Localization and Characterization of Binding Sites with High Affinity for [<sup>3</sup>H]Ouabain in Cerebral Cortex of Rabbit Brain Using Quantitative Autoradiography

Antonelli, Marta C.; Baskin, Denis G.; Garland, Miriam; Stahl, William L.

doi:10.1111/j.1471-4159.1989.tb10916.x

Cited by 23 publications

(16 citation statements)

References 33 publications

(22 reference statements)

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“…The analysis of cerebral cortex data (rat and rabbit brain) was done in laminae 3-4. These laminae were previously reported to show the highest ouabain binding levels in rabbit cerebral cortex (Antonelli et al, 1989). This is in agreement with previous findings showing that the highest Na+ ,K+-ATPase activity is found in lamina 4 (Stahl and Broderson, 1976;Broderson et al, 1978).…”

Section: Resultssupporting

confidence: 93%

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Antonelli

Casillas

Arnaiz

1991

J of Neuroscience Research

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There is growing evidence on the existence of endogenous ouabain-like factors that modulate Na+, K(+)-ATPase activity. In this laboratory, two soluble subfractions (peaks I and II) were previously separated from rat cerebral cortex, which had opposite effects on Na+, K(+)-ATPase activity. Peak I stimulated and peak II inhibited the enzyme (Rodríguez de Lores Arnaiz and Antonelli de Gómez de Lima, Neurochem Res 11:933-947, 1986). The same effects are now reported for K(+)-p-nitrophenyphosphatase activity. Localization of high-affinity ouabain binding in rat brain was done by quantitative autoradiography using a microcomputer digital imaging system. Peak I did not modify, whereas peak II blocked ouabain binding in areas 3-4 of cerebral cortex, dentate gyrus, stria terminalis, thalamic nuclei, and basal ganglia. Similar results were obtained when ouabain binding was determined in rabbit cerebral cortex and by a conventional filtration assay in nerve ending membranes obtained from rat cerebral cortex. These results favour the idea that the factor present in peak II fraction might behave as an ouabain-like substance.

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Section: Resultssupporting

confidence: 93%

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Antonelli

Casillas

Arnaiz

1991

J of Neuroscience Research

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“…It is clear that both FS interneurons and PYR1 neurons have more active resting Na + –K + ATPase activity than PYR2 neurons. However, only a portion of the total Na + –K + ATPase molecules are phosphorylated and thus active at rest and sensitive to pharmacological blockade (Forbush & Hoffman, 1979; Antonelli et al 1989). To test the Na + –K + ATPase capacity of the different cell groups we induced Na + –K + ATPase activity by intracellularly loading cells with Na + using two methods.…”

Section: Resultsmentioning

confidence: 99%

Differential effects of Na⁺–K⁺ ATPase blockade on cortical layer V neurons

Anderson

Huguenard

Prince

2010

The Journal of Physiology

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Sodium-potassium ATPase ('Na+ -K + ATPase') contributes to the maintenance of the resting membrane potential and the transmembrane gradients for Na + and K + in neurons. Activation of Na + -K + ATPase may be important in controlling increases in intracellular sodium during periods of increased neuronal activity. Down-regulation of Na + -K + ATPase activity is implicated in numerous CNS disorders, including epilepsy. Although Na + -K + ATPase is present in all neurons, little is known about its activity in different subclasses of neocortical cells. We assessed the physiological properties of Na in fast-spiking (FS) interneurons and pyramidal (PYR) cells to test the hypothesis that Na+ -K + ATPase activity would be relatively greater in neurons that generated high frequency action potentials (the FS cells). Whole-cell patch clamp recordings were made from FS and PYR neurons in layer V of rat sensorimotor cortical slices maintained in vitro using standard techniques. Bath perfusion of Na + -K + ATPase antagonists (ouabain or dihydro-ouabain) induced either a membrane depolarization in current clamp, or inward current under voltage clamp in both cell types. PYR neurons were divided into two subpopulations based on the amplitude of the voltage or current shift in response to Na + -K + ATPase blockade. The two PYR cell groups did not differ significantly in electrophysiological properties including resting membrane potential, firing pattern, input resistance and capacitance. Membrane voltage responses of FS cells to Na + -K + ATPase blockade were intermediate between the two PYR cell groups (P < 0.05). The resting Na + -K + ATPase current density in FS interneurons, assessed by application of blockers, was 3-to 7-fold larger than in either group of PYR neurons. Na + -K + ATPase activity was increased either through direct Na + loading via the patch pipette or by focal application of glutamate (20 mm puffs). Under these conditions FS interneurons exhibited the largest increase in Na + -K + ATPase activity. We conclude that resting Na + -K + ATPase activity and sensitivity to changes in internal Na + concentration vary between and within classes of cortical neurons. These differences may have important consequences in pathophysiological disorders associated with down-regulation of Na + -K + ATPase and hyperexcitability within cortical networks.

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“…Brains were rapidly frozen by immersion in Freon (À408C) and stored at À808C. Frozen coronal sections (12 lm) were cut at five different anatomical levels in a cryostat at À208C, thawed, mounted onto gelatin-coated microscopic slides, and stored at À808C until use (Antonelli et al, 1989).…”

Section: Preparation Of Brain Sectionsmentioning

confidence: 99%

Morphine withdrawal syndrome and its prevention with baclofen: Autoradiographic study of μ‐opioid receptors in prepubertal male and female mice

Diaz

Barros

Antonelli

et al. 2006

Synapse

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Although the expression of the morphine (MOR) withdrawal syndrome is more marked in male mice than in females, we have demonstrated that the GABAB agonist baclofen (BAC) is able to attenuate MOR withdrawal signs in either sex. In order to extend these previous observations, the aim of the present study was to evaluate the mu-opioid receptor labeling in various brain areas in mice of either sex, during MOR withdrawal and its prevention with BAC. Prepubertal Swiss-Webster mice were rendered dependent by intraperitonial (i.p.) injection of MOR (2 mg/kg) twice daily for 9 days. On the 10th day, dependent animals received naloxone (NAL; 6 mg/kg, i.p.) 60 min after MOR, and another pool of dependent mice received BAC (2 mg/kg, i.p.) previous to NAL. Thirty minutes after NAL, mice were sacrificed and autoradiography with [3H]-[D-Ala2, N-Me-Phe4, -glycol5] enkephalin (DAMGO) was carried out on mice brains at five different anatomical levels. Autoradiographic mapping showed a significant increase of mu-opioid receptor labeling during MOR withdrawal in nucleus accumbens core (NAcC), caudate putamen (CPu), mediodorsal thalamic nucleus (MDTh), basolateral and basomedial amygdala, and ventral tegmental area vs. respective control groups in male mice. In contrast, opiate receptor labeling was not significantly modified in any of the brain areas studied in withdrawn females. BAC reestablished mu-opioid receptor binding sites during MOR withdrawal only in NAcC of males, and a similar tendency was observed in CPu and MDTh, even when it was not statistically significant. The sexual dimorphism observed in the present study confirms previous reports indicating a greater sensitivity of males in response to MOR pharmacological properties. The present results suggest that the effect of BAC in preventing the expression of MOR withdrawal signs could be related with the ability of BAC to reestablish the mu-opioid receptor labeling in certain brain areas.

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Localization and Characterization of Binding Sites with High Affinity for [³H]Ouabain in Cerebral Cortex of Rabbit Brain Using Quantitative Autoradiography

Cited by 23 publications

References 33 publications

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Differential effects of Na⁺–K⁺ ATPase blockade on cortical layer V neurons

Morphine withdrawal syndrome and its prevention with baclofen: Autoradiographic study of μ‐opioid receptors in prepubertal male and female mice

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Localization and Characterization of Binding Sites with High Affinity for [3H]Ouabain in Cerebral Cortex of Rabbit Brain Using Quantitative Autoradiography

Cited by 23 publications

References 33 publications

Effect of Na+, K+‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Effect of Na+, K+‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Differential effects of Na+–K+ ATPase blockade on cortical layer V neurons

Morphine withdrawal syndrome and its prevention with baclofen: Autoradiographic study of μ‐opioid receptors in prepubertal male and female mice

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Localization and Characterization of Binding Sites with High Affinity for [³H]Ouabain in Cerebral Cortex of Rabbit Brain Using Quantitative Autoradiography

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Effect of Na⁺, K⁺‐ATPase modifiers on high‐affinity ouabain binding determined by quantitative autoradiography

Differential effects of Na⁺–K⁺ ATPase blockade on cortical layer V neurons