Establishment of analyzing method to delineate the central nervous system active in rat with treadmill running and its application to exercise physiology.

我们的目标是描绘在速度@乳酸阈值(LT)下运行的大鼠大脑中的活动部位。因此，我们使用我们的跑步模型，即雄性大鼠在跑步机上跑步。在LT运行过程中和运行后，取出整个大脑并切取额叶进行c-fos的原位杂交(Exp1)。此外，我们还进一步研究了在运动阈值为*20的条件下，电刺激坐骨神经引起肌肉收缩后，大脑如何表达c-fos。实验采用免疫组织化学方法检测c-fos蛋白表达。在实验中，我们还应检测c-fos表达细胞中精氨酸加压素(AVP)的免疫反应性。在实验一中，c-fos基因在下丘脑(室旁核和视上核)、丘脑、海马区和边缘区以及运动中枢和顶区等部位均有表达。实验二，AVP细胞c-fos阳性表达，提示AVP神经元可能参与了跑步应激。只有在“运动压力反应”的情况下，才能观察到c-fos蛋白在表达AVP的神经细胞中的表达。这些结果表明，大鼠大脑中的几个基因位点在LT运行过程中可能是活跃的。尤其是下丘脑AVP神经元在跑步过程中会出现反射性兴奋。这可能表明中等强度的跑步运动可以刺激中枢神经系统的几个区域，尤其是下丘脑。最后，我们试图通过c-fos的原位杂交来研究支配比目鱼肌(MN-SOL)和足底(MN-PL)的MN如何在LT跑步中被招募。虽然MN-SOL中c-fos的表达程度没有变化，但MN-PL的表达略高于LT运动，提示这些检测c-fos基因/蛋白的方法可用于探索跑步大鼠中枢神经系统的活性部位。

We aimed to delineate the active site in rat brain running at the speed @ lactate threshold (LT). We therefore use our running model in which in male rat on the treadmill. During and after the LT running, whole brain was removed and sectioned frontal for an in situ hybridization for c-fos (Exp.1). Moreover, we further examined how the brain could express the c-fos after muscle contraction induced by an electrical atimulation of the sciatic nerve at the condition with *20 motorthreshold. In the expreiment, we examined c-fos protein by an immunohistochemistry. In the experiment, we should also examined the immunoreactivity of arginine-vasopressin (AVP) in the c-fos expressing cells. In the Exp.I,the c-fos mRNA was shown to be increasedin the loci such as hypothalamus (paraventricular nucleus and supraoptic nucleus) thalamus, hippocampus, and limbic area as well as motor center as parietal region. In the Exp.II,the positive staining with c-fos was found in the AVP cell, suggesting that AVP neuron will be involved in the running stress. The c-fos protein expressed within the neuronal cells expressing AVP was only observed in the condition that "exercise pressure response" was occurred. These results suggest that several loci in the rat brain should be active during the LT running. Especially, hypothalamic AVP neuron would be excited reflexively in the running. This would propose that running exercise at the moderate intensity could stimulate several region in the central nervous system especially in hypothalamus.Finally, we tried to examine how the MN innervating m.soleus (MN-SOL) and m.plantaris (MN-PL) could be recruited in the LT running by use of in situ hybridization for c-fos. Although the extent of c-fos expression in the MN-SOL was unchanged, that of MN-PL was increased just above the LT running, suggesting that these methods to detect c-fos gene/protein are available for exploring the active loci in the central nervous system in rat with running.