous pH decreased from 7.29 +/- 0.03 to 7.01 +/- 0.04. Cerebral ATP levels recovered to 86 +/- 7% (+/- SE) of prearrest concentration by 6 min of CPR. There was no further recovery of ATP, which remained significantly less than control. Therefore, in contrast to hyperemic reperfusion with spontaneous circulation and full ATP recovery, experimental CPR may not be able to restore ATP completely after 6 min of global ischemia despite restoration of CBF and brain pHi to prearrest levels.

Control of ventilation in the hypercapnic skate Raja ocellata: II. Cerebrospinal fluid and intracellular pH in the brain and other tissues

This study examined the possible role(s) of central-base stimuli in the increase in ventilation induced by hypercapnia in the skate, a response that is not due to an O 2 signal (Graham et al., Respir. Physiol., 1990, 80: 251–270). Skate were sampled for cerebrospinal fluid (CSF) acid-base status, intracellular pH of the brain ( 14C-DMO method), and pHi in other tissues throughout 24 h of exposure to Pi CO 2 = 7.5 Torr . CSF P CO 2 rapidly equilibrated with the elevated Pa CO 2 . Despite the much lower non-HCO 3 −1 buffer capacity in the CSF, CSF pH was not depressed to the same extent as blood pHa. CSF pH was also regulated more rapidly, returning to control levels by 8–10 h, whereas pHa remained significantly depressed at 24 h. Similarly, the pHis of the weakly buffered brain and heart ventricle were initially compensated more rapidly than those of more strongly buffered white muscle and red blood cells. However, brain pHi adjustment slowed markedly after 4 h and stabilized at only 70% compensation by 20–24 h, suggesting that brain intracellular acidosis may play a role in the long-term increase in ventilation. CSF and brain were the only compartments which did not exhibit an apparent compounding metabolic acidosis during the initial stages of hypercapnic exposure. While these results illustrate the primacy of central acid-base regulation, they do not support a role for CSF pH in the long-term elevation of ventilation in response to hypercapnia. Depressions in pHa and brain phi appear the two mosly likely candidates for proximate stimuli.