Li Bai-Yan, Schild John H
Department of Biomedical Engineering, Indiana University Purdue University Indianapolis, Indianapolis, IN 46202, USA.
J Neurosci Methods. 2007 Aug 15;164(1):75-85. doi: 10.1016/j.jneumeth.2007.04.003. Epub 2007 Apr 8.
An unavoidable consequence of enzymatic dispersion of sensory neurons from intact ganglia is loss of the axon and thus the ability to classify afferent fiber type based upon conduction velocity (CV). An intact rat nodose ganglion preparation was used to randomly sample neurons (n=76) using the patch clamp technique. Reliable electrophysiological and chemophysiological correlates of afferent fiber type were established for use with isolated neuron preparations. Myelinated afferents (approximately 25%) formed two groups exhibiting strikingly different functional profiles. One group (n=10) exhibited CVs in excess of 10 m/s and narrow (<1 ms) action potentials (APs) while the other (n=9) had CVs as low as 4m/s and broad (>2 ms) APs that closely approximated those identified as unmyelinated afferents (n=57) with CVs less than 1m/s. A cluster analysis of select measures from the AP waveforms strongly correlated with CV, producing three statistically unique populations (p<0.05). These groupings aligned with our earlier hypothesis (Jin et al., 2004) that a differential sensitivity to the selective purinergic and vanilloid receptor agonists can be used as reliable pharmacological indicators of vagal afferent fiber type. These metrics were further validated using an even larger population of isolated (n=240) nodose neurons. Collectively, these indicators of afferent fiber type can be used to provide valuable insight concerning the relavence of isolated cellular observations to integrated afferent function of visceral organ systems.
从完整神经节中酶解分散感觉神经元不可避免的后果是轴突丧失,从而失去了根据传导速度(CV)对传入纤维类型进行分类的能力。使用完整的大鼠结状神经节标本,采用膜片钳技术随机采样神经元(n = 76)。建立了与传入纤维类型相关的可靠电生理和化学生理关联,用于分离的神经元标本。有髓传入纤维(约25%)形成两组,表现出截然不同的功能特征。一组(n = 10)的传导速度超过10 m/s,动作电位(AP)窄(<1 ms),而另一组(n = 9)的传导速度低至4m/s,动作电位宽(>2 ms),与鉴定为无髓传入纤维(n = 57)的动作电位非常相似,其传导速度小于1m/s。对动作电位波形的选定测量值进行聚类分析,与传导速度密切相关,产生了三个统计学上独特的群体(p<0.05)。这些分组与我们早期的假设(Jin等人,2004年)一致,即对选择性嘌呤能和香草酸受体激动剂的不同敏感性可作为迷走神经传入纤维类型的可靠药理学指标。使用更大数量的分离(n = 240)结状神经元进一步验证了这些指标。总体而言,这些传入纤维类型指标可用于提供有价值的见解,说明分离的细胞观察结果与内脏器官系统整合传入功能的相关性。