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PMID: 11567043 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Global ischemia-induced increases in the gap junctional proteins connexin 32 (Cx32) and Cx36 in hippocampus and enhanced vulnerability of Cx32 knock-out mice.

Oguro K, Jover T, Tanaka H, Lin Y, Kojima T, Oguro N, Grooms SY, Bennett MV, Zukin RS

Abstract

Gap junctions are conductive channels that connect the interiors of coupled cells. In the hippocampus, GABA-containing hippocampal interneurons are interconnected by gap junctions, which mediate electrical coupling and synchronous firing and thereby promote inhibitory transmission. The present study was undertaken to examine the hypothesis that the gap junctional proteins connexin 32 (Cx32; expressed by oligodendrocytes, interneurons, or both), Cx36 (expressed by interneurons), and Cx43 (expressed by astrocytes) play a role in defining cell-specific patterns of neuronal death in hippocampus after global ischemia in mice. Global ischemia did not significantly alter Cx32 and Cx36 mRNA expression and slightly increased Cx43 mRNA expression in the vulnerable CA1, as assessed by Northern blot analysis and in situ hybridization. Global ischemia induced a selective increase in Cx32 and Cx36 but not Cx43 protein abundance in CA1 before onset of neuronal death, as assessed by Western blot analysis. The increase in Cx32 and Cx36 expression was intense and specific to parvalbumin-positive inhibitory interneurons of CA1, as assessed by double immunofluorescence. Protein abundance was unchanged in CA3 and dentate gyrus. The finding of increase in connexin protein without increase in mRNA suggests regulation of Cx32 and Cx36 expression at the translational or post-translational level. Cx32(Y/-) null mice exhibited enhanced vulnerability to brief ischemic insults, consistent with a role for Cx32 gap junctions in neuronal survival. These findings suggest that Cx32 and Cx36 gap junctions may contribute to the survival and resistance of GABAergic interneurons, thereby defining cell-specific patterns of global ischemia-induced neuronal death.

MeSH Terms
Animals Astrocytes/metabolism,pathology Brain Ischemia/genetics,metabolism,pathology Cell Survival/genetics Connexin 43/genetics,metabolism Connexins/deficiency,genetics,metabolism Down-Regulation Eye Proteins/genetics,metabolism Genetic Predisposition to Disease Hippocampus/metabolism,pathology Interneurons/metabolism,pathology Mice Mice, Knockout Neural Inhibition Oligodendroglia/metabolism,pathology Parvalbumins/biosynthesis RNA, Messenger/metabolism Up-Regulation
Chemicals
Connexin 43 Connexins Eye Proteins Parvalbumins RNA, Messenger connexin 32 connexin 36
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Oguro K
Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Jover T
Tanaka H
Lin Y
Kojima T
Oguro N
Grooms S Y
Bennett M V
Zukin R S
References (57)
57 references, click to expand
  1. The AMPAR subunit GluR2: still front and center-stage.
    Brain Res. 2000 Dec 15;886(1-2):190-207 PMID: 11119696
  2. Astrocytic gap junctional communication decreases neuronal vulnerability to oxidative stress-induced disruption of Ca2+ homeostasis and cell death.
    J Neurochem. 1998 Mar;70(3):958-70 PMID: 9489715
  3. Genetic determinants of susceptibility to excitotoxic cell death: implications for gene targeting approaches.
    Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):4103-8 PMID: 9108112
  4. Myelination defects and neuronal hyperexcitability in the neocortex of connexin 32-deficient mice.
    Cereb Cortex. 2000 Jul;10(7):684-97 PMID: 10906315
  5. Halothane reduces focal ischemic injury in the rat when brain temperature is controlled.
    Anesthesiology. 1995 May;82(5):1237-45; discussion 27A PMID: 7741299
  6. Gap junctions in cultured astrocytes: single-channel currents and characterization of channel-forming protein.
    Neuron. 1991 Jan;6(1):133-43 PMID: 1702648
  7. Immunorecognition, ultrastructure and phosphorylation status of astrocytic gap junctions and connexin43 in rat brain after cerebral focal ischaemia.
