Glial origin of negative shifts of cortical surface potential during tetanic stimulation: Microelectrode study and mathematical analysis

Experiments on anesthetized cats showed that a negative shift of potential on the surface of the cerebral cortex caused by its tetanic stimulation is similar in shape and time course to the depolarization shift of membrane potential of the glial cells, but has a more rapid decline. The hyperpolarization shifts of membrane potential of neurons differed in shape and time course from the negative shift of cortical surface potential. It is concluded that the contribution of hyperpolarization of neurons to the surface-negative potential shift during tetanic stimulation may be manifested visibly only at the beginning (the first 200–300 msec) of such stimulation. The negative potential shift on the cortical surface is due mainly to depolarization of glial cells under the influence of K⁺ secreted from excited nerve cells.     

Facilitation of reactions of cortical neurons to cortical and peripheral stimulation after tetanization of the brain surface

The electrical activity of neurons of the sensomotor cortex of an unanesthetized rabbit was investigated. Conditioning tetanization of the cortex was carried out through surface electrodes located close to the site of the lead. Test stimuli were supplied through electrodes 2.5–12 mm more caudally on the cortical surface. In addition, peripheral test stimuli were applied. Impulse reactions to previously ineffective stimuli develop after conditioning tetanization and prolonged (up to 1 min) intensification of exciting postsynaptic potentials (EPSP) to cortical and peripheral test stimuli is observed. Facilitation of the reactions is especially clear during tetanization superthreshold for evoking epileptiform afterdischarges. It continued after the conclusion of these discharges and could also be observed during tetanization subthreshold for evoking afterdischarges. The time course of the facilitation was similar to the time course of the post-tetanic intensification of reactions of single stimuli applied with the electrodes used for tetanization. An analysis of the changes in intracellular activity makes it possible to assume that the mechanism of post-tetanic potentiation (PTP) lies at the basis of the described facilitation, which is considered as the "cellular analog" of the dominant focus which develops as a result of tetanization of the cortical surface.Institute of the Brain, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 3, No. 1, pp. 3–12, January–February, 1971.     

Integrative properties of the microsystem of cortical neurons during repeated local stimulation

Small groups of neighbouring neurons in neocortex are able to form separate microsystem in model situation of habituation during the local repetitive action of acetylcholine. This new functional unit exhibits a number of main properties which are characteristic of systemic processes of habituation. Only some of the properties don't reproduce in microsystem of cortical neurons. There is no effect of full extinction of responses and no direct relationship between the decrease of speed quantity of responses and stimulus action frequency. The data obtained are considered as significant argument for identification of special intermediate level of integrative processes--microsystem level.     

Nitrogen shuttling between neurons and glial cells during glutamate synthesis

The relationship between neuronal glutamate turnover, the glutamate/glutamine cycle and de novo glutamate synthesis was examined using two different model systems, freshly dissected rat retinas ex vivo and in vivo perfused rat brains. In the ex vivo rat retina, dual kinetic control of de novo glutamate synthesis by pyruvate carboxylation and transamination of alpha-ketoglutarate to glutamate was demonstrated. Rate limitation at the transaminase step is likely imposed by the limited supply of amino acids which provide the alpha-amino group to glutamate. Measurements of synthesis of (14)C-glutamate and of (14)C-glutamine from H(14)CO(3) have shown that (14)C-amino acid synthesis increased 70% by raising medium pyruvate from 0.2 to 5 mM. The specific radioactivity of (14)C-glutamine indicated that approximately 30% of glutamine was derived from (14)CO(2) fixation. Using gabapentin, an inhibitor of the cytosolic branched-chain aminotransferase, synthesis of (14)C-glutamate and (14)C-glutamine from H(14)CO(3)(-) was inhibited by 31%. These results suggest that transamination of alpha-ketoglutarate to glutamate in Müller cells is slow, the supply of branched-chain amino acids may limit flux, and that branched-chain amino acids are an obligatory source of the nitrogen required for optimal rates of de novo glutamate synthesis. Kinetic analysis suggests that the glutamate/glutamine cycle accounts for 15% of total neuronal glutamate turnover in the ex vivo retina. To examine the contribution of the glutamate/glutamine cycle to glutamate turnover in the whole brain in vivo, rats were infused intravenously with H(14)CO(3)(-). (14)C-metabolites in brain extracts were measured to determine net incorporation of (14)CO(2) and specific radioactivity of glutamate and glutamine. The results indicate that 23% of glutamine in the brain in vivo is derived from (14)CO(2) fixation. Using published values for whole brain neuronal glutamate turnover, we calculated that the glutamate/glutamine cycle accounts for approximately 60% of total neuronal turnover. Finally, differences between glutamine/glutamate cycle rates in these two model systems suggest that the cycle is closely linked to neuronal activity.     

