Biochemical and ultrastructural analyses of IgLON cell adhesion molecules, Kilon and OBCAM in the rat brain.

Kilon (kindred of IgLON) and opioid-binding cell adhesion molecule belong to the IgLON subgroup of immunoglobulin superfamily together with the limbic system-associated membrane protein and neurotrimin. In the present study, we have analyzed biochemical and ultrastructural characterization of Kilon and opioid-binding cell adhesion molecule such as regional and developmental expression patterns, light and electron microscopic localization, and intermolecular interactions. Western blotting revealed a widespread distribution pattern of Kilon with high expression levels in the olfactory bulb, cerebral cortex, diencephalon, hippocampus, and cerebellum and low expression levels in the medulla oblongata and spinal cord. In contrast, opioid-binding cell adhesion molecule showed a regionally restricted expression pattern with high levels only in the cerebral cortex and hippocampus. Expression of Kilon and opioid-binding cell adhesion molecule was increased gradually during postnatal development and maintained until adulthood. Light microscopic immunohistochemistry demonstrated that the localization of opioid-binding cell adhesion molecule and Kilon coincided well with that of vesicle-associated membrane protein 2, a synaptic marker protein, in the cerebral cortex and hippocampus of adult brain. In the cerebellum, Kilon-immunoreactive puncta were observed to colocalize well with that of vesicle-associated membrane protein 2, while opioid-binding cell adhesion molecule immunoreactivity was observed only at part of synaptic glomeruli in the granular layer and rare in the molecular layer. Electron microscopic analysis revealed that Kilon and opioid-binding cell adhesion molecule immunoreactivity was observed mainly at postsynaptic sites of dendritic and somatic synapses in adult cerebral cortex and hippocampus. Only trace levels of Kilon and opioid-binding cell adhesion molecule were detected in the soluble fraction of a cortical homogenate, although a substantial amount of F3 was present in the soluble fraction. A binding analysis using a cross-linker and the immunoprecipitation technique demonstrated that Kilon and opioid-binding cell adhesion molecule interacted heterophilically and homophilically. These findings show that Kilon and opioid-binding cell adhesion molecule are clearly distinguishable from each other in regional expression and localization, and binding patterns. These differences possibly represent diverse functions of each IgLON molecule.

Changes in the localization of NAP-22, a calmodulin binding membrane protein, during the development of neuronal polarity.

NAP-22, a neuronal tissue-enriched acidic membrane protein, is a Ca(2+)-dependent calmodulin binding protein and has similar biochemical characteristics to GAP-43 (neuromodulin). Recent biochemical studies have demonstrated that NAP-22 localizes in the membrane raft domain with a cholesterol-dependent manner. Since the raft domain is assumed to be important to establish and/or to maintain the cell polarity, we have investigated the changes in the localization of NAP-22 during the development of the neuronal polarity in vitro and in vivo, using cultured hippocampal neurons and developing cerebellum neurons, respectively. Cultured hippocampal neurons initially extended several short processes, and at this stage NAP-22 was distributed more or less evenly among them. During the maturation of neuronal cells, NAP-22 was sorted preferentially into the axon. Throughout the developmental stages of hippocampal neurons, the localization change of NAP-22 was quite similar to that of tau, an axonal marker protein, but not to that of microtubule-associated protein-2 (MAP-2), a dendritic marker protein. Further confocal microscopic observation demonstrated the colocalization of NAP-22 and either tau or vesicle-associated protein-2 (VAMP-2). A comparison of the time course of the axonal localization of NAP-22 and GAP-43 showed that NAP-22 localization was much later than that of GAP-43. The correlation between the expression of NAP-22 and synaptogenesis in the cerebellar granular layer, particularly in the synaptic glomeruli, was also investigated. There existed many VAMP-2 positive synapses but no NAP-22 positive ones in 1-week-old cerebellum. On sections of 2-week-old cerebellum, accumulation of NAP-22 to the synaptic glomeruli was clearly observed and this accumulation became clearer during the maturation of the synaptic structure. The present results suggest the possibility that NAP-22 plays an important role in the maturation and/or the maintenance of synapses rather than in the process of the axonal outgrowth, by controlling cholesterol-dependent membrane dynamics.

Expression of the IgLON cell adhesion molecules Kilon and OBCAM in hypothalamic magnocellular neurons.