    Eur J Neurosci. 1998 Jul;10(7):2444-63 PMID: 9749772
  8. Vulnerability of mossy fiber targets in the rat hippocampus to forebrain ischemia.
    J Neurosci. 1993 Sep;13(9):3964-79 PMID: 8366355
  9. Differential expression of three gap junction proteins in developing and mature brain tissues.
    Proc Natl Acad Sci U S A. 1989 Dec;86(24):10148-52 PMID: 2557621
  10. Gap junctions linking the dendritic network of GABAergic interneurons in the hippocampus.
    J Neurosci. 2000 Feb 15;20(4):1519-28 PMID: 10662841
  11. Astrocyte glutamate transport: review of properties, regulation, and physiological functions.
    Glia. 2000 Oct;32(1):1-14 PMID: 10975906
  12. Gap junctions: new tools, new answers, new questions.
    Neuron. 1991 Mar;6(3):305-20 PMID: 1848077
  13. Synaptic distribution of the AMPA-GluR2 subunit and its colocalization with calcium-binding proteins in rat cerebral cortex: an immunohistochemical study using a GluR2-specific monoclonal antibody.
    Exp Neurol. 1996 Dec;142(2):296-312 PMID: 8934561
  14. Expression of Cx36 in mammalian neurons.
    Brain Res Brain Res Rev. 2000 Apr;32(1):72-85 PMID: 10751658
  15. Glia protect hippocampal neurons against excitatory amino acid-induced degeneration: involvement of fibroblast growth factor.
    Int J Dev Neurosci. 1990;8(4):399-415 PMID: 2251934
  16. Connexin expression in electrically coupled postnatal rat brain neurons.
    Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):10260-5 PMID: 10944183
  17. Anoxia blocks the presynaptic control of GABA release at inhibitory terminals in the rat hippocampus.
    Neuroscience. 1996 Dec;75(4):999-1002 PMID: 8938736
  18. Effective reduction of infarct volume by gap junction blockade in a rodent model of stroke.
    J Neurosurg. 1997 Dec;87(6):916-20 PMID: 9384404
  19. Selective vulnerability of the hippocampus in brain ischemia.
    Neuroscience. 1991;40(3):599-636 PMID: 1676492
  20. Structural abnormalities and deficient maintenance of peripheral nerve myelin in mice lacking the gap junction protein connexin 32.
    J Neurosci. 1997 Jun 15;17(12):4545-51 PMID: 9169515
  21. Astrocytes protect neurons from hydrogen peroxide toxicity.
    J Neurosci. 1996 Apr 15;16(8):2553-62 PMID: 8786431
  22. Global ischemia induces downregulation of Glur2 mRNA and increases AMPA receptor-mediated Ca2+ influx in hippocampal CA1 neurons of gerbil.
    J Neurosci. 1997 Aug 15;17(16):6179-88 PMID: 9236229
  23. Connections with connexins: the molecular basis of direct intercellular signaling.
    Eur J Biochem. 1996 May 15;238(1):1-27 PMID: 8665925
  24. Connexin32 in oligodendrocytes and association with myelinated fibers in mouse and rat brain.
    J Comp Neurol. 1997 Mar 24;379(4):571-91 PMID: 9067844
  25. The murine gap junction gene connexin36 is highly expressed in mouse retina and regulated during brain development.
    FEBS Lett. 1998 May 22;428(1-2):27-31 PMID: 9645468
  26. Temporal profile of neuronal damage in a model of transient forebrain ischemia.
    Ann Neurol. 1982 May;11(5):491-8 PMID: 7103425
  27. Delayed hippocampal damage in humans following cardiorespiratory arrest.
    Neurology. 1987 Aug;37(8):1281-6 PMID: 3614648
  28. Ischemia-induced cellular redistribution of the astrocytic gap junctional protein connexin43 in rat brain.
    Brain Res. 1994 Aug 1;652(2):311-22 PMID: 7953745
  29. Modulation of neuronal activity by glial cells in the retina.
    J Neurosci. 1998 Jun 1;18(11):4022-8 PMID: 9592083
  30. Reduction of infarct volume by halothane: effect on cerebral blood flow or perifocal spreading depression-like depolarizations.
    J Cereb Blood Flow Metab. 1997 Aug;17(8):857-64 PMID: 9290583
  31. From neuro-glue ('Nervenkitt') to glia: a prologue.
    Glia. 1998 Sep;24(1):1-7 PMID: 9700484
  32. Two networks of electrically coupled inhibitory neurons in neocortex.
    Nature. 1999 Nov 4;402(6757):75-9 PMID: 10573419
  33. Switch in glutamate receptor subunit gene expression in CA1 subfield of hippocampus following global ischemia in rats.
    Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10499-503 PMID: 1438239
  34. Connexin43: a protein from rat heart homologous to a gap junction protein from liver.
    J Cell Biol. 1987 Dec;105(6 Pt 1):2621-9 PMID: 2826492
  35. Cloning and expression of two related connexins from the perch retina define a distinct subgroup of the connexin family.
    J Neurosci. 1998 Oct 1;18(19):7625-37 PMID: 9742134
  36. Differential regulation of astrocyte TNF-alpha expression by the cytokines TGF-beta, IL-6 and IL-10.
    Int J Dev Neurosci. 1995 Jun-Jul;13(3-4):341-9 PMID: 7572286
  37. Disturbance of membrane function preceding ischemic delayed neuronal death in the gerbil hippocampus.
    J Cereb Blood Flow Metab. 1992 May;12(3):408-17 PMID: 1569136
  38. Fibroblast growth factors in the nervous system.
    J Neurobiol. 1994 Nov;25(11):1467-80 PMID: 7852998
  39. Rapid, small-scale RNA isolation from tissue culture cells.
    Biotechniques. 1991 Sep;11(3):324, 326-7 PMID: 1718328
  40. Gap-junction-mediated propagation and amplification of cell injury.
    Nat Neurosci. 1998 Oct;1(6):494-500 PMID: 10196547
  41. Coexpression of connexin45 and -32 in oligodendrocytes of rat brain.
    J Neurocytol. 1997 Jan;26(1):17-22 PMID: 9154525
  42. Knockdown of AMPA receptor GluR2 expression causes delayed neurodegeneration and increases damage by sublethal ischemia in hippocampal CA1 and CA3 neurons.
    J Neurosci. 1999 Nov 1;19(21):9218-27 PMID: 10531425
  43. Functional properties of channels formed by the neuronal gap junction protein connexin36.
    J Neurosci. 1999 Nov 15;19(22):9848-55 PMID: 10559394
  44. A network of fast-spiking cells in the neocortex connected by electrical synapses.
    Nature. 1999 Nov 4;402(6757):72-5 PMID: 10573418
  45. Connexin 35: a gap-junctional protein expressed preferentially in the skate retina.
    Mol Biol Cell. 1996 Feb;7(2):233-43 PMID: 8688555
  46. Delayed neuronal death in the gerbil hippocampus following ischemia.
    Brain Res. 1982 May 6;239(1):57-69 PMID: 7093691
  47. Remodeling of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor subunit composition in hippocampal neurons after global ischemia.
    Proc Natl Acad Sci U S A. 2000 Nov 21;97(24):13360-5 PMID: 11087875
  48. Electrically induced NGF production by astroglial cells.
    Nat Biotechnol. 1997 Feb;15(2):164-6 PMID: 9035143
  49. Connexins, connexons, and intercellular communication.
    Annu Rev Biochem. 1996;65:475-502 PMID: 8811187
  50. Defective propagation of signals generated by sympathetic nerve stimulation in the liver of connexin32-deficient mice.
    Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9565-70 PMID: 8790370
  51. A new model of bilateral hemispheric ischemia in the unanesthetized rat.
    Stroke. 1979 May-Jun;10(3):267-72 PMID: 37614
  52. Immunogold evidence that neuronal gap junctions in adult rat brain and spinal cord contain connexin-36 but not connexin-32 or connexin-43.
    Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7573-8 PMID: 10861019
  53. Cloning of a new gap junction gene (Cx36) highly expressed in mammalian brain neurons.
    Eur J Neurosci. 1998 Mar;10(3):1202-8 PMID: 9753189
  54. Receptor-mediated regulation of neuropeptide gene expression in astrocytes.
    Glia. 1994 Jun;11(2):185-90 PMID: 7927646
  55. Clustering of connexin 43-enhanced green fluorescent protein gap junction channels and functional coupling in living cells.
    Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2556-61 PMID: 10706639
  56. Molecular cloning of cDNA for rat liver gap junction protein.
    J Cell Biol. 1986 Jul;103(1):123-34 PMID: 3013898
  57. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2001-10-01
Pages
7534-42
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6762918
Subset
IM
Grants
NINDS NIH HHS · NS07412 · United States
NINDS NIH HHS · NS20752 · United States
NINDS NIH HHS · NS31282 · United States
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