Steady potential and extracellular potassium shifts in the rat neocortex during sustained low-frequency electrical stimulation of the cortical surface

Cyclic excitation arising in the rat neocortex under the influence of low-frequency electrical stimulation was studied. The duration of the excitation cycles and intervals between them depended on the stimulation parameters. In the period of excitation the negative shift of steady potential reached 4–5 mV, and the extracellular potassium ion concentration, measured with the aid of potassium-selective microelectrodes, rose to 8–10 mM. The process of periodic excitation was not self-maintained and it ceased after the current was stopped. As a result of small doses of pentobarbital (10–20 mg/kg) the response thresholds rose, and intervals between excitation phases were increased by several times. Pentobarbital in a dose of 30–40 mg/kg completely abolished cyclic excitation for 2–3 h. Correlation between shifts of steady potential and extracellular potassium concentration differed in anesthetized and unanesthetized animals.     

Asymmetric inheritance of radial glial fibers by cortical neurons

Recent studies demonstrated the neuronogenic role of radial glial cells (RGCs) in the rodent. To reveal the fate of radial glial processes, we intensively monitored divisions of RGCs in DiI-labeled slices from the embryonic day 14 mouse cortex. During RGC division, each pia-connected fiber becomes thin but is neither lost nor divided; it is inherited asymmetrically by one daughter cell. In divisions that produce a neuron and a progenitor, the neuron inherits the pial fiber, also grows a thick ventricular process for several hours, and is therefore indistinguishable from the progenitor RGC. The ventricular process in the radial glial-like neuron ("radial neuron") then collapses, leading to ascent of the neuron by using the "recycled" radial fiber.     

Plasticity and its physiological markers in the microsystem of cortical neurons during repeated local stimulation

Separate neuronal microsystems in the sensomotor cerebral cortex are able to exhibit the functional plasticity under conditions of repeated action of acetylcholine applied microiontophoretically. The characteristic properties of dynamics in activity of single cortex neurones are determined mainly by the initial reactivity to transmitter and by the state of nervous cells of the surrounding microsystem. The composition and succession of response components as well the duration of excitatory stage of reaction to acetylcholine serve as physiological markers of plastic properties of neurones in cortical microsystem.     

Responses of cortical neurons to viscero-somatic stimulation

Responses of cortical neurons in the posterior sigmoid gyrus of cats anesthetized with chloralose to electrical stimulation of somatic and visceral nerves were recorded. Bimodal viscero-somatic neurons are predominant in this part of the cortex and some of them also respond to light. Besides the polysensory modally-specific neurons it was also possible to distinguish a group of modally nonspecific cells (27%), whose responses to different stimuli did not differ statistically from each other. Simultaneous stimulation of visceral and somatic nerves led to facilitation of activity of the long-latency neurons; this was reflected in a decrease of 10 msec in the latent period of the response and an increase in the number of spikes per discharge.A. A. Zhdanov Leningrad University. Tadjik University, Dushanbe. Translated from Neirofiziologiya, Vol. 3, No. 6, pp. 574–581, November–December, 1971.     

Conditions of appearance of off responses of sensomotor cortical neurons to photic stimulation in cats

In acute experiments on cats under chloralose anesthesia (70 mg/kg) unit activity was recorded extra- and intracellularly in the sensomotor cortex (areas 4 and 6) during prolonged (up to 1000 msec) photic stimulation. Responses of on-off type were generated by 100% of neurons tested to photic stimuli whose duration corresponded to the recovery cycle of functional changes after a single flash, determined by the paired stimulation method. Cutaneous stimulation affected the appearance of the photic off response if it led to a spike discharge of the neuron before the off response. It is suggested that IPSPs of cortical neurons largely determine both the duration of the cycle of functional recovery after a single flash and also differences in the pattern of generation of the off response and its interaction with responses to cutaneous stimulation.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 9, No. 4, pp. 355–360, July–August, 1977.     

