Neumann H, Schmidt H, Wilharm E, Behrens L, Wekerle H
Department of Neuroimmunology, Max-Planck-Institute for Psychiatry, D-82152 Martinsried, Germany.
J Exp Med. 1997 Dec 15;186(12):2023-31. doi: 10.1084/jem.186.12.2023.
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.
我们将全细胞膜片钳电生理学与单细胞逆转录聚合酶链反应及共聚焦激光免疫细胞化学相结合，探讨了干扰素-γ（IFN-γ）在培养的胎鼠（E15）背根神经节神经元中的表达及可能的功能。在形态学上，我们在培养的神经元细胞质中以及围产期和出生后发育过程中的原位发现了IFN-γ蛋白。从经膜片钳电生理学鉴定的单个培养神经元中采集的细胞质探针中，检测到了经典IFN-γ及其受体的转录本。此外，培养的神经元表达了IFN-γ受体的两条链。局部产生的IFN-γ会作用于其细胞来源。在未经处理的培养物中，观察到了IFN诱导的转录因子STAT1的磷酸化和核转位，以及神经元膜上主要组织相容性复合体I类分子的IFN-γ依赖性表达。然而，在存在中和IFN-γ的抗体时，这两个过程均被显著阻断。我们的研究结果表明IFN-γ在感觉神经元的自分泌调节中发挥作用。
