ABSTRACT
We used immunocytochemistry to determine the regional and temporal distribution of Fos protein expression in awake and unrestrained rats after a unilateral stereotaxic microinjection of a cholinergic agonist, carbachol, in the thalamic ventroposterolateral and reticular nuclei, previously shown to cause limbic and generalized convulsive seizures. The microinjection of carbachol elicits behavioral alterations including immobilization, staring, facial and jaw clonus, rearing, and falling, followed by recurrent generalized convulsive seizures, and a pattern of c-fos expression throughout the brain. In addition to the hypothalamic paraventricular and supraoptic nuclei, the initial induction of c-fos expression was observed as early as 15 minutes after the carbachol microinjection, in the piriform and entorhinal cortices, the thalamic paraventricular, the supramammilary, the lateral parabrachial nuclei, and the central gray. From 30 minutes to 2 hours, corresponding to the occurrence of motor expression of limbic and recurrent generalized convulsive seizures, Fos immunoreactivity was seen in a number of functionally related brain regions including the hippocampus, the amygdala, and the anterior thalamic nucleus (limbic system); the thalamus, the basal ganglia, and the cortex (thalamo-striatal-cortical system); and the hypothalamus, the central nucleus of the amygdala, the pons, and the medulla (central autonomic system). On the basis of the present results showing regional and temporal c-fos expression and well known neuroanatomical connections, we have constructed a neural network relating the limbic, thalamo-striatal-cortical, and central autonomic systems. This analysis provides, for the first time, neuronal circuits and pathways relating epilepsy-elicited behavioral expression of convulsive seizures and adaptive homeostatic responses and could serve as a basis for studying central autonomic regulation during epileptic disorders.
Subject(s)
Autonomic Nervous System/physiopathology , Cerebral Cortex/physiopathology , Cholinergic Agonists/toxicity , Corpus Striatum/physiopathology , Epilepsy/physiopathology , Kindling, Neurologic/physiology , Limbic System/physiopathology , Thalamus/physiopathology , Acetylcholine/physiology , Amygdala/physiopathology , Animals , Behavior, Animal , Carbachol/toxicity , Convulsants/toxicity , Epilepsy/chemically induced , Genes, fos , Hippocampus/physiopathology , Hypothalamus/physiopathology , Microinjections , Models, Neurological , Motor Activity , Nerve Net/physiopathology , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/genetics , Proto-Oncogene Proteins c-fos/biosynthesis , Rats , Rats, Wistar , Stereotaxic TechniquesABSTRACT
PURPOSE: By estimating the anatomical distribution of neurons expressing c-fos protein, we sought to establish whether the intrinsic neural systems known to be implicated in the cerebrovascular regulation were activated during the increase in cortical blood flow associated with epileptic seizures. METHODS: A single unilateral microinjection of the cholinergic agonist, carbachol, in the thalamic generalized convulsive seizure area was used in anesthetized rats to elicit recurrent episodes of electrocortical epileptiform activity and an increase in cortical blood flow. Neuronal expression of Fos protein was analyzed to identify activated brain regions. RESULTS: We identified two cortical vasodilatory responses: a sustained cortical vasodilatory response associated with the continuous low-frequency, high-amplitude spiking and a transient cortical vasodilatory response invariably related to the recurrent spike-burst activity. The sustained cortical blood flow began to increase at 55-65 min, remaining significantly (p < 0.05) increased and reaching at the end of the experiment < or =182+/-17% of the prestimulated control. The electrocortical epileptic activity and the cerebral cortical vasodilation were associated with a marked increase in Fos immunoreactivity in the entorhinal and piriform cortices, the dentate gyrus, the hippocampus, and the amygdala. Fos-positive neurons also were found in specific thalamic nuclei, the cerebral cortex, the caudate-putamen, the hypothalamus, the pontine parabrachial nuclei, the dorsal raphe, and the rostral ventrolateral medulla. CONCLUSIONS: These results provide evidence that convulsive seizures elicited by cholinergic stimulation of the thalamus, in addition to limbic and somatic motor systems, activate central autonomic nuclei and their pathways, including those implicated in cerebrovascular regulation.
