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31P magnetic resonance spectroscopy demonstrates expansion of the extracellular space in the skeletal muscle of starved rats.

Starvation significantly alters the distribution of body water. To study the effects of starvation on cellular energetics and water distribution in skeletal muscle, a novel 31P magnetic resonance technique (31P MRS) was developed to measure water compartments. After 31P MRS-visible water space markers which distribute in total body water (dimethyl methylphosphonate, DMMP) and extracellular water (phenylphosphonate, PPA) were infused intravenously, 31P MRS spectra were obtained from the gastrocnemius muscle of male virus-free Wistar rats at baseline and after starvation or ad libitum feeding for 4 days. Muscle water spaces were also measured using the chloride method and Nernst's equation. Muscle water contents as determined by drying were equivalent in the two groups. In vivo measurements of changes in DMMP relative to all of the MRS visible phosphates also demonstrated that the total water space was similar in control and starved rats. However, starvation significantly increased the ratio of PPA/DMMP (0.67 +/- 0.05 vs 0.87 +/- 0.04, Control vs Starvation; P < 0.001), and therefore the ratio of extracellular water to total water in the gastrocnemius. Furthermore, because muscle water contents were comparable between the groups, this expansion of the extracellular space was accompanied by contraction of the intracellular compartment in starved animals. Equivalent changes were detected in vitro using the chloride method. Lastly, phosphocreatine/ATP ratios, which measured changes in high-energy phosphate stores, decreased after starvation (4.09 +/- 0.06 vs 3.61 +/- 0.06; P < 0.001) and were inversely related to changes in PPA/DMMP (r = -0.61; P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

Increase of catecholamine content in the extracellular space of the rat's brain cortex during spreading depression wave as determined by voltammetry.

The effect of chemically initiated (KCl) spreading cortical depression on catecholamine content in the extracellular cortical space was examined using voltammetry technique in chloralhydrate-anaesthetized rats. Correlation between alterations in catechol-oxidative current (CA.OC) and the time-course of the slow potential (SP) change (one of the chief features of the spreading depression) showed that spreading depression wave was accompanied by the significant increase in CA.OC content (up to 158 +/- 43%, mean +/- S.D., P < 0.001). The rise of the negative SP preceded significant CA.OC increase by 20 +/- 8 s. This fact provides the evidence that catecholamine overflow cannot participate in triggering spreading depression wave; nevertheless it can be an important link of the spreading depression mechanism.

Animals↗

Theophylline concentration in the extracellular space of the rat brain: measurement by microdialysis and relation to behaviour.

The free extracellular concentration of theophylline in the brain was estimated from microdialysis samples. Two different methods were used to estimate extracellular concentrations by microdialysis, the perfusion rate method and the difference method. Theophylline 20 mg/kg (s.c.) gave a sufficiently stable level of theophylline in the brain 60 min after injection and lasting over the observation period to allow application of the two methods in vivo. The relation between dose and dialysate concentration was linear. It was found that doses of 20-24 mg/kg theophylline corresponded to a free extracellular concentration of 60-90 microM. The behaviour of theophylline-treated rats was assessed in parallel experiments by means of a holeboard apparatus. Behavioural activation was observed in the dose-range 3-30 mg/kg. It is concluded that behavioural effects of theophylline can be induced at a concentration well below that required to inhibit phosphodiesterase but within the range in which adenosine receptor blockade may be observed, suggesting that the latter mechanism is responsible for the behavioural effects of theophylline.

Animals↗

K+ changes in the extracellular space of the spinal cord and their physiological role.

K+ accumulates in the intercellular space as a result of neuronal activity. The changes in extracellular K+ concentration, delta[K]e (estimated by K+-selective microelectrodes), depends on neuronal activity, on the density of discharging neurones and the removal of the accumulated K+ by diffusion, active transport and current flow through cells. In the mammalian as well as the amphibian spinal cord a single volley in a peripheral nerve increases [K]e by 0.2-0.5 mmol . 1-1, while tetanic stimulation (100 Hz) by 7-8 m-mol . 1-1, with a maximum in the lower dorsal horn. Increased [K]e was also found in lumbar segments when the somatosensory cortex of the cat and medulla of the frog were stimulated. In the frog spinal cord, the tactile stimulation of the hindlimb evoked delta[K]e by about 0.1 mumol . 1-1, nociceptive stimulation by 0.2-1.0 mmol . 1-1. Spontaneous delta[K]e and dorsal root potentials (DRPs) were observed at various intervals after stimulation, associated with the decay phase of delta[K]e. It was shown that primary afferent depolarization (PAD) consists of two components: the 'early' component (mediated by GABA and depressed by picrotoxin or bicuculline) and the 'late' K+ component (potentiated by picrotoxin and bicuculline). Even when increased [K]e produces PAD, this does not mean that it also results in presynaptic inhibition. It was found that the delta[K]e produced depolarization of motoneurones and neuroglia and there is every reason to believe that this also applies to the interneurones. Evidence is available that an increase of [K]e up to 6 mmol . 1-1 facilitates impulse transmission in the spinal cord while higher levels result in its inhibition.

Afferent Pathways↗

Cat heart muscle in vitro. III. The extracellular space.

The "osmotic gradient" method, an intracellular microelectrode technique for determining whether an uncharged, water-soluble molecule enters cells or remains extracellular, is described. Using this method, a series of carbohydrates of graded molecular size were examined. In cat papillary muscles mannitol, molecular radius 4.0 A, remained extracellular while arabinose, molecular radius 3.5 A entered the cells. Measurement of the simultaneous uptake of H(3)-mannitol and C(14)-inulin showed that mannitol equilibrates with 40 per cent of total water in 1 hour, after which the mannitol space does not further increase. By contrast, inulin, molecular radius approximately 15 A, equilibrates with 24 per cent of total water in 1 hour; thereafter the inulin space continues to increase very slowly. The intracellular K concentrations are significantly higher and the intracellular Na and Cl concentrations significantly lower when mannitol rather than inulin is used to measure the extracellular space. The intracellular Cl concentration determined with Cl(36) or Br(82) is significantly higher than that calculated from the membrane potential assuming a passive Cl distribution. In addition, it is shown that choline enters and is probably metabolized by the cells of papillary muscle.

Animals↗