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Major histocompatibility complex (MHC) class I gene expression in single neurons of the central nervous system: differential regulation by interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha.

Neumann H, Schmidt H, Cavalié A, Jenne D, Wekerle H - J. Exp. Med. (1997)

Bottom Line: The effect of tetrodotoxin is at least partly reverted by the neurotransmitter glutamate.In contrast to IFN-gamma, treatment with TNF-alpha did neither upregulate TAP1/TAP2 nor beta2-microglobulin gene expression, but induced MHC class I heavy chain gene transcription in all neurons.Consequently, no MHC class I molecules were detectable on the membranes of TNF-alpha-treated neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroimmunology, Max-Planck-Institute for Psychiatry, Martinsried, Germany.

ABSTRACT
This study examined the effect of the pro-inflammatory cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) on the induction of MHC class I-related genes in functionally mature brain neurons derived from cultures of dissociated rat hippocampal tissue. Patch clamp electrophysiology combined with single cell RT-PCR demonstrated that approximately 50% of the untreated neurons contained mRNA for MHC class I heavy chains, while, with few exceptions, the cells failed to transcribe beta2-microglobulin and TAP1/TAP2 gene transcripts. No constitutive expression of MHC class I protein was detectable by confocal laser microscopy on the surface of neurons. All neurons transcribed the alpha-chain of the interferon-type II receptor (binding IFN-gamma) along with the p55 receptor for TNF-alpha. Sustained exposure to IFN-gamma resulted in transcription of beta2-microglobulin and TAP1/TAP2 genes and MHC class I surface expression in a minor part of the neurons, but did not alter their electrophysiological activities as assessed by whole cell electrophysiology. Suppression of neuronal electric activity by the sodium channel blocker tetrodotoxin drastically increased to almost 100% IFN-gamma-mediated induction of MHC class I chains, of both TAP transporters, and of membrane expression of MHC class I protein. The effect of tetrodotoxin is at least partly reverted by the neurotransmitter glutamate. In contrast to IFN-gamma, treatment with TNF-alpha did neither upregulate TAP1/TAP2 nor beta2-microglobulin gene expression, but induced MHC class I heavy chain gene transcription in all neurons. Consequently, no MHC class I molecules were detectable on the membranes of TNF-alpha-treated neurons.

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MHC class I molecules  on the neuronal cell surface. Confocal laser scanning microscopy of  MHC class I molecules located on  the cell surface. Identification of individual neurons (pretreated with  IFN-γ for 72 h) by counterlabeling  the neuron-specific cytoskeleton  protein MAP2. MHC class I molecules labeled red (A) and MAP2  green (B). Merger of both images  from one optical section with specific labeling of MHC class I molecules on the neuronal cell surface  (C). D and E show one neuron  with, and another one without  MHC class I cell surface labeling.  Scale bar: (A–C) 10 μm; (D and E)  20 μm.
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Figure 8: MHC class I molecules on the neuronal cell surface. Confocal laser scanning microscopy of MHC class I molecules located on the cell surface. Identification of individual neurons (pretreated with IFN-γ for 72 h) by counterlabeling the neuron-specific cytoskeleton protein MAP2. MHC class I molecules labeled red (A) and MAP2 green (B). Merger of both images from one optical section with specific labeling of MHC class I molecules on the neuronal cell surface (C). D and E show one neuron with, and another one without MHC class I cell surface labeling. Scale bar: (A–C) 10 μm; (D and E) 20 μm.

Mentions: Membrane expression of MHC class I protein was studied by confocal laser scanning microscopy after double immunofluorescence labeling of live neurons combining monoclonal antibodies against MHC class I molecules with neuron specific marker antibodies. There was no constitutive expression of MHC class I molecules on the surface of untreated hippocampal neurons. Treatment with 100 U/ml IFN-γ for 72 h, however, resulted in MHC class I expression on the cell surface in 34% of the hippocampal neurons (Figs. 8 and 9 A). MHC class I molecules accumulated on the cell surface by IFN-γ steadily with the duration of treatment (Fig. 9 B).


