Song Z-M, Hu J, Rudy B, Redman S J
Medical School, Australian National University, Canberra, ACT, Australia.
Neuroscience. 2006 May 12;139(2):531-8. doi: 10.1016/j.neuroscience.2005.12.048. Epub 2006 Feb 7.
One class of spinal interneurons, the Renshaw cells, is able to discharge at very high frequencies in adult mammals. Neuronal firing at such high frequencies requires voltage-gated potassium channels to rapidly repolarize the membrane potential after each action potential. We sought to establish the pattern of expression of calbindin and potassium channels with Kv3.1b and Kv3.2 subunits in Renshaw cells at different developmental stages of postnatal mice. The pattern of expression of calbindin changed dramatically during early postnatal development. An adult pattern of calbindin reactive neurons started to emerge from postnatal day 10 to postnatal day 14, with cells in laminae I and II of superficial dorsal horn and the ventral lamina VII. Renshaw cells were identified immunohistochemically by their expression of calbindin and their location in the ventral horn of the spinal cord. Western blot results of the lumbar spinal cord showed that Kv3.1b expression became faintly evident from postnatal day 10, reached a maximum at postnatal day 21 and was maintained through postnatal day 49. Double labeling results showed that all Renshaw cells expressed Kv3.1b weakly from postnatal day 14, and strongly at postnatal day 21. Western blot results showed that Kv3.2 expression became detectable in the lumbar cord from postnatal day 12, and increased steadily until reaching an adult level at postnatal day 28. In contrast to the Kv3.1b results, Kv3.2 was not expressed in Renshaw cells, although some neurons located at laminae VIII and VI expressed Kv3.2. We conclude that Renshaw cells express Kv3.1b but not Kv3.2 from postnatal day 14.
一类脊髓中间神经元,即闰绍细胞,在成年哺乳动物中能够以非常高的频率放电。神经元以如此高的频率放电需要电压门控钾通道在每次动作电位后迅速使膜电位复极化。我们试图确定出生后小鼠不同发育阶段闰绍细胞中钙结合蛋白以及具有Kv3.1b和Kv3.2亚基的钾通道的表达模式。钙结合蛋白的表达模式在出生后早期发育过程中发生了显著变化。从出生后第10天到第14天,钙结合蛋白反应性神经元的成年模式开始出现,位于浅表背角的I层和II层以及腹侧VII层。通过闰绍细胞中钙结合蛋白的表达及其在脊髓腹角的位置进行免疫组织化学鉴定。腰段脊髓的蛋白质免疫印迹结果显示,Kv3.1b的表达从出生后第10天开始隐约可见,在出生后第21天达到最大值,并持续到出生后第49天。双重标记结果显示,所有闰绍细胞从出生后第14天开始弱表达Kv3.1b,在出生后第21天强烈表达。蛋白质免疫印迹结果显示,Kv3.2的表达从出生后第12天开始在腰段脊髓中可检测到,并持续稳定增加,直到出生后第28天达到成年水平。与Kv3.1b的结果相反,闰绍细胞中不表达Kv3.2,尽管位于VIII层和VI层的一些神经元表达Kv3.2。我们得出结论,闰绍细胞从出生后第14天开始表达Kv3.1b而不表达Kv3.2。