The vasopressin (AVP) and oxytocin (OXT) magnocellular neurons in the hypothalamic supraoptic (SON) and paraventricular nuclei (PVN) display reversible structural plasticity of neurons and glial cells under different conditions of neuropeptide secretion. In the present study, we investigated the expression of two immunoglobulin superfamily (IgSF) proteins, Kilon and OBCAM, in the magnocellular neurons by using monoclonal antibodies. Anti-Kilon antibody reacted specifically with the bacterially expressed recombinant Kilon but not with the recombinant OBCAM, and similarly anti-OBCAM antibody specifically recognized the recombinant OBCAM. Western blotting analysis revealed the specific expression of Kilon and OBCAM in the SON homogenates. Although Kilon and OBCAM of the SON homogenates were present as the insoluble form, most Kilon was present in the Triton-insoluble fraction, and OBCAM was localized mainly in the Triton-soluble fraction. Immunocytochemistry revealed Kilon and OBCAM immunoreactivity in the magnocellular neurons of the SON and PVN of the rat hypothalamus compared with outside of the SON and PVN in the hypothalamus. The double-labeling study with confocal microscopy further demonstrated that Kilon immunoreactivity was observed mainly in the dendrites of AVP-secreting neurons and also occasionally OXT-secreting neurons. However, OBCAM immunoreactivity was exclusively seen in the dendrites of AVP-secreting magnocellular neurons. Chronic physiological stimulation by 2% NaCl had no effect on the expression levels of either IgLON protein in the SON. Our study thus demonstrated specific expression of Kilon and OBCAM in the hypothalamic magnocellular neurons, particularly in dendrites, suggesting that they confer on magnocellular neurons the ability to rearrange dendritic connectivity.

Identification of gelsolin as an actin regulatory component in a triton insoluble low density fraction (raft) of newborn bovine brain.

A membrane microdomain enriched in cholesterol and glycosphingolipids, or so called 'raft' region, was found to contain many signal transducing proteins such as GPI-anchored cell adhesion molecules, trimeric G proteins, and protein tyrosine kinases. In previous studies, we showed that the raft region obtained from rat brain contains two cytoskeletal proteins, tubulin and actin, as the major components in addition to these signal transducing proteins. In this study, to know the biochemical mechanisms regulating the cytoskeletal organization in this region, actin regulatory activities in raft were surveyed. We found the presence of a Ca(2+)-dependent actin nucleation promoting activity in raft. The solubilization and column fractionation of this activity combined with western blotting and immunoprecipitation showed that gelsolin is one of the actin regulatory proteins in raft.

Cholesterol-dependent localization of NAP-22 on a neuronal membrane microdomain (raft).

A membrane microdomain called raft has been under extensive study since the assembly of various signal-transducing molecules into this region has been envisaged. This domain is isolated as a low buoyant membrane fraction after the extraction with a nonionic detergent such as Triton X-100. The characteristic low density of this fraction is ascribed to the enrichment of several lipids including cholesterol. To clear the molecular mechanism of raft formation, several extraction methods were applied to solubilize raft components. Cholesterol extraction using methyl-beta-cyclodextrin was found to be effective to solubilize NAP-22, a neuron-enriched Ca(2+)-dependent calmodulin-binding protein as well as one of the main protein components of brain raft. Purified NAP-22 bound to the liposomes that were made from phosphatidylcholine and cholesterol. This binding was dependent on the amount of cholesterol in liposomes. Calmodulin inhibited this binding in a dose-dependent manner. These results suggest that the presence of a calcium-dependent regulatory mechanism works on the assembly of raft within the neuron.

Nerve growth factor inducer, 4-methyl catechol, potentiates central sensitization associated with acceleration of spinal glutamate release after mustard oil paw injection in rats.

1. In rats, injection of mustard oil (MO) into the paw caused a gradual increase in flinching of the injected paw and this algogenic behavior corresponded with an increase in the CSF-Glu level. 2. The nerve growth factor (NGF) inducer, 4-methyl catechol (4MC), enhanced the frequency of flinching and this effect was dose dependent. In addition, spinal CSF-Glu release was significantly above baseline 10 min after MO injection. In contrast, morphine (MOR) pretreatment completely blocked this behavioral and neurohumoral effect. 3. Anti-NGF paw injection attenuated the algogenic behavior and spinal Glu release otherwise observed after 4MC treatment. 4. The results demonstrated that MO-induced hyperalgesia is associated with increased CSF-Glu release and that this effect is potentiated by a NGF inducer. These data also suggest a possible involvement of NGF in the development of central sensitization after acute peripheral nociceptive stimulation.