Subject(s)
Brain/metabolism , Cerebral Cortex/blood supply , Proto-Oncogene Proteins c-fos/biosynthesis , Seizures/metabolism , Animals , Autonomic Nervous System/drug effects , Autonomic Nervous System/physiology , Biomarkers , Brain/blood supply , Brain/physiopathology , Carbachol/pharmacology , Cerebral Cortex/metabolism , Cerebral Cortex/physiopathology , Electroencephalography , Entorhinal Cortex/metabolism , Entorhinal Cortex/physiology , Humans , Neurons/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Rats , Regional Blood Flow/physiology , Seizures/diagnosis , Seizures/physiopathology , Thalamus/drug effects , Thalamus/physiology , Vasodilation/drug effects , Vasodilation/physiologyABSTRACT
Convulsive seizures were elicited by a single unilateral microinjection of the cholinergic muscarinic agonist, carbachol, into the thalamus. Moreover, using systematic single microinjections of carbachol, we identified specific regions within the thalamus which were the origin of behavioural and electrocortical correlates associated with limbic and/or generalized convulsive seizures. Neither serotonin, noradrenaline nor glutamate had any convulsive effect when injected into the epileptogenic thalamic areas. The specific epileptogenic sites identified within the thalamus may provide a new experimental model which should prove useful for exploring the thalamic and thalamo-cortical mechanisms underlying limbic and generalized convulsive seizure disorders.
Subject(s)
Carbachol/pharmacology , Seizures/chemically induced , Thalamus , Animals , Brain Mapping , Limbic System , Rats , Rats, Wistar , Seizures/physiopathology , Time FactorsABSTRACT
Changes in local cerebral blood flow (lCBF) and local cerebral glucose utilization (lCGU) were assessed in dopaminergic primary target areas in the rat 6 weeks after unilateral lesion of dopaminergic neurons within the substantia nigra pars compacta (SNc) and adjacent ventrotegmental area (VTA) using 6-hydroxydopamine (6-OHDA). lCBF and lCGU were determined using the autoradiographic [14C]iodoantipyrine and [14C]2-deoxyglucose method. Dopaminergic deafferentation provoked a marked unilateral lCBF decrease in the dorso-lateral portion of the rostral caudate-putamen. The decrease in lCBF was not associated with significant changes in glucose metabolism. Thus, lesions of dopaminergic afferents to the caudate-putamen appear to provoke a sustained decrease in basal blood flow with unchanged local metabolic activity.
Subject(s)
Caudate Nucleus/metabolism , Cerebrovascular Circulation , Dopamine/analysis , Energy Metabolism , Mesencephalon/drug effects , Neurons/metabolism , Putamen/metabolism , Afferent Pathways/physiology , Animals , Caudate Nucleus/drug effects , Caudate Nucleus/pathology , Cell Death , Deoxyglucose/metabolism , Female , Mesencephalon/blood supply , Neurons/drug effects , Neurons/ultrastructure , Oxidopamine/toxicity , Putamen/drug effects , Putamen/pathology , Rats , Rats, Sprague-DawleyABSTRACT
The present study was carried out to determine whether genetically obese Zucker rats present changes in brain glucose utilization and/or insulin binding when compared to their lean counterparts. Glucose utilization in the whole brain, determined by measurement of 2-deoxy(1-3H)glucose-6-phosphate, was significantly lower in obese than in lean Zucker rats. In order to precise the structure involved, we then used quantitative autoradiography methods after either (1-14C) 2-deoxyglucose injection or 125I-insulin incubation. In obese rats, local cerebral glucose utilization (LCGU) was significantly decreased in the external plexiform layer (-37%, p < 0.05), in the lateral hypothalamus (-23%, p < 0.05), and in the basolateral amygdaloid nucleus (-30%, p < 0.05). In contrast, no difference in specific insulin binding was found between the two genotypes in any of the areas studied. These results are consistent with some data showing a decrease of LCGU in hyperinsulinemic rats. All together, these data show perturbations of glucose utilization, particularly in structures linked to the regulation of body weight and food intake in obese Zucker rats.
