Dept. of Neurobiology and Behavior, Seeley Mudd Hall, Cornell Univ., Ithaca NY 14853, USA.
J Neurophysiol. 2011 Nov;106(5):2783-9. doi: 10.1152/jn.00673.2011. Epub 2011 Sep 7.
It has been very difficult to record from interneurons in acute slices of the lumbar spinal cord from mice >3 wk of age. The low success rate and short recording times limit in vitro experimentation on mouse spinal networks to neonatal and early postnatal periods when locomotor networks are still developmentally immature. To overcome this limitation and enable investigation of mature locomotor network neurons, we have established a reliable procedure to record from spinal cord neurons in slices from adult, behaviorally mature mice of any age. Two key changes to the established neonate procedure were implemented. First, we remove the cord by a dorsal laminectomy from a deeply anesthetized animal. This enables respiration and other vital functions to continue up to the moment the maximally oxygenated lumbar spinal cord is removed, improving the health of the slices. Second, since adult spinal cord interneurons appear more sensitive to the intracellular dialysis that occurs during whole cell recordings, we introduced perforated patch recordings to the procedure. Stable recordings up to 12 h in duration were obtained with our new method. This will allow investigation of changes in mature neuronal properties in disease states or after spinal cord injury and allow prolonged recordings of responses to drug application that were previously impossible.
从年龄大于 3 周的小鼠的腰椎脊髓急性切片中记录中间神经元一直非常困难。低成功率和短记录时间限制了体外实验在运动网络仍处于发育不成熟的新生儿和早期新生儿期进行。为了克服这一限制并能够研究成熟的运动网络神经元，我们已经建立了一种可靠的程序，可以从任何年龄的行为成熟的成年小鼠的脊髓切片中记录脊髓神经元。对已建立的新生儿程序进行了两项关键更改。首先，我们通过对深度麻醉动物进行背侧椎板切除术切除脊髓。这可以使呼吸和其他重要功能持续到最大限度地供氧的腰椎脊髓被切除的那一刻，从而改善切片的健康状况。其次，由于成年脊髓中间神经元似乎对全细胞记录过程中发生的细胞内透析更为敏感，我们在该程序中引入了穿孔贴片记录。我们的新方法可获得长达 12 小时的稳定记录。这将允许研究疾病状态或脊髓损伤后成熟神经元特性的变化，并允许对以前不可能的药物应用反应进行长时间记录。