Characterization of a novel rat brain glycosylphosphatidylinositol-anchored protein (Kilon), a member of the IgLON cell adhesion molecule family.

In the central nervous system, many cell adhesion molecules are known to participate in the establishment and remodeling of the neural circuit. Some of the cell adhesion molecules are known to be anchored to the membrane by the glycosylphosphatidylinositol (GPI) inserted to their C termini, and many GPI-anchored proteins are known to be localized in a Triton-insoluble membrane fraction of low density or so-called "raft." In this study, we surveyed the GPI-anchored proteins in the Triton-insoluble low density fraction from 2-week-old rat brain by solubilization with phosphatidylinositol-specific phospholipase C. By Western blotting and partial peptide sequencing after the deglycosylation with peptide N-glycosidase F, the presence of Thy-1, F3/contactin, and T-cadherin was shown. In addition, one of the major proteins, having an apparent molecular mass of 36 kDa after the peptide N-glycosidase F digestion, was found to be a novel protein. The result of cDNA cloning showed that the protein is an immunoglobulin superfamily member with three C2 domains and has six putative glycosylation sites. Since this protein shows high sequence similarity to IgLON family members including LAMP, OBCAM, neurotrimin, CEPU-1, AvGP50, and GP55, we termed this protein Kilon (a kindred of IgLON). Kilon-specific monoclonal antibodies were produced, and Western blotting analysis showed that expression of Kilon is restricted to brain, and Kilon has an apparent molecular mass of 46 kDa in SDS-polyacrylamide gel electrophoresis in its expressed form. In brain, the expression of Kilon is already detected in E16 stage, and its level gradually increases during development. Kilon immunostaining was observed in the cerebral cortex and hippocampus, in which the strongly stained puncta were observed on dendrites and soma of pyramidal neurons.

[Efficacy of MR angiographic original images on surgery for posterior communicating artery aneurysms].

This study evaluated the usefulness of axial source images of magnetic resonance angiography (MRA) on preoperative depiction of surgical topography around posterior communicating artery aneurysms. Twenty patients with posterior communicating artery aneurysms, two ruptured and eighteen unruptured, underwent conventional angiography as well as axial source and projection images obtained by three-dimensional time-of-flight MRA techniques. By comparing the topography based on these angiograms to that confirmed during surgery, we evaluated useful information specific to the source images of MRA. Source images of MRA visualized the posterior communicating artery and the anterior choroidal artery in eighteen cases (90%) and five cases (25%), respectively. The posterior communicating artery was recognized at a higher rate by source images of MRA than by conventional angiography (65%), while the anterior choroidal artery was recognized at a lower rate than by combined angiography (75%). We realized some specific information to the source images of MRA including the topographical relations between the aneurysmal neck and the orifice of the posterior communicating artery, the relations between the aneurysmal dome and the oculomotor nerve and the aneurysmal dome buried into the temporal lobe. The information suggested a satisfactory direction of safe aneurysmal clipping so as not to occlude the posterior communicating artery. It was concluded that the source images of MRA provided additional useful information on surgical topography in 60% of the cases involving posterior communicating artery aneurysms. Although not essential in every case, the information would be beneficial in cases with the aneurysmal dome suspected to be in the temporal lobe or when the surrounding topography can not be clearly understood by angiography.

Identification of NAP-22 and GAP-43 (neuromodulin) as major protein components in a Triton insoluble low density fraction of rat brain.

NAP-22 is a membrane-localized brain enriched acidic protein having a Ca(2+)-dependent calmodulin binding activity. Further fractionation of the NAP-22 containing membrane showed the localization of NAP-22 in a Triton insoluble fraction of low density. Besides NAP-22, this fraction was found to contain GAP-43 (neuromodulin), trimeric G proteins, and some GPI-anchored proteins such as Thy-1 and N-CAM-120. Presence of some protein tyrosine kinases, such as src and fyn, was also shown.

[Assessment of ventriculoarterial coupling in humans by transesophageal echocardiography and radial artery pressure tracing].

