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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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Identification of neurons by whole cell patch-clamp electrophysiology. (A) Action potentials were evoked in sensory neurons cultured for 7 d by current pulses of increasing amplitude (200, 300, and 400  pA) and were recorded in the current clamp mode. (B) Whole cell membrane currents were evoked by successive depolarization steps of 10 mV  (from a holding potential of −80 mV to voltages ranging from −50 to  +50 mV) and were recorded in the voltage clamp mode.
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Figure 3: Identification of neurons by whole cell patch-clamp electrophysiology. (A) Action potentials were evoked in sensory neurons cultured for 7 d by current pulses of increasing amplitude (200, 300, and 400 pA) and were recorded in the current clamp mode. (B) Whole cell membrane currents were evoked by successive depolarization steps of 10 mV (from a holding potential of −80 mV to voltages ranging from −50 to +50 mV) and were recorded in the voltage clamp mode.

Mentions: After 7 d in culture, neurons could be distinguished morphologically from nonneuronal cells. Their neuronal identity was ascertained by whole cell patch-clamp electrophysiology (15). As expected, the DRG-derived neurons showed sodium currents activated at different membrane potentials, and responded to current pulses of increasing amplitude with sodium action potentials (Fig. 3).


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)

Identification of neurons by whole cell patch-clamp electrophysiology. (A) Action potentials were evoked in sensory neurons cultured for 7 d by current pulses of increasing amplitude (200, 300, and 400  pA) and were recorded in the current clamp mode. (B) Whole cell membrane currents were evoked by successive depolarization steps of 10 mV  (from a holding potential of −80 mV to voltages ranging from −50 to  +50 mV) and were recorded in the voltage clamp mode.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 3: Identification of neurons by whole cell patch-clamp electrophysiology. (A) Action potentials were evoked in sensory neurons cultured for 7 d by current pulses of increasing amplitude (200, 300, and 400 pA) and were recorded in the current clamp mode. (B) Whole cell membrane currents were evoked by successive depolarization steps of 10 mV (from a holding potential of −80 mV to voltages ranging from −50 to +50 mV) and were recorded in the voltage clamp mode.
Mentions: After 7 d in culture, neurons could be distinguished morphologically from nonneuronal cells. Their neuronal identity was ascertained by whole cell patch-clamp electrophysiology (15). As expected, the DRG-derived neurons showed sodium currents activated at different membrane potentials, and responded to current pulses of increasing amplitude with sodium action potentials (Fig. 3).

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