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

我们的目的是描绘大鼠大脑在速度@乳酸阈值（LT）下的活动部位。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，切除全脑并在额叶切片进行c-fos原位杂交（exp1）。此外，我们进一步研究了在*20运动阈值条件下，坐骨神经电刺激引起肌肉收缩后大脑如何表达c-fos。本实验采用免疫组化方法检测c-fos蛋白。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。在大鼠中，c-fos mRNA在下丘脑（室旁核和视上核）、丘脑、海马、边缘区以及运动中心和顶叶区均有表达。在Exp.II中，AVP细胞c-fos染色呈阳性，提示AVP神经元参与了运动应激。表达AVP的神经元细胞中c-fos蛋白的表达仅在发生“运动压力反应”的情况下观察到。这些结果表明，大鼠大脑中的几个位点在LT跑步期间应该是活跃的。特别是下丘脑AVP神经元在跑步过程中会反射性兴奋。这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，特别是下丘脑。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plantaris （MN- pl）的MN是如何在LT运动中被招募的。虽然MN-SOL中c-fos的表达量没有变化，但MN-PL中c-fos的表达量仅高于LT跑步时，这表明这些检测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位点。

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.