To investigate coupling between the heart and arterial system in patients undergoing elective non-cardiac surgery, we determined both the ventricular elastance and the effective arterial elastance in two groups of subjects: normal group, 68 subjects without heart disease; and cardiac group, 33 subjects with heart disease. Left ventricular end-systolic (Ves) and end-diastolic (Ved) volumes were determined by transesophageal echocardiography. By assuming that left ventricular end-systolic pressure (Pes) is close to mean arterial blood pressure (MAP) and that x-axis intercept (Vo) is zero, the ventricular elastance (E'max) was approximated as MAP/Ves, and the effective arterial elastance (Ea) as MAP/(Ved-Ves). In 222 (74%) of the 299 measurements obtained in normal group, Ea/E'max was nearly 0.5, which is a condition for a maximal mechanical efficiency, while in 61 measurements (20%) Ea was almost equal to E'max (Ea/E'max = 1), which is a condition for maximal stroke work from a given end-diastolic volume. In contrast, in cardiac group, Ea/E'max was nearly 0.5 in 56 (41%) of the 137 measurements, while in 42 measurements (31%) Ea/E'max was nearly 1. In addition, although the value of Ea/E'max over 2, which represents severe heart failure, was not observed in normal group, Ea/E'max was over 2 in 10 measurements (7%) in cardiac group. Thus, the present results suggest that, as reported previously in awake patients, ventriculoarterial coupling is set toward higher left ventricular work efficiency in surgical patients without heart disease, whereas in patients with heart disease, ventricular and arterial properties are so matched as to maximize stroke work at the expense of the work efficiency.

[Assessment of left ventricular contractility (Emax) and arterial load (Ea) in humans by transesophageal echocardiography and radial artery pressure tracing].

We determined both the slope of the left ventricular end-systolic pressure-volume relation (Emax), which is a measure of contractility independent of loading conditions, and the slope of the arterial end-systolic pressure-stroke volume relation (Ea), which is a measure of arterial load independent of ventricular function, in 10 patients undergoing elective noncardiac surgery. Left ventricular end-systolic volume (Ves) was measured by transesophageal echocardiography and instantaneous left ventricular end-systolic pressure (Pes) was estimated from the dicrotic notch pressure in the radial artery. Emax was calculated during afterload reduction (nicardipine 30 micrograms.kg-1 iv), and the correlation of Emax to either Pes/Ves ratio or MAP (mean arterial blood pressure)/Ves ratio was accomplished in order to investigate whether these indices were clinically useful measurements of ventricular function or not. Ea was also calculated from the data obtained before and 2-3 min after nicardipine iv. The averaged Emax and x-axis intercept (Vo) were 3.11 mmHg.ml-1 and -3.8 ml, respectively. The correlation coefficient obtained between Emax and Pes/Ves was 0.96, and that obtained between Emax and MAP/Ves was 0.97. Ea decreased significantly (P < 0.05) following intravenous nicardipine, demonstrating a decreased arterial load. The direction of changes in Ea was similar to that reported previously in systemic vascular resistance. From these results, we conclude that measurement of Emax (or Pes/Ves, MAP/Ves) and Ea using transesophageal echocardiography and radial artery pressure tracing is feasible and these are a useful tool to estimate left ventricular performance and arterial load during surgery.

[Effects of isosorbide dinitrate on regional cerebral blood flow and intracranial pressure in cats].

The effects of isosorbide dinitrate (ISDN) on regional cerebral blood flow (rCBF) and intracranial pressure (ICP) were examined in cats. A low dose of ISDN (2.5 micrograms/kg/min) infusion did not show any changes in cerebral hemodynamics. During high dose of ISDN (5.0 micrograms/kg/min) or NTP (5.0 micrograms/kg/min) infusion, mean blood pressure (mBP) decreased by 10 to 20% accompanied by decreased cerebral perfusion pressure (CPP: mBP-ICP), however, rCBF or ICP did not change. It is concluded that intravenous administrations of ISDN in a dose of 2.5-5.0 micrograms/kg/min that produce slight decrease in blood pressure did not influence on cerebral hemodynamics.

Cerebral blood flow response to PaCO2 during hypothermic cardiopulmonary bypass in rabbits.

