Yamanaka Yuko, Kitamura Naoki, Shibuya Izumi
Department of Veterinary Physiology, Faculty of Agriculture, Tottori University, Koyama, Tottori, Japan.
Biomed Res. 2008 Aug;29(4):205-11. doi: 10.2220/biomedres.29.205.
Ten pairs of protrusions, called accessory lobes (ALs), exist at the lateral sides of avian lumbosacral spinal cords. Histological evidence has shown that neurons are present in AL and behavioral evidence suggests that AL acts as a sensory organ of equilibrium during bipedal walking. However, there is little functional evidence to indicate that cells in AL have neuronal functions. To elucidate this point, we developed a method to dissociate cells from chick AL and made electrophysiological recordings with the whole-cell patch clamp technique. Cells dissociated by enzymatic digestion from chick AL contained two major types of cells. One was round with clear cytosol and the other had a round cell body, rich cytosolic structures and some processes. Rapidly activating inward currents and slowly activating outward currents were recorded in response to depolarizing pulses to -10 mV under the voltage clamp configuration only from the latter type of cells. TTX at 100 nM inhibited the inward current by 85%, indicating the functional expression of TTX-sensitive voltage-gated Na(+) channel (VGSC). Activation and inactivation kinetics of the inward currents in AL cells were similar to those of mammalian VGSC. The VGSC-expressing AL cells generated action potentials in response to depolarization under the current clamp configuration. These results clearly indicate that functional neurons expressing fast inactivating and TTXsensitive VGSC which generate action potentials exist in the AL of the chick. These lines of cellular evidence clearly indicate that functional neurons exist in ALs and further support the proposal that the chick ALs function as the sensory organ of equilibrium.
鸟类腰骶脊髓的外侧存在十对突出物,称为副叶(ALs)。组织学证据表明副叶中存在神经元,行为学证据表明在双足行走过程中副叶充当平衡感觉器官。然而,几乎没有功能证据表明副叶中的细胞具有神经元功能。为阐明这一点,我们开发了一种从鸡副叶分离细胞的方法,并使用全细胞膜片钳技术进行电生理记录。通过酶消化从鸡副叶分离的细胞包含两种主要类型的细胞。一种是圆形的,胞质清晰,另一种有圆形的细胞体、丰富的胞质结构和一些突起。仅从后一种类型的细胞在电压钳模式下对 -10 mV 的去极化脉冲记录到快速激活的内向电流和缓慢激活的外向电流。100 nM 的河豚毒素(TTX)使内向电流抑制了 85%,表明 TTX 敏感的电压门控钠通道(VGSC)的功能表达。副叶细胞内向电流的激活和失活动力学与哺乳动物 VGSC 相似。在电流钳模式下,表达 VGSC 的副叶细胞对去极化产生动作电位。这些结果清楚地表明,在鸡的副叶中存在表达快速失活且对 TTX 敏感的 VGSC 并能产生动作电位的功能性神经元。这些细胞证据清楚地表明副叶中存在功能性神经元,并进一步支持了鸡副叶作为平衡感觉器官发挥功能的观点。