Effects of volatile anesthetics on glutamate transporter, excitatory amino acid transporter type 3: The role of protein kinase C

Sanghwan Do, Ganesan L. Kamatchi, Jacqueline M. Washington, Zhiyi Zuo

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Background: Glutamate transporters play an important role in maintaining extracellular glutamate homeostasis. The authors studied the effects of volatile anesthetics on one type of glutamate transporters, excitatory amino acid transporter type 3 (EAAT3), and the role of protein kinase C in mediating these effects. Methods: Excitatory amino acid transporter type 3 was expressed inXenopus oocytes by injection of EAAT3 mRNA. Using two-electrode voltage clamp, membrane currents were recorded before, during, and after application of L-glutamate. Responses were quantified by integrating the current trace and are reported as microcoulombs. Data are mean ± SEM. Results: L-Glutamate-induced responses were increased gradually with the increased concentrations of isoflurane, a volatile anesthetic. At 0.52 and 0.70 mM isoflurane, the inward current was significantly increased compared with control. Isoflurane (0.70 mM) significantly increased Vmax (maximum velocity) (3.6 ± 0.4 to 5.1 ± 0.4 μC; P < 0.05) but not Km (Michoelis-Menten Constant) (55.4 ± 17.0 vs. 61.7 ± 13.6 μM; P > 0.05) of EAAT3 for glutamate compared with control. Treatment of the oocytes with phorbol-12-myrisate-13-acetate, a protein kinase C activator, caused a significant increase in transporter current (1.7 ± 0.2 to 2.5 ± 0.2 μC; P < 0.05). Responses in the presence of the combination of phorbol-12-myrisate-13-acetate and volatile anesthetics (isoflurane, halothane, or sevoflurane) were not greater than those when volatile anesthetic was present alone. Oocytes pretreated with any of the three protein kinase C inhibitors alone (chelerythrine, staurosporine, or calphostin C) did not affect basal transporter current. Although chelerythrine did not change the anesthetic effects on the activity of EAAT3, staurosporine or calphostin C abolished the anesthetic-induced increase of EAAT3 activity. Conclusions: These data suggest that volatile anesthetics enhance EAAT3 activity and that protein kinase C is involved in mediating these anesthetic effects.

Original languageEnglish
Pages (from-to)1492-1497
Number of pages6
JournalAnesthesiology
Volume96
Issue number6
DOIs
StatePublished - 13 Jun 2002

Fingerprint

Excitatory Amino Acid Transporter 3
Amino Acid Transport System X-AG
Protein Kinase C
Anesthetics
Isoflurane
Glutamic Acid
Oocytes
Staurosporine
Acetates
Protein C Inhibitor
Halothane
Protein Kinase Inhibitors
Electrodes
Homeostasis

Cite this

Do, Sanghwan ; Kamatchi, Ganesan L. ; Washington, Jacqueline M. ; Zuo, Zhiyi. / Effects of volatile anesthetics on glutamate transporter, excitatory amino acid transporter type 3 : The role of protein kinase C. In: Anesthesiology. 2002 ; Vol. 96, No. 6. pp. 1492-1497.
@article{f5ca8f9ead1c41338b91a464f6b8f1f5,
title = "Effects of volatile anesthetics on glutamate transporter, excitatory amino acid transporter type 3: The role of protein kinase C",
abstract = "Background: Glutamate transporters play an important role in maintaining extracellular glutamate homeostasis. The authors studied the effects of volatile anesthetics on one type of glutamate transporters, excitatory amino acid transporter type 3 (EAAT3), and the role of protein kinase C in mediating these effects. Methods: Excitatory amino acid transporter type 3 was expressed inXenopus oocytes by injection of EAAT3 mRNA. Using two-electrode voltage clamp, membrane currents were recorded before, during, and after application of L-glutamate. Responses were quantified by integrating the current trace and are reported as microcoulombs. Data are mean ± SEM. Results: L-Glutamate-induced responses were increased gradually with the increased concentrations of isoflurane, a volatile anesthetic. At 0.52 and 0.70 mM isoflurane, the inward current was significantly increased compared with control. Isoflurane (0.70 mM) significantly increased Vmax (maximum velocity) (3.6 ± 0.4 to 5.1 ± 0.4 μC; P < 0.05) but not Km (Michoelis-Menten Constant) (55.4 ± 17.0 vs. 61.7 ± 13.6 μM; P > 0.05) of EAAT3 for glutamate compared with control. Treatment of the oocytes with phorbol-12-myrisate-13-acetate, a protein kinase C activator, caused a significant increase in transporter current (1.7 ± 0.2 to 2.5 ± 0.2 μC; P < 0.05). Responses in the presence of the combination of phorbol-12-myrisate-13-acetate and volatile anesthetics (isoflurane, halothane, or sevoflurane) were not greater than those when volatile anesthetic was present alone. Oocytes pretreated with any of the three protein kinase C inhibitors alone (chelerythrine, staurosporine, or calphostin C) did not affect basal transporter current. Although chelerythrine did not change the anesthetic effects on the activity of EAAT3, staurosporine or calphostin C abolished the anesthetic-induced increase of EAAT3 activity. Conclusions: These data suggest that volatile anesthetics enhance EAAT3 activity and that protein kinase C is involved in mediating these anesthetic effects.",
author = "Sanghwan Do and Kamatchi, {Ganesan L.} and Washington, {Jacqueline M.} and Zhiyi Zuo",
year = "2002",
month = "6",
day = "13",
doi = "10.1097/00000542-200206000-00032",
language = "English",
volume = "96",
pages = "1492--1497",
journal = "Anesthesiology",
issn = "0003-3022",
publisher = "Lippincott Williams and Wilkins Ltd.",
number = "6",

}

Effects of volatile anesthetics on glutamate transporter, excitatory amino acid transporter type 3 : The role of protein kinase C. / Do, Sanghwan; Kamatchi, Ganesan L.; Washington, Jacqueline M.; Zuo, Zhiyi.

In: Anesthesiology, Vol. 96, No. 6, 13.06.2002, p. 1492-1497.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effects of volatile anesthetics on glutamate transporter, excitatory amino acid transporter type 3

T2 - The role of protein kinase C

AU - Do, Sanghwan

AU - Kamatchi, Ganesan L.

AU - Washington, Jacqueline M.

AU - Zuo, Zhiyi

PY - 2002/6/13

Y1 - 2002/6/13

N2 - Background: Glutamate transporters play an important role in maintaining extracellular glutamate homeostasis. The authors studied the effects of volatile anesthetics on one type of glutamate transporters, excitatory amino acid transporter type 3 (EAAT3), and the role of protein kinase C in mediating these effects. Methods: Excitatory amino acid transporter type 3 was expressed inXenopus oocytes by injection of EAAT3 mRNA. Using two-electrode voltage clamp, membrane currents were recorded before, during, and after application of L-glutamate. Responses were quantified by integrating the current trace and are reported as microcoulombs. Data are mean ± SEM. Results: L-Glutamate-induced responses were increased gradually with the increased concentrations of isoflurane, a volatile anesthetic. At 0.52 and 0.70 mM isoflurane, the inward current was significantly increased compared with control. Isoflurane (0.70 mM) significantly increased Vmax (maximum velocity) (3.6 ± 0.4 to 5.1 ± 0.4 μC; P < 0.05) but not Km (Michoelis-Menten Constant) (55.4 ± 17.0 vs. 61.7 ± 13.6 μM; P > 0.05) of EAAT3 for glutamate compared with control. Treatment of the oocytes with phorbol-12-myrisate-13-acetate, a protein kinase C activator, caused a significant increase in transporter current (1.7 ± 0.2 to 2.5 ± 0.2 μC; P < 0.05). Responses in the presence of the combination of phorbol-12-myrisate-13-acetate and volatile anesthetics (isoflurane, halothane, or sevoflurane) were not greater than those when volatile anesthetic was present alone. Oocytes pretreated with any of the three protein kinase C inhibitors alone (chelerythrine, staurosporine, or calphostin C) did not affect basal transporter current. Although chelerythrine did not change the anesthetic effects on the activity of EAAT3, staurosporine or calphostin C abolished the anesthetic-induced increase of EAAT3 activity. Conclusions: These data suggest that volatile anesthetics enhance EAAT3 activity and that protein kinase C is involved in mediating these anesthetic effects.

AB - Background: Glutamate transporters play an important role in maintaining extracellular glutamate homeostasis. The authors studied the effects of volatile anesthetics on one type of glutamate transporters, excitatory amino acid transporter type 3 (EAAT3), and the role of protein kinase C in mediating these effects. Methods: Excitatory amino acid transporter type 3 was expressed inXenopus oocytes by injection of EAAT3 mRNA. Using two-electrode voltage clamp, membrane currents were recorded before, during, and after application of L-glutamate. Responses were quantified by integrating the current trace and are reported as microcoulombs. Data are mean ± SEM. Results: L-Glutamate-induced responses were increased gradually with the increased concentrations of isoflurane, a volatile anesthetic. At 0.52 and 0.70 mM isoflurane, the inward current was significantly increased compared with control. Isoflurane (0.70 mM) significantly increased Vmax (maximum velocity) (3.6 ± 0.4 to 5.1 ± 0.4 μC; P < 0.05) but not Km (Michoelis-Menten Constant) (55.4 ± 17.0 vs. 61.7 ± 13.6 μM; P > 0.05) of EAAT3 for glutamate compared with control. Treatment of the oocytes with phorbol-12-myrisate-13-acetate, a protein kinase C activator, caused a significant increase in transporter current (1.7 ± 0.2 to 2.5 ± 0.2 μC; P < 0.05). Responses in the presence of the combination of phorbol-12-myrisate-13-acetate and volatile anesthetics (isoflurane, halothane, or sevoflurane) were not greater than those when volatile anesthetic was present alone. Oocytes pretreated with any of the three protein kinase C inhibitors alone (chelerythrine, staurosporine, or calphostin C) did not affect basal transporter current. Although chelerythrine did not change the anesthetic effects on the activity of EAAT3, staurosporine or calphostin C abolished the anesthetic-induced increase of EAAT3 activity. Conclusions: These data suggest that volatile anesthetics enhance EAAT3 activity and that protein kinase C is involved in mediating these anesthetic effects.

UR - http://www.scopus.com/inward/record.url?scp=0036260450&partnerID=8YFLogxK

U2 - 10.1097/00000542-200206000-00032

DO - 10.1097/00000542-200206000-00032

M3 - Article

C2 - 12170065

AN - SCOPUS:0036260450

VL - 96

SP - 1492

EP - 1497

JO - Anesthesiology

JF - Anesthesiology

SN - 0003-3022

IS - 6

ER -