Differences in cerebral blood flow (CBF) between alpha-stat and pH-stat management depend on preserved responsiveness of the cerebral vasculature to changes in arterial carbon dioxide tension (PaCO2). We tested the hypothesis that hypothermia-induced reductions in CBF would decrease the CBF response to changing PaCO2 (delta CBF/delta PaCO2). Anesthetized New Zealand white rabbits were randomly assigned to one of three temperature groups--group 1 (37 degrees C, n = 9); group 2 (31 degrees C, n = 10); or group 3 (25 degrees C, n = 10)--and were cooled using cardiopulmonary bypass. After esophageal temperature equilibration (approximately 40 min), oxygenator gas flows were serially varied to achieve PaCO2 values of 20, 40, and 60 mm Hg (temperature-corrected). All animals were studied at all three PaCO2 levels in random order. At each level of PaCO2, CBF and masseter blood flow were determined using radiolabeled microspheres. There were no significant differences between groups with respect to mean arterial pressure (approximately 80 mmHg), central venous pressure (approximately 4 mmHg), or hematocrit (approximately 22%). Prior normothermic studies have found delta CBF/delta PaCO2 to be proportional to CBF. Nevertheless, in this study, with hypothermia-induced reductions in CBF, delta CBF/delta PaCO2 was not significantly different between temperature groups. Thus, hypothermia either increased the sensitivity of the cerebral vasculature to carbon dioxide and/or increased the effective level of cerebrospinal fluid respiratory acidosis produced by each increment of temperature-corrected PaCO2.(ABSTRACT TRUNCATED AT 250 WORDS)

Differences in cerebral blood flow between alpha-stat and pH-stat management are eliminated during periods of decreased systemic flow and pressure. A study during cardiopulmonary bypass in rabbits.

Prior reports suggest cerebral blood flow (CBF) responses to changing bypass (systemic) flow rates may differ between alpha-stat and pH-stat management. To compare the effect of blood gas management upon CBF responses to changing systemic flow and pressure, 15 New Zealand White rabbits, anesthetized with fentanyl and diazepam, underwent nonpulsatile cardiopulmonary bypass at 25 degrees C. One group of animals (n = 8) was randomized to alpha-stat blood gas management that maintained arterial carbon dioxide tension (PaCO2) approximately 40 mmHg when measured at 37 degrees C. A second group (n = 7) was managed with pH-stat technique, maintaining PaCO2 approximately 40 mmHg when corrected to the animal's actual temperature. Bypass was initiated at a flow rate of 100 ml.kg-1.min-1 and, after approximately 20 min, control hemodynamic and CBF measurements (radioactive microspheres) were made. Thereafter, bypass flow rate was changed in random order at 15-min intervals to 50, 70, and 100 ml.kg-1.min-1. CBF and hemodynamic measurements were repeated at the end of each period of altered bypass flow. Groups differed significantly with respect to both pHa and PaCO2. There were no significant differences between groups with respect to bypass flow rate, mean arterial pressure (MAP), central venous pressure, temperature, hematocrit, arterial oxygen tension (PaCO2), or bypass duration at any measurement point. MAP decreased significantly, from approximately 80 to approximately 65 mmHg with decreasing bypass flow (P = 0.0001). Over the entire range of bypass flows, CBF decreased with decreasing bypass flow (P = 0.001), and the degree of change was equivalent among regions and between groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effect of oncotic pressure on cerebral and extracerebral water content during cardiopulmonary bypass in rabbits.

To study the effect of oncotic pressure on brain water content during cardiopulmonary bypass (CPB), 14 anesthetized New Zealand White rabbits underwent 60 min of nonpulsatile CPB at normothermia. Animals were grouped according to the composition of the circuit priming fluid. Group 1 animals (n = 7) received a priming fluid (6.5% hydroxyethyl starch in 0.72 N NaCl; 323 +/- 13 mOsm/kg [mean +/- SD]) that maintained normal colloid oncotic pressure (COP) during CPB (19.0 +/- 1.5 mmHg). Group 2 animals (n = 7) received a priming fluid (0.9 N NaCl; 324 +/- 23 mOsm/kg) that led to a hypooncotic state (COP = 6.2 +/- 1.2 mmHg). Blood chemistries and hemodynamics were recorded every 15 min during CPB. Animals were given additional priming fluid and sodium bicarbonate during CPB to maintain a circuit flow of 85 ml.kg-1.min-1 and arterial pH greater than 7.35. There were no significant differences between groups 1 and 2 with respect to temperature, central venous pressure, mean arterial pressure, PaO2, PaCO2, plasma sodium concentration, or osmolality at any time during CPB, although osmolality increased in both groups. After 60 min of bypass, animals were killed and organ water contents were determined by wet/dry weight ratios. A separate group of nine similarly prepared and anesthetized animals that did not undergo cannulation or CPB also underwent measurement of plasma chemistries and tissue water contents and served as nonbypass controls (group 3). Brain and kidney water contents were unaffected by oncotic pressure, whereas duodenum and skeletal muscle had significantly greater water content (P = 0.003 and P = 0.008, respectively) after hypooncotic CPB.(ABSTRACT TRUNCATED AT 250 WORDS)

[MR imaging of traumatic cerebellar dysfunction].