Major histocompatibility complex (MHC) class I gene expression in single neurons of the central nervous system: differential regulation by interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha.

Neumann H, Schmidt H, Cavalié A, Jenne D, Wekerle H - J. Exp. Med. (1997)

MHC class I molecules  on the neuronal cell surface. Confocal laser scanning microscopy of  MHC class I molecules located on  the cell surface. Identification of individual neurons (pretreated with  IFN-γ for 72 h) by counterlabeling  the neuron-specific cytoskeleton  protein MAP2. MHC class I molecules labeled red (A) and MAP2  green (B). Merger of both images  from one optical section with specific labeling of MHC class I molecules on the neuronal cell surface  (C). D and E show one neuron  with, and another one without  MHC class I cell surface labeling.  Scale bar: (A–C) 10 μm; (D and E)  20 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196130&req=5

Figure 8: MHC class I molecules on the neuronal cell surface. Confocal laser scanning microscopy of MHC class I molecules located on the cell surface. Identification of individual neurons (pretreated with IFN-γ for 72 h) by counterlabeling the neuron-specific cytoskeleton protein MAP2. MHC class I molecules labeled red (A) and MAP2 green (B). Merger of both images from one optical section with specific labeling of MHC class I molecules on the neuronal cell surface (C). D and E show one neuron with, and another one without MHC class I cell surface labeling. Scale bar: (A–C) 10 μm; (D and E) 20 μm.
Mentions: Membrane expression of MHC class I protein was studied by confocal laser scanning microscopy after double immunofluorescence labeling of live neurons combining monoclonal antibodies against MHC class I molecules with neuron specific marker antibodies. There was no constitutive expression of MHC class I molecules on the surface of untreated hippocampal neurons. Treatment with 100 U/ml IFN-γ for 72 h, however, resulted in MHC class I expression on the cell surface in 34% of the hippocampal neurons (Figs. 8 and 9 A). MHC class I molecules accumulated on the cell surface by IFN-γ steadily with the duration of treatment (Fig. 9 B).

Bottom Line: The effect of tetrodotoxin is at least partly reverted by the neurotransmitter glutamate.In contrast to IFN-gamma, treatment with TNF-alpha did neither upregulate TAP1/TAP2 nor beta2-microglobulin gene expression, but induced MHC class I heavy chain gene transcription in all neurons.Consequently, no MHC class I molecules were detectable on the membranes of TNF-alpha-treated neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroimmunology, Max-Planck-Institute for Psychiatry, Martinsried, Germany.

ABSTRACT
This study examined the effect of the pro-inflammatory cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) on the induction of MHC class I-related genes in functionally mature brain neurons derived from cultures of dissociated rat hippocampal tissue. Patch clamp electrophysiology combined with single cell RT-PCR demonstrated that approximately 50% of the untreated neurons contained mRNA for MHC class I heavy chains, while, with few exceptions, the cells failed to transcribe beta2-microglobulin and TAP1/TAP2 gene transcripts. No constitutive expression of MHC class I protein was detectable by confocal laser microscopy on the surface of neurons. All neurons transcribed the alpha-chain of the interferon-type II receptor (binding IFN-gamma) along with the p55 receptor for TNF-alpha. Sustained exposure to IFN-gamma resulted in transcription of beta2-microglobulin and TAP1/TAP2 genes and MHC class I surface expression in a minor part of the neurons, but did not alter their electrophysiological activities as assessed by whole cell electrophysiology. Suppression of neuronal electric activity by the sodium channel blocker tetrodotoxin drastically increased to almost 100% IFN-gamma-mediated induction of MHC class I chains, of both TAP transporters, and of membrane expression of MHC class I protein. The effect of tetrodotoxin is at least partly reverted by the neurotransmitter glutamate. In contrast to IFN-gamma, treatment with TNF-alpha did neither upregulate TAP1/TAP2 nor beta2-microglobulin gene expression, but induced MHC class I heavy chain gene transcription in all neurons. Consequently, no MHC class I molecules were detectable on the membranes of TNF-alpha-treated neurons.

Show MeSH
Related in: MedlinePlus