Hiscock J J, MacKenzie L, Willoughby J O
Centre for Neuroscience, Flinders University and Medical Centre, Adelaide, Australia.
Brain Res. 1996 Nov 4;738(2):301-12. doi: 10.1016/s0006-8993(96)00806-2.
In adult rats single seizures of varying behavioural severities were caused by slow, systemic infusion of picrotoxin, an antagonist of the C1- channel at the GABAA receptor. We used a double labelling immunohistochemical method to define the subclasses of neurons that contained Fos protein following seizures. In four cortical regions (piriform, entorhinal, motor and sensory) neuronal subclasses were defined with antibodies against the calcium-binding proteins D-28K, parvalbumin and calretinin (aspiny neurons), and neurofilament protein (spiny neurons). The remaining spiny neuron population was estimated by subtraction of defined subclasses from total neuronal numbers determined from Nissl stain. After seizures, most of the calbindin D-28K immunoreactive interneurons (> 80%) and many of the unlabelled spiny neurons (60-80%) were FOs positive. Co-localisation of Fos was found in about 30% of parvalbumin, calretinin and neurofilament protein immunoreactive neurons. Paradoxically, mild seizures were associated with induction of Fos in up to 80% of cortical cells and more severe seizures with 60%, the difference being due to different levels of Fos induction in spiny neurons. These results also demonstrate that seizures induce Fos predominantly in excitatory cortical neurons.
在成年大鼠中,通过缓慢、全身性输注印防己毒素(一种GABAA受体氯离子通道拮抗剂)引发了行为严重程度各异的单次癫痫发作。我们采用双重标记免疫组织化学方法来确定癫痫发作后含有Fos蛋白的神经元亚类。在四个皮质区域(梨状皮质、内嗅皮质、运动皮质和感觉皮质),使用针对钙结合蛋白D-28K、小白蛋白和钙视网膜蛋白(无棘神经元)以及神经丝蛋白(有棘神经元)的抗体来定义神经元亚类。通过从尼氏染色确定的总神经元数量中减去已定义的亚类来估计其余有棘神经元群体。癫痫发作后,大多数钙结合蛋白D-28K免疫反应性中间神经元(>80%)和许多未标记的有棘神经元(60-80%)Fos呈阳性。在约30%的小白蛋白、钙视网膜蛋白和神经丝蛋白免疫反应性神经元中发现了Fos的共定位。矛盾的是,轻度癫痫发作与高达80%的皮质细胞中Fos的诱导有关,而重度癫痫发作与60%的皮质细胞中Fos的诱导有关,差异源于有棘神经元中Fos诱导水平的不同。这些结果还表明,癫痫发作主要在兴奋性皮质神经元中诱导Fos。