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

我们的目的是描绘以乳酸阈值（LT）速度跑步的大鼠大脑中的活动部位。因此，我们使用我们的跑步模型，其中雄性大鼠在跑步机上。在LT运行期间和之后，取出全脑并将其额叶切片用于c-fos的原位杂交（实验1）。此外，我们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，脑中c-fos的表达。实验中采用免疫组化方法检测c-fos蛋白的表达。本实验还观察了c-fos阳性细胞内精氨酸加压素（AVP）的免疫反应性。在实验Ⅰ中，c-fos mRNA在下丘脑（室旁核和视上核）、丘脑、海马、边缘区等部位以及顶叶等运动中枢表达增加。实验II中，AVP细胞中出现c-fos阳性染色，提示AVP神经元参与了运动应激。c-fos蛋白仅在运动压力反应时才在AVP阳性神经元中表达。这些结果表明，在LT运行期间，大鼠大脑中的几个位点应该是活跃的。特别是下丘脑AVP神经元在运动中会反射性兴奋。最后，我们尝试用c-fos原位杂交技术研究了运动训练对比目鱼肌和跖肌神经元（MN-SOL和MN-PL）的影响。MN-SOL的c-fos表达水平无明显变化，而MN-PL的c-fos表达水平略高于LT，提示本研究建立的c-fos基因/蛋白检测方法可用于探索运动大鼠中枢神经系统的活动部位。

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.