Four cases of cerebellar dysfunction following head trauma are presented. Cerebellar signs revealed were those such as dysmetria, dysdiadochokinesis, horizontal nystagmus and ataxia. T2-weighted magnetic resonance (MR) imaging (0.15 tesla, spin-echo method; TR 2000 msec. and TE 100 msec.) revealed focal lesions in these patients, although CT scan failed to demonstrate any changes in the cerebellum. The cerebellar symptoms were maximal immediately after the trauma but improved gradually in two cases. These are compatible with the transient traumatic cerebellar dysfunction postulated by R. C. Cantu in 1969. The pathophysiology of this syndrome, whether it is due to cerebellar concussion or contusion, has not yet been determined. The abnormality of the cerebellum revealed by MR imaging seemed to be contusion rather than concussion. Therefore the authors presume that transient traumatic cerebellar dysfunction is caused by minor cerebellar contusion. In the other two cases, delayed epidural hemorrhage ensued and the symptoms disappeared rapidly after evacuation of the hematoma. In these patients, occurrence of delayed epidural hematoma in the posterior cranial fossa was predicted by MR imaging. The authors regard the lesion as an alarm signal indicating the probable occurrence of infratentorial hematoma.

Analysis of MRI and SPECT in patients with acute head injury.

Traumatic lesions defined by magnetic resonance (MR) imaging were divided into two groups according to findings on computed tomography (CT). This classification reflected difference in the regional cerebral blood flow (rCBF). In the contusional lesions which CT could demonstrate, rCBF varied from hyperperfusion to hypoperfusion, while it was almost always decreased in the lesions which CT could not detect. These results suggest that the former may include a mixture of brain oedema and hyperemia and the latter may imply brain oedema. MR imaging can reveal the minor oedema which CT fails to show in patients with acute head injury.

Correlation between arterial blood pressure and oxygenation in tetralogy of Fallot.

To examine the relationship between arterial blood pressure and oxygenation in patients undergoing complete surgical correction of tetralogy of Fallot, a retrospective study of 16 patients was first performed, looking at the correlation between mean arterial blood pressure (MAP) and arterial oxygen tension (PaO2). The correlation between phenylephrine-induced changes in MAP (delta MAP) and those in PaO2 (delta PaO2) was investigated prospectively in seven patients. In the retrospective study, there was a significant correlation between MAP and PaO2 (n = 66; r = 0.55; P less than 0.01), and most data points with a PaO2 less than 50 mm Hg were associated with a MAP less than 60 mm Hg. In the seven patients who received phenylephrine, 10 micrograms/kg, a significant correlation was found between delta MAP and delta PaO2 (n = 10; r = 0.95; P less than 0.01). These results suggest that in tetralogy of Fallot arterial blood pressure is a determinant of arterial oxygenation, and that the risk of serious hypoxia is significant when MAP is less than 60 mm Hg.

Metabolic activation of intercortical and corticothalamic pathways during enflurane anesthesia in rats.

The purpose of this study was to examine the effects of enflurane on local cerebral glucose utilization (LCGU), and to provide further insight into the mechanism of the epileptogenic properties of enflurane. Twenty-four male Wistar rats were divided into four groups; three groups with intact cortex received 0.5, 2, or 4% enflurane, and one group with unilateral cortex excised received 4% enflurane. LCGU was measured at each anesthetic concentration using the autoradiographic 2-[14C]deoxyglucose method. LCGU in ten of 33 structures examined during 2% enflurane decreased by 19-33%, and LCGU in 22 structures during 4% enflurane decreased by 19-65%, when compared with that during 0.5% enflurane. While LCGU, in most structures, decreased in a dose-related manner, LCGU in the corpus callosum, thalamic ventrobasal complex, and hippocampal CA3 field during 4% enflurane increased by 31-70%, compared with that during 0.5% and/or 2% enflurane. With unilateral cortical excision during 4% enflurane, the increase in LCGU in the ventrobasal complex was obliterated in the excision side, and the increase in the corpus callosum was attenuated. High LCGU in the hippocampal CA3 field and contralateral ventrobasal complex was not affected with cortical excision. These results indicate that intercortical and corticothalamic pathways are metabolically activated during deep enflurane anesthesia, suggesting that the epileptogenic property of enflurane is related to activation of these pathways.