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Interferon gamma gene expression in sensory neurons: evidence for autocrine gene regulation.

Neumann H, Schmidt H, Wilharm E, Behrens L, Wekerle H - J. Exp. Med. (1997)

Bottom Line: Locally produced IFN-gamma acts back on its cellular source.Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-gamma-dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures.Our findings indicate a role of IFN-gamma in autocrine regulation of sensory neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroimmunology, Max-Planck-Institute for Psychiatry, D-82152 Martinsried, Germany.

ABSTRACT
We explored expression and possible function of interferon-gamma (IFN-gamma) in cultured fetal (E15) rat dorsal root ganglion neurons combining whole cell patch-clamp electrophysiology with single cell reverse transcriptase polymerase chain reaction and confocal laser immunocytochemistry. Morphologically, we located IFN-gamma protein in the cytoplasm of the neurons in culture as well as in situ during peri- and postnatal development. Transcripts for classic IFN-gamma and for its receptor were determined in probes of cytoplasm sampled from individual cultured neurons, which had been identified by patch clamp electrophysiology. In addition, the cultured neurons expressed both chains of the IFN-gamma receptor. Locally produced IFN-gamma acts back on its cellular source. Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-gamma-dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures. However, both processes were substantially blocked in the presence of antibodies neutralizing IFN-gamma. Our findings indicate a role of IFN-gamma in autocrine regulation of sensory neurons.

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IFN-γ immunoreactivity in cultured sensory neurons. (A) IFN-γ immunofluorescence labeling in neurons.  Embryonic sensory neurons cultured for 7 d were identified by  antibodies recognizing neurofilament and were double-labeled  with antibodies directed against  IFN-γ. Scale bar: 10 μm. (B)  Confocal laser scanning microscopy of sensory neurons. Neurofilament labeling is located in  the cytoplasm and the neuronal  processes, whereas IFN-γ labeling is only localized in the perinuclear cytoplasm. Scale bar: 10  μm. (C) Incubation with primary control antibodies instead  of IFN-γ–specific antibodies  showed baseline labeling intensity. Scale bar: 10 μm.
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Figure 2: IFN-γ immunoreactivity in cultured sensory neurons. (A) IFN-γ immunofluorescence labeling in neurons. Embryonic sensory neurons cultured for 7 d were identified by antibodies recognizing neurofilament and were double-labeled with antibodies directed against IFN-γ. Scale bar: 10 μm. (B) Confocal laser scanning microscopy of sensory neurons. Neurofilament labeling is located in the cytoplasm and the neuronal processes, whereas IFN-γ labeling is only localized in the perinuclear cytoplasm. Scale bar: 10 μm. (C) Incubation with primary control antibodies instead of IFN-γ–specific antibodies showed baseline labeling intensity. Scale bar: 10 μm.

Mentions: DRG cells from embryonic rats (E15) were cultured in the presence of nerve growth factor. Most of the DRG neurons that differentiated in vitro bound monoclonal antibodies recognizing IFN-γ. Specific immunofluorescence was detected after 7 d in culture in 83 ± 8% of DRG neurons identified by neurofilament labeling, but in none of the nonneuronal cells (Fig. 2). Confocal laser scanning microscopy localized the IFN-γ proteins in the cytoplasm, sparing the nucleus and the cell membrane (Fig. 2), whereas processes and dendrites showed no or only very weak reactivity.


Interferon gamma gene expression in sensory neurons: evidence for autocrine gene regulation.

Neumann H, Schmidt H, Wilharm E, Behrens L, Wekerle H - J. Exp. Med. (1997)

IFN-γ immunoreactivity in cultured sensory neurons. (A) IFN-γ immunofluorescence labeling in neurons.  Embryonic sensory neurons cultured for 7 d were identified by  antibodies recognizing neurofilament and were double-labeled  with antibodies directed against  IFN-γ. Scale bar: 10 μm. (B)  Confocal laser scanning microscopy of sensory neurons. Neurofilament labeling is located in  the cytoplasm and the neuronal  processes, whereas IFN-γ labeling is only localized in the perinuclear cytoplasm. Scale bar: 10  μm. (C) Incubation with primary control antibodies instead  of IFN-γ–specific antibodies  showed baseline labeling intensity. Scale bar: 10 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199162&req=5

Figure 2: IFN-γ immunoreactivity in cultured sensory neurons. (A) IFN-γ immunofluorescence labeling in neurons. Embryonic sensory neurons cultured for 7 d were identified by antibodies recognizing neurofilament and were double-labeled with antibodies directed against IFN-γ. Scale bar: 10 μm. (B) Confocal laser scanning microscopy of sensory neurons. Neurofilament labeling is located in the cytoplasm and the neuronal processes, whereas IFN-γ labeling is only localized in the perinuclear cytoplasm. Scale bar: 10 μm. (C) Incubation with primary control antibodies instead of IFN-γ–specific antibodies showed baseline labeling intensity. Scale bar: 10 μm.
Mentions: DRG cells from embryonic rats (E15) were cultured in the presence of nerve growth factor. Most of the DRG neurons that differentiated in vitro bound monoclonal antibodies recognizing IFN-γ. Specific immunofluorescence was detected after 7 d in culture in 83 ± 8% of DRG neurons identified by neurofilament labeling, but in none of the nonneuronal cells (Fig. 2). Confocal laser scanning microscopy localized the IFN-γ proteins in the cytoplasm, sparing the nucleus and the cell membrane (Fig. 2), whereas processes and dendrites showed no or only very weak reactivity.

Bottom Line: Locally produced IFN-gamma acts back on its cellular source.Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-gamma-dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures.Our findings indicate a role of IFN-gamma in autocrine regulation of sensory neurons.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroimmunology, Max-Planck-Institute for Psychiatry, D-82152 Martinsried, Germany.

ABSTRACT
We explored expression and possible function of interferon-gamma (IFN-gamma) in cultured fetal (E15) rat dorsal root ganglion neurons combining whole cell patch-clamp electrophysiology with single cell reverse transcriptase polymerase chain reaction and confocal laser immunocytochemistry. Morphologically, we located IFN-gamma protein in the cytoplasm of the neurons in culture as well as in situ during peri- and postnatal development. Transcripts for classic IFN-gamma and for its receptor were determined in probes of cytoplasm sampled from individual cultured neurons, which had been identified by patch clamp electrophysiology. In addition, the cultured neurons expressed both chains of the IFN-gamma receptor. Locally produced IFN-gamma acts back on its cellular source. Phosphorylation and nuclear translocation of the IFN-inducible transcriptional factor STAT1 as well as IFN-gamma-dependent expression of major histocompatibility complex class I molecules on the neuronal membrane were noted in untreated cultures. However, both processes were substantially blocked in the presence of antibodies neutralizing IFN-gamma. Our findings indicate a role of IFN-gamma in autocrine regulation of sensory neurons.

Show MeSH
Related in: MedlinePlus