Subject(s)
Blood Glucose/metabolism , Body Weight/genetics , Brain/physiology , Receptor, Insulin/genetics , Amygdala/anatomy & histology , Amygdala/physiology , Animals , Arcuate Nucleus of Hypothalamus/anatomy & histology , Arcuate Nucleus of Hypothalamus/physiology , Autoradiography , Body Weight/physiology , Brain/anatomy & histology , Brain Mapping , Deoxyglucose/metabolism , Female , Hypothalamic Area, Lateral/anatomy & histology , Hypothalamic Area, Lateral/physiology , Olfactory Bulb/anatomy & histology , Olfactory Bulb/physiology , Rats , Rats, Zucker , Receptor, Insulin/physiologyABSTRACT
The effect of electrical stimulation of the centromedian-parafascicular complex on local cerebral blood flow and local cerebral glucose utilization was investigated in anesthetized, paralysed and ventilated rats. Local cerebral blood flow and local cerebral glucose utilization were measured in separate groups of animals using the autoradiographic (14C)iodoantipyrine and (14C)2-deoxyglucose methods, respectively. Because of the well-established centromedian-parafascicular complex neuroanatomical connections, three functional neuronal systems were analysed and compared: the extrapyramidal motor system the limbic system and the reticular formation, also known as the ascending activating system. Cortical regions not included in the limbic system were considered separately. The validity of comparisons between changes in local cerebral blood flow and local cerebral glucose utilization across the brain was verified by assessing the reactivity and stability of the cortical blood flow during long-term centromedian-parafascicular complex stimulation. Centromedian-parafascicular complex stimulation elicited a marked but heterogeneous increase in local cerebral blood flow in 50 of the 52 cerebral structures measured. The most pronounced increases were seen in the lateral habenular nucleus (331 +/- 30% of control), the zona incerta (400 +/- 55%), the mesencephalic reticular formation (415 +/- 122%) and the parietal cortex (211 +/- 35%). In contrast, local cerebral glucose utilization remained statistically unchanged (P greater than 0.05) in 28 of these 50 individual brain regions during centromedian-parafascicular complex stimulation. The most pronounced increases in local cerebral glucose utilization were seen in the zona incerta (123 +/- 28%) and the mesencephalic reticular formation (193 +/- 26%). Local cerebral blood flow and local cerebral glucose utilization were linearly related in unstimulated controls, considering either all brain regions taken as a whole or the three systems separately. The significant increase in the slopes of the regression line between local cerebral blood flow and local cerebral glucose utilization for the reticular formation and the limbic system during centromedian-parafascicular complex stimulation indicates, however, that the coupling mechanisms for these systems, but not for the extrapyramidal motor system, were reset. The local cerebral blood flow to local cerebral glucose utilization ratio was heterogeneous in controls and differentially increased during centromedian-parafascicular complex stimulation, being markedly pronounced in the parietal cortex and in the reticular formation. We conclude that these results, for the first time, provide evidence that, the functionally well-defined neural networks may have different mechanisms whereby changes in vascular and metabolic demands are regulated.
