The role of sulfhydryl groups in d-aspartate and rubidium release from neonatal rat primary astrocyte cultures

Michael Aschner, K. J. Mullaney, M. N. Fehm, D. Vitarella, D. E. Wagoner, H. K. Kimelberg

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have recently demonstrated that both methylmercury (MeHg) and mercuric chloride (MC) induce d-aspartate release from neonatal rat primary astrocyte cultures maintained in isotonic conditions [1,3,21,22]. In the present study, we compare several other sulfhydryl-(-SH) selective alkylating reagents [methyl methanethiosulfonate (MMTS), N-ehtylmaleimide (NEM), and iodoacetamide (IA)] in isotonic, as well as hypotonic conditions to discern the functional importance of -SH groups in [3H]d-aspartate and 86rubidium (86Rb) release from astrocytes. Treatment of astrocytes (5 min) in isotonic buffer with the hydrophobic reagent NEM (10 μM) caused a marked increase in 86Rb release but had no effect of [3H]d-aspartate release. Neither IA-, nor MMTS-treatment (both at 10 μM) induced increase in [3H]d-aspartate of 86Rb release in isotonic buffer. In hypotonic condition (-50 mM Na+), astrocytes were most sensitive to MC exposure (5 μM), exhibiting an increase in both [3H]d-aspartate and 86Rb efflux. The hydrophobic compounds MMTS and NEM, and the hydrophilic -SH modifying reagent, IA, attenuated the hypotonic-induced efflux of [3H]d-aspartate, in the absence of an effect of 86Rb release. These observations are consistent with a critical role for -SH groups both in basal (i.e. isotonic) and hypotonic-induced release of d-aspartate and Rb from astrocytes. Lack of uniformity of these effects may be attributed to site-specificity, related to the physicochemical properties of these -SH alkylating reagents.

Original languageEnglish (US)
Pages (from-to)16-23
Number of pages8
JournalBrain Research
Volume648
Issue number1
DOIs
StatePublished - Jun 13 1994
Externally publishedYes

Fingerprint

Rubidium
Aspartic Acid
Astrocytes
Iodoacetamide
Sulfhydryl Reagents
Mercuric Chloride
Buffers

Keywords

  • -SH group
  • Astrocyte
  • d-Aspartate
  • Hypotonicity
  • Regulatory volume decrease
  • Rubidium

ASJC Scopus subject areas

  • Developmental Biology
  • Molecular Biology
  • Clinical Neurology
  • Neuroscience(all)

Cite this

The role of sulfhydryl groups in d-aspartate and rubidium release from neonatal rat primary astrocyte cultures. / Aschner, Michael; Mullaney, K. J.; Fehm, M. N.; Vitarella, D.; Wagoner, D. E.; Kimelberg, H. K.

In: Brain Research, Vol. 648, No. 1, 13.06.1994, p. 16-23.

Research output: Contribution to journalArticle

Aschner, Michael ; Mullaney, K. J. ; Fehm, M. N. ; Vitarella, D. ; Wagoner, D. E. ; Kimelberg, H. K. / The role of sulfhydryl groups in d-aspartate and rubidium release from neonatal rat primary astrocyte cultures. In: Brain Research. 1994 ; Vol. 648, No. 1. pp. 16-23.
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AU - Aschner, Michael

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AB - We have recently demonstrated that both methylmercury (MeHg) and mercuric chloride (MC) induce d-aspartate release from neonatal rat primary astrocyte cultures maintained in isotonic conditions [1,3,21,22]. In the present study, we compare several other sulfhydryl-(-SH) selective alkylating reagents [methyl methanethiosulfonate (MMTS), N-ehtylmaleimide (NEM), and iodoacetamide (IA)] in isotonic, as well as hypotonic conditions to discern the functional importance of -SH groups in [3H]d-aspartate and 86rubidium (86Rb) release from astrocytes. Treatment of astrocytes (5 min) in isotonic buffer with the hydrophobic reagent NEM (10 μM) caused a marked increase in 86Rb release but had no effect of [3H]d-aspartate release. Neither IA-, nor MMTS-treatment (both at 10 μM) induced increase in [3H]d-aspartate of 86Rb release in isotonic buffer. In hypotonic condition (-50 mM Na+), astrocytes were most sensitive to MC exposure (5 μM), exhibiting an increase in both [3H]d-aspartate and 86Rb efflux. The hydrophobic compounds MMTS and NEM, and the hydrophilic -SH modifying reagent, IA, attenuated the hypotonic-induced efflux of [3H]d-aspartate, in the absence of an effect of 86Rb release. These observations are consistent with a critical role for -SH groups both in basal (i.e. isotonic) and hypotonic-induced release of d-aspartate and Rb from astrocytes. Lack of uniformity of these effects may be attributed to site-specificity, related to the physicochemical properties of these -SH alkylating reagents.

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