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Int J Biol Sci 2013; 9(7):716-727. doi:10.7150/ijbs.7006

Research Paper

Subtype Identification in Acutely Dissociated Rat Nodose Ganglion Neurons Based on Morphologic Parameters

Xiao-Long Lu*1,4, Wen-Xiao Xu*2, Zhen-Yu Yan1, Zhao Qian1, Bing Xu1, Yang Liu3, Li-Min Han1, Rui-Chen Gao3, Jun-Nan Li1, Mei Yuan1, Chong-Bao Zhao3, Guo-fen Qiao1,3✉, Bai-Yan Li1✉

1. Department of Pharmacology, Harbin Medical University, Harbin, China
2. Department of Orthopedics, the First Affiliated Hospital, Harbin Medical University, Harbin, China
3. Key Laboratory of Cardiovascular Medicine Research, Ministry of Education, Harbin Medical University, Harbin, China
4. Riley Heart Research Center, Division of Pediatric Cardiology, Herman B. Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, Indianapolis, USA
* These authors contributed equally to this work.

Abstract

Nodose ganglia are composed of A-, Ah- and C-type neurons. Despite their important roles in regulating visceral afferent function, including cardiovascular, pulmonary, and gastrointestinal homeostasis, information about subtype-specific expression, molecular identity, and function of individual ion transporting proteins is scarce. Although experiments utilizing the sliced ganglion preparation have provided valuable insights into the electrophysiological properties of nodose ganglion neuron subtypes, detailed characterization of their electrical phenotypes will require measurements in isolated cells. One major unresolved problem, however, is the difficulty to unambiguously identify the subtype of isolated nodose ganglion neurons without current-clamp recording, because the magnitude of conduction velocity in the corresponding afferent fiber, a reliable marker to discriminate subtypes in situ, can no longer be determined. Here, we present data supporting the notion that application of an algorithm regarding to microscopic structural characteristics, such as neuron shape evaluated by the ratio between shortest and longest axis, neuron surface characteristics, like membrane roughness, and axon attachment, enables specific and sensitive subtype identification of acutely dissociated rat nodose ganglion neurons, by which the accuracy of identification is further validated by electrophysiological markers and overall positive predictive rates is 89.26% (90.04%, 76.47%, and 98.21% for A-, Ah, and C-type, respectively). This approach should aid in gaining insight into the molecular correlates underlying