Subject(s)
Cerebrovascular Circulation/physiology , Thalamic Nuclei/physiology , Anesthesia , Animals , Autoradiography , Blood Gas Analysis , Brain Chemistry/drug effects , Electric Stimulation , Glucose/metabolism , Mass Spectrometry , Neurons/physiology , Rats , Rats, WistarABSTRACT
Changes in cerebral cortical perfusion (CBFLDF), local cerebral blood flow (lCBF) and local cerebral glucose utilization (lCGU) elicited by unilateral cortical spreading depression (SD) were monitored and measured in separate groups of rats anesthetized with alpha-chloralose. CBFLDF was recorded with laser Doppler flowmetry, while lCBF and lCGU were measured by the quantitative autoradiographic [14C]iodoantipyrine and [14C]-2-deoxyglucose methods, respectively. SD elicited a wave of hyperemia after a latency of 2 to 3 min followed by an oligemic phase. Ninety minutes following the onset of SD cortical (frontal, parietal and occipital) lCBF and lCGU were essentially the same as on the contralateral side and in sham-treated rats. However, alteration in the lCBF and lCGU in upper and lower brainstem persisted. The present results demonstrate, for the first time, that long-lasting cerebrovascular and metabolic alterations take place within the subcortical regions following SD. These regions provide an attractive site to integrate observations in man concerning spreading depression and the aura of migraine with the other features of the syndrome.
Subject(s)
Cerebrovascular Circulation/physiology , Cortical Spreading Depression/physiology , Animals , Autoradiography , Brain Injuries/diagnosis , Brain Injuries/metabolism , Brain Injuries/physiopathology , Migraine Disorders/physiopathology , Rats , Rats, Inbred StrainsABSTRACT
Cessation of chronic (5 days), unilateral infusion of GABA into the somatomotor cortex of rats induces focal epileptic spikes which remain limited to the infused site and never evolve into generalized seizures. We have considered this finding as a new model of focal epilepsy and named it "GABA withdrawal syndrome". In the present study, we have measured local cerebral glucose utilization in order to map the cortical and subcortical regions involved in the GABA withdrawal syndrome. Local cerebral glucose utilization increased two- to three-fold in a 1-1.5 mm diameter area, involving all the cortical layers at the GABA-infusion site. This hypermetabolic area contained a central (1-2 mm diameter) hypometabolic zone showing neuronal depopulation in some animals. Except for the epileptic focus, the hemisphere ipsilateral to the infusion site was slightly hypometabolic. However, there was a large increase (three- to five-fold) in some ipsilateral thalamic nuclei (posterior oralis, ventralis postero-lateralis, centralis lateralis, ventralis lateralis and reticularis thalami nucleus). The local cerebral glucose utilization of the contralateral cortex and thalamus were unchanged. The present results confirm the focal nature of the epileptogenic syndrome produced by stopping chronic, intracortical GABA infusion. These results are markedly different from those described in the penicillin focal epilepsy model. Our data also show that specific ipsilateral thalamic relays may, by an as yet unknown mechanism, play a role in maintaining paroxysmal activity during the GABA withdrawal syndrome.
Subject(s)
Brain/physiopathology , Cerebral Cortex/physiopathology , Epilepsies, Partial/physiopathology , gamma-Aminobutyric Acid/pharmacology , Animals , Brain/anatomy & histology , Cerebral Cortex/drug effects , Cerebral Cortex/physiology , Deoxyglucose/metabolism , Drug Administration Schedule , Electroencephalography , Infusions, Parenteral , Kinetics , Male , Organ Specificity , Rats , Rats, Inbred Strains , Reference Values , gamma-Aminobutyric Acid/administration & dosageABSTRACT
The present study was carried out to characterize the effects of insulin, using the euglycemic hyperinsulinemic clamp, on insulin binding and glucose utilization in specific areas of rat brain, by autoradiographic methods. Binding of [125I]Insulin was significantly higher in the hippocampus CA1, the ventromedial and lateral hypothalamus nuclei of the hyperinsulinemic rats than in control rats. Glucose utilization was slightly but not significantly decreased in the hippocampus CA1, the ventromedial and lateral hypothalamus of hyperinsulinemic rats. These data suggest that insulin, via its specific receptors, may exert its central actions by affecting glucose utilization.
