Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA 15213, United States.
Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA 15213, United States.
Neuroscience. 2020 Jan 15;425:12-28. doi: 10.1016/j.neuroscience.2019.11.013. Epub 2019 Nov 27.
Coordination of activity of external urethral sphincter (EUS) striated muscle and bladder (BL) smooth muscle is essential for efficient voiding. In this study we examined the morphological and electrophysiological properties of neurons in the L3/L4 spinal cord (SC) that are likely to have an important role in EUS-BL coordination in rats. EUS-related SC neurons were identified by retrograde transsynaptic tracing following injection of pseudorabies virus (PRV) co-expressing fluorescent markers into the EUS of P18-P20 male rats. Tracing revealed not only EUS motoneurons in L6/S1 but also interneurons in lamina X of the L6/S1 and L3/L4 SC. Physiological properties of fluorescently labeled neurons were assessed during whole-cell recordings in SC slices followed by reconstruction of biocytin-filled neurons. Reconstructions of neuronal processes from transverse or longitudinal slices showed that some L3/L4 neurons have axons projecting toward and into the ventro-medial funiculus (VMf) where axons extended caudally. Other neurons had axons projecting within laminae X and VII. Dendrites of L3/L4 neurons were distributed within laminae X and VII. The majority of L3/L4 neurons exhibited tonic firing in response to depolarizing currents. In transverse slices focal electrical stimulation (FES) in the VMf or in laminae X and VII elicited antidromic axonal spikes and/or excitatory synaptic responses in L3/L4 neurons; while in longitudinal slices FES elicited excitatory synaptic inputs from sites up to 400 μm along the central canal. Inhibitory inputs were rarely observed. These data suggest that L3/L4 EUS-related circuitry consists of at least two neuronal populations: segmental interneurons and propriospinal neurons projecting to L6/S1.
外尿道括约肌(EUS)横纹肌和膀胱(BL)平滑肌的活动协调对于有效排尿至关重要。在这项研究中,我们检查了 L3/L4 脊髓(SC)中神经元的形态和电生理特性,这些神经元可能在大鼠 EUS-BL 协调中发挥重要作用。通过将表达荧光标记物的伪狂犬病毒(PRV)注入 P18-P20 雄性大鼠的 EUS 后,逆行顺行追踪鉴定与 EUS 相关的 SC 神经元。追踪不仅显示了 L6/S1 中的 EUS 运动神经元,还显示了 L6/S1 和 L3/L4 SC 中 X 层的中间神经元。在 SC 切片中的全细胞记录期间评估了荧光标记神经元的生理特性,然后对生物胞素填充神经元进行重建。从横切片或纵切片重建神经元过程表明,一些 L3/L4 神经元的轴突向并进入腹内侧束(VMf)投射,并向尾部延伸。其他神经元的轴突在 X 和 VII 层内投射。L3/L4 神经元的树突分布在 X 和 VII 层内。大多数 L3/L4 神经元在响应去极化电流时表现出紧张性放电。在横切片中,VMf 或 X 和 VII 层中的聚焦电刺激(FES)在 L3/L4 神经元中诱发轴突逆行尖峰和/或兴奋性突触反应;而在纵切片中,FES 诱发来自中央管沿长轴达 400 μm 处的兴奋性突触输入。很少观察到抑制性输入。这些数据表明,L3/L4 EUS 相关回路至少由两个神经元群体组成:节段性中间神经元和投射到 L6/S1 的 propriospinal 神经元。