Cholesterol homeostasis in neurons and glial cells

Cholesterol is highly enriched in the brain compared to other tissues. Essentially all cholesterol in the brain is synthesized endogenously since plasma lipoproteins are unable to cross the blood-brain barrier. Cholesterol is transported within the central nervous system in the form of apolipoprotein E-containing lipoprotein particles that are secreted mainly by glial cells. Cholesterol is excreted from the brain in the form of 24-hydroxycholesterol. Apolipoprotein E and cholesterol have been implicated in the formation of amyloid plaques in Alzheimer's disease. In addition, the progressive neurodegenerative disorder Niemann-Pick C disease is characterized by defects in intracellular trafficking of cholesterol.     

Leaching of concanavalin A during affinity chromatographic isolation of cell surface glycoproteins from human fetal neurons and glial cells.

Preparation of surface glycoproteins from human fetal brain cells by affinity chromatography on Con A-Sepharose 4B was a problematic endeavor due to leaching of Con A from the matrix. Dissociation of Con A from the matrix took place irrespective of the presence of lipid and/or detergent and the buffer composition during chromatography and was apparently related to the nature of the protein under study. Pretreatment of Con A-Sepharose with 6 M guanidine or 8 M urea reduced Con A leaching. The Con A eluate also contained noncovalently associated glycolipid. Elution at 25 degrees C rendered fractions containing a higher degree of Con A and glycolipid contamination compared to the negligible contamination by these two components when elution was carried out at 4 degrees C. This phenomenon was attributed to the formation of heterogeneous mixed micelles of glycoprotein.     

Spontaneous activity and evoked responses of cerebellar cortical neurons to vagal and limb nerve stimulation in pigeons

Spontaneous activity and evoked reponses of the cerebellar cortical neurons of decerebrate pigeons to stimulation of the vagus nerve and nerves of the limbs were investigated. The cell responses were uniform in type regardless of the character of stimulation. Phasic and tonic responses, predominantly of excitatory type, were recorded. The phasic responses had both short (10–20 msec) and long (up to 80 msec) latent periods. The latter predominated. Features distinguishing the unit responses to vagal stimulation of the limb nerves included: a smaller number of activated cells, longer latent period of the responses, absence of activation of Purkinje cells through the climbing fibers, and lower capacity for rhythm binding.     

Modifications in energy metabolism during the development of chick glial cells and neurons in culture

Developmental changes in lactate dehydrogenase (LDH), enolase, hexokinase (HK), malate dehydrogenase (MDH), and glutamate dehydrogenase (GDH) activities were measured in cultures of pure neurons and glial cells prepared from brains of chick embryos (8 day-old for neurons, 14 day-old for glial cells) as a function of cellular development with time in culture. The modifications observed in culture were compared to those measured in brain extracts during the development of the nervous tissue in the chick embryo and during the post-hatching period. A significant increase of MDH, GDH, LDH, and enolase activities are observed in neurons between 3 and 6 days of culture, whereas simultaneously a decrease of HK values occurs. In the embryonic brain between 11 and 14 days of incubation, which would correspond for the neuronal cultures to day 3 through 6, modifications of MDH, GDH, HK, and enolase levels are similar to those observed in neurons in culture. Only the increase of LDH activity is less pronounced in vivo than in cultivated cells. The evolution of the tested enzymatic activities in the brain of the chick during the period between 7 days before and 10 days after hatching is quite similar to that observed in cultivated glial cells (prepared from 14 day-old embryos) between 6 and 18 days of culture. All tested activities increased in comparable proportions. The modifications of the enzymatic profile indicate that some maturation phenomena affecting energy metabolism of neuronal and glial elements in culture, are quite similar to those occuring in the total nervous tissue. A relationship between the development of the energy metabolism of the brain and differentiation processes affecting neuroblasts and the glial-forming cells is discussed.     

Impulse activity of the bulbar cardiovascular neurons during myocardial ischemia and stimulation of the cortical sensorimotor zone

In acute experiments on cats performed under nembutal anesthesia the stimulation of sensorimotor zone in cerebral hemisphere cortex changed the impulse activity of interneurons of bulbar cardiovascular centre and not of the afferent neurons. The analysis of the activity of afferent neurons and interneurons has shown a decrease in coordination between the reaction of these cells to the development of ischemic myocardial lesions during the cortex stimulation. In these conditions bulbar cardiovascular neurons could both increase and decrease the impulse activity. These changes seem to be the reason for the growing incidence of idioventricular ischemic arrhythmias during cortical stimulation.