Subject(s)
Brain/metabolism , Glucose/pharmacokinetics , Hyperinsulinism/metabolism , Insulin/metabolism , Animals , Brain/drug effects , Brain/physiopathology , Insulin/pharmacokinetics , Male , Rats , Rats, Inbred StrainsABSTRACT
Electrical stimulation of the centromedian-parafascicular complex (CM-Pf) in anesthetized (chloralose) and paralyzed (tubocurarine) rats elicits a widespread cerebrovascular dilatation. Regional cerebral blood flow (rCBF) was measured in dissected tissue samples of 10 brain regions (medulla, pons, cerebellum, inferior colliculus, superior colliculus, frontal parietal and occipital cortices, caudate-putamen and corpus callosum) by [14C]iodoantipyrine method. In unstimulated and sham-operated rats rCBF ranged from 40 +/- 3 (ml/100 g/min) in corpus callosum to 86 +/- 6 (ml/100 g/min) in inferior colliculus. During CM-Pf stimulation, rCBF increased significantly (P less than 0.05, analysis of variance and Scheffe's test) in all cerebral regions bilaterally ranging from +118% in parietal cortex to +38% in cerebellum. Although cerebral vasodilation elicited by CM-Pf stimulation persisted after unilateral transection of the cervical sympathetic trunk, the cortical CBF was significantly reduced (P less than 0.05) on the denervated side. Acute adrenalectomy significantly (P less than 0.05) decreased elevated rCBF during CM-Pf stimulation in all cortical regions (frontal-36%, parietal -34%, and occipital -27%) and in caudate nucleus (-37%). Thus, excitation of neurons originating in, or fibers passing through the CM-Pf can elicit a powerful cerebral vasodilation. The cerebral vasodilation is modulated by cervical sympathectomy and circulating adrenal hormones. We conclude that CM-Pf elicited vasodilation is at least partly mediated by intrinsic neural pathways.
Subject(s)
Cerebrovascular Circulation , Thalamic Nuclei/physiology , Adrenalectomy , Animals , Blood Pressure , Electric Stimulation , Functional Laterality/physiology , Neural Pathways/physiology , Rats , Rats, Inbred Strains , SympathectomySubject(s)
Calcification, Physiologic/drug effects , Gastrointestinal Motility/drug effects , Hydroxycholecalciferols/pharmacology , Vitamin D Deficiency/metabolism , Animals , Bone and Bones/metabolism , Calcium/metabolism , Dihydroxycholecalciferols/pharmacology , Isomerism , Male , Phosphates/blood , Rats , Structure-Activity RelationshipABSTRACT
The separation of 5-cholestene-3 beta, 25 (RS), 25-triol 3,26-diacetate into the diastereoisomers 25R and 25S by means of high pressure liquid chromatography (HPLC) is described. Their absolute configuration cannot be yet established. The less polar diastereoisomer is arbitrarily called 25 zeta1 and the more polar one 25 zeta2. Bromination, dehydrobromination and ultraviolet irradiation conducted to 25 zeta1, 26(OH2D3 and 25 zeta2, 26(OH)2D3 respectively. Their biological activity is described.
Subject(s)
Dihydroxycholecalciferols , Dihydroxycholecalciferols/isolation & purification , Hydroxycholecalciferols , Animals , Bone and Bones/drug effects , Bone and Bones/metabolism , Calcium/metabolism , Dihydroxycholecalciferols/pharmacology , Hydroxycholecalciferols/isolation & purification , Magnetic Resonance Spectroscopy , Male , Rats , Stereoisomerism , Structure-Activity RelationshipABSTRACT
The resolution of 5-cholestene-3 beta, 24 RS,25 triol 3,24-diacetate into the diastereoisomers 24 R and 24 S by means of liquid chromatography is described. Bromination, dehydrobromination and ultraviolet irradiation of both diastereoisomers led to 24 R,25 (OH)2D3 and 24 S,25 (OH)2D3 respectively. Their structure was further confirmed by thin layer chromatography, ultraviolet and mass spectroscopy.