Electrophysiological recordings and calcium measurements in striatal large aspiny interneurons in response to combined O2/glucose deprivation

Antonio Pisani, Paolo Calabresi, Diego Centonze, Girolama A. Marfia, Giorgio Bernardi

Research output: Contribution to journalArticle

Abstract

The effects of combined O2/glucose deprivation were investigated on large aspiny (LA) interneurons recorded from a striatal slice preparation by means of simultaneous electrophysiological and optical recordings. LA interneurons were visually identified and impaled with sharp microelectrodes loaded with the calcium (Ca2+)-sensitive dye bis-fura-2. These cells showed the morphological, electrophysiological, and pharmacological features of large striatal cholinergic interneurons. O2/glucose deprivation induced a membrane hyperpolarization coupled to a concomitant increase in intracellular Ca2+ concentration ([Ca2+](i)). Interestingly, this [Ca2+](i) elevation was more pronounced in dendritic branches rather than in the somatic region. The O2/glucose-deprivation-induced membrane hyperpolarization reversed its polarity at the potassium (K+) equilibrium potential. Both membrane hyperpolarization and [Ca2+](i) rise were unaffected by TTX or by a combination of ionotropic glutamate receptors antagonists, D-2 amino-5- phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione. Sulfonylurea glibenclamide, a blocker of ATP-sensitive K+ channels, markedly reduced the O2/glucose-deprivation-induced membrane hyperpolarization but failed to prevent the rise in [Ca2+](i). Likewise, charybdotoxin, a large K+ channel (BK) inhibitor, abolished the membrane hyperpolarization but did not produce detectable changes of [Ca2+](i) elevation. A combination of high-voltage- activated Ca2+ channel blockers; significantly reduced both the membrane hyperpolarization and the rise in [Ca2+](i). In a set of experiments performed without dye in the recording electrode, either intracellular bis- (o-aminophenoxy)-N,N,N',N'-tetraacetic acid or external barium abolished the membrane hyperpolarization induced by O2/glucose deprivation. The hyperpolarizing effect on membrane potential was mimicked by oxotremorine, an M2-like muscarinic receptor agonist, and by baclofen, a GABA(B) receptor agonist. However, this membrane hyperpolarization was not coupled to an increase but rather to a decrease of the basal [Ca2+](i). Furthermore glibenclamide did not reduce the oxotremorine- and baclofen-induced membrane hyperpolarization. In conclusion, the present results suggest that in striatal LA cells, O2/glucose deprivation activates a membrane hyperpolarization that does not involve ligand-gated K+ conductances but is sensitive to barium, glibenclamide, and charybdotoxin. The increase in [Ca2+](i) is partially due to influx through voltage-gated high-voltage- activated Ca2+ channels.

Original languageEnglish
Pages (from-to)2508-2516
Number of pages9
JournalJournal of Neurophysiology
Volume81
Issue number5
Publication statusPublished - 1999

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Corpus Striatum
Interneurons
Calcium
Glucose
Membranes
Glyburide
Charybdotoxin
Oxotremorine
Barium
Coloring Agents
GABA-B Receptor Agonists
GABA-B Receptors
Large-Conductance Calcium-Activated Potassium Channels
6-Cyano-7-nitroquinoxaline-2,3-dione
2-Amino-5-phosphonovalerate
Ionotropic Glutamate Receptors
Muscarinic Agonists
Excitatory Amino Acid Antagonists
Baclofen
Fura-2

ASJC Scopus subject areas

  • Physiology
  • Neuroscience(all)

Cite this

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title = "Electrophysiological recordings and calcium measurements in striatal large aspiny interneurons in response to combined O2/glucose deprivation",
abstract = "The effects of combined O2/glucose deprivation were investigated on large aspiny (LA) interneurons recorded from a striatal slice preparation by means of simultaneous electrophysiological and optical recordings. LA interneurons were visually identified and impaled with sharp microelectrodes loaded with the calcium (Ca2+)-sensitive dye bis-fura-2. These cells showed the morphological, electrophysiological, and pharmacological features of large striatal cholinergic interneurons. O2/glucose deprivation induced a membrane hyperpolarization coupled to a concomitant increase in intracellular Ca2+ concentration ([Ca2+](i)). Interestingly, this [Ca2+](i) elevation was more pronounced in dendritic branches rather than in the somatic region. The O2/glucose-deprivation-induced membrane hyperpolarization reversed its polarity at the potassium (K+) equilibrium potential. Both membrane hyperpolarization and [Ca2+](i) rise were unaffected by TTX or by a combination of ionotropic glutamate receptors antagonists, D-2 amino-5- phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione. Sulfonylurea glibenclamide, a blocker of ATP-sensitive K+ channels, markedly reduced the O2/glucose-deprivation-induced membrane hyperpolarization but failed to prevent the rise in [Ca2+](i). Likewise, charybdotoxin, a large K+ channel (BK) inhibitor, abolished the membrane hyperpolarization but did not produce detectable changes of [Ca2+](i) elevation. A combination of high-voltage- activated Ca2+ channel blockers; significantly reduced both the membrane hyperpolarization and the rise in [Ca2+](i). In a set of experiments performed without dye in the recording electrode, either intracellular bis- (o-aminophenoxy)-N,N,N',N'-tetraacetic acid or external barium abolished the membrane hyperpolarization induced by O2/glucose deprivation. The hyperpolarizing effect on membrane potential was mimicked by oxotremorine, an M2-like muscarinic receptor agonist, and by baclofen, a GABA(B) receptor agonist. However, this membrane hyperpolarization was not coupled to an increase but rather to a decrease of the basal [Ca2+](i). Furthermore glibenclamide did not reduce the oxotremorine- and baclofen-induced membrane hyperpolarization. In conclusion, the present results suggest that in striatal LA cells, O2/glucose deprivation activates a membrane hyperpolarization that does not involve ligand-gated K+ conductances but is sensitive to barium, glibenclamide, and charybdotoxin. The increase in [Ca2+](i) is partially due to influx through voltage-gated high-voltage- activated Ca2+ channels.",
author = "Antonio Pisani and Paolo Calabresi and Diego Centonze and Marfia, {Girolama A.} and Giorgio Bernardi",
year = "1999",
language = "English",
volume = "81",
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journal = "Journal of Neurophysiology",
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T1 - Electrophysiological recordings and calcium measurements in striatal large aspiny interneurons in response to combined O2/glucose deprivation

AU - Pisani, Antonio

AU - Calabresi, Paolo

AU - Centonze, Diego

AU - Marfia, Girolama A.

AU - Bernardi, Giorgio

PY - 1999

Y1 - 1999

N2 - The effects of combined O2/glucose deprivation were investigated on large aspiny (LA) interneurons recorded from a striatal slice preparation by means of simultaneous electrophysiological and optical recordings. LA interneurons were visually identified and impaled with sharp microelectrodes loaded with the calcium (Ca2+)-sensitive dye bis-fura-2. These cells showed the morphological, electrophysiological, and pharmacological features of large striatal cholinergic interneurons. O2/glucose deprivation induced a membrane hyperpolarization coupled to a concomitant increase in intracellular Ca2+ concentration ([Ca2+](i)). Interestingly, this [Ca2+](i) elevation was more pronounced in dendritic branches rather than in the somatic region. The O2/glucose-deprivation-induced membrane hyperpolarization reversed its polarity at the potassium (K+) equilibrium potential. Both membrane hyperpolarization and [Ca2+](i) rise were unaffected by TTX or by a combination of ionotropic glutamate receptors antagonists, D-2 amino-5- phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione. Sulfonylurea glibenclamide, a blocker of ATP-sensitive K+ channels, markedly reduced the O2/glucose-deprivation-induced membrane hyperpolarization but failed to prevent the rise in [Ca2+](i). Likewise, charybdotoxin, a large K+ channel (BK) inhibitor, abolished the membrane hyperpolarization but did not produce detectable changes of [Ca2+](i) elevation. A combination of high-voltage- activated Ca2+ channel blockers; significantly reduced both the membrane hyperpolarization and the rise in [Ca2+](i). In a set of experiments performed without dye in the recording electrode, either intracellular bis- (o-aminophenoxy)-N,N,N',N'-tetraacetic acid or external barium abolished the membrane hyperpolarization induced by O2/glucose deprivation. The hyperpolarizing effect on membrane potential was mimicked by oxotremorine, an M2-like muscarinic receptor agonist, and by baclofen, a GABA(B) receptor agonist. However, this membrane hyperpolarization was not coupled to an increase but rather to a decrease of the basal [Ca2+](i). Furthermore glibenclamide did not reduce the oxotremorine- and baclofen-induced membrane hyperpolarization. In conclusion, the present results suggest that in striatal LA cells, O2/glucose deprivation activates a membrane hyperpolarization that does not involve ligand-gated K+ conductances but is sensitive to barium, glibenclamide, and charybdotoxin. The increase in [Ca2+](i) is partially due to influx through voltage-gated high-voltage- activated Ca2+ channels.

AB - The effects of combined O2/glucose deprivation were investigated on large aspiny (LA) interneurons recorded from a striatal slice preparation by means of simultaneous electrophysiological and optical recordings. LA interneurons were visually identified and impaled with sharp microelectrodes loaded with the calcium (Ca2+)-sensitive dye bis-fura-2. These cells showed the morphological, electrophysiological, and pharmacological features of large striatal cholinergic interneurons. O2/glucose deprivation induced a membrane hyperpolarization coupled to a concomitant increase in intracellular Ca2+ concentration ([Ca2+](i)). Interestingly, this [Ca2+](i) elevation was more pronounced in dendritic branches rather than in the somatic region. The O2/glucose-deprivation-induced membrane hyperpolarization reversed its polarity at the potassium (K+) equilibrium potential. Both membrane hyperpolarization and [Ca2+](i) rise were unaffected by TTX or by a combination of ionotropic glutamate receptors antagonists, D-2 amino-5- phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione. Sulfonylurea glibenclamide, a blocker of ATP-sensitive K+ channels, markedly reduced the O2/glucose-deprivation-induced membrane hyperpolarization but failed to prevent the rise in [Ca2+](i). Likewise, charybdotoxin, a large K+ channel (BK) inhibitor, abolished the membrane hyperpolarization but did not produce detectable changes of [Ca2+](i) elevation. A combination of high-voltage- activated Ca2+ channel blockers; significantly reduced both the membrane hyperpolarization and the rise in [Ca2+](i). In a set of experiments performed without dye in the recording electrode, either intracellular bis- (o-aminophenoxy)-N,N,N',N'-tetraacetic acid or external barium abolished the membrane hyperpolarization induced by O2/glucose deprivation. The hyperpolarizing effect on membrane potential was mimicked by oxotremorine, an M2-like muscarinic receptor agonist, and by baclofen, a GABA(B) receptor agonist. However, this membrane hyperpolarization was not coupled to an increase but rather to a decrease of the basal [Ca2+](i). Furthermore glibenclamide did not reduce the oxotremorine- and baclofen-induced membrane hyperpolarization. In conclusion, the present results suggest that in striatal LA cells, O2/glucose deprivation activates a membrane hyperpolarization that does not involve ligand-gated K+ conductances but is sensitive to barium, glibenclamide, and charybdotoxin. The increase in [Ca2+](i) is partially due to influx through voltage-gated high-voltage- activated Ca2+ channels.

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