Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures

Maira M. Fróes, Ana Helena P Correia, José Garcia-Abreu, David C. Spray, Antônio C. Campos De Carvalho, Vivaldo Moura Neto

Research output: Contribution to journalArticle

123 Citations (Scopus)

Abstract

Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic-nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.

Original languageEnglish (US)
Pages (from-to)7541-7546
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number13
DOIs
StatePublished - Jun 22 1999

Fingerprint

Astrocytes
Central Nervous System
Neurons
Coculture Techniques
nucleotidase
Coloring Agents
Brain
Communication
Octanols
Gap Junctions
Cyclic Nucleotides
Glial Fibrillary Acidic Protein
Oligodendroglia
Cerebrum
Tubulin
Fluorescent Dyes
Epitopes
Antibodies

Keywords

  • Double whole-cell voltage clamp
  • Glial-neuronal interactions
  • Lucifer yellow

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures. / Fróes, Maira M.; Correia, Ana Helena P; Garcia-Abreu, José; Spray, David C.; Campos De Carvalho, Antônio C.; Neto, Vivaldo Moura.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 13, 22.06.1999, p. 7541-7546.

Research output: Contribution to journalArticle

Fróes, Maira M. ; Correia, Ana Helena P ; Garcia-Abreu, José ; Spray, David C. ; Campos De Carvalho, Antônio C. ; Neto, Vivaldo Moura. / Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures. In: Proceedings of the National Academy of Sciences of the United States of America. 1999 ; Vol. 96, No. 13. pp. 7541-7546.
@article{c7819506c4294882aff700fd79dc9a4f,
title = "Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures",
abstract = "Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic-nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.",
keywords = "Double whole-cell voltage clamp, Glial-neuronal interactions, Lucifer yellow",
author = "Fr{\'o}es, {Maira M.} and Correia, {Ana Helena P} and Jos{\'e} Garcia-Abreu and Spray, {David C.} and {Campos De Carvalho}, {Ant{\^o}nio C.} and Neto, {Vivaldo Moura}",
year = "1999",
month = "6",
day = "22",
doi = "10.1073/pnas.96.13.7541",
language = "English (US)",
volume = "96",
pages = "7541--7546",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "13",

}

TY - JOUR

T1 - Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures

AU - Fróes, Maira M.

AU - Correia, Ana Helena P

AU - Garcia-Abreu, José

AU - Spray, David C.

AU - Campos De Carvalho, Antônio C.

AU - Neto, Vivaldo Moura

PY - 1999/6/22

Y1 - 1999/6/22

N2 - Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic-nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.

AB - Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against β-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic-nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.

KW - Double whole-cell voltage clamp

KW - Glial-neuronal interactions

KW - Lucifer yellow

UR - http://www.scopus.com/inward/record.url?scp=0033594984&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033594984&partnerID=8YFLogxK

U2 - 10.1073/pnas.96.13.7541

DO - 10.1073/pnas.96.13.7541

M3 - Article

C2 - 10377451

AN - SCOPUS:0033594984

VL - 96

SP - 7541

EP - 7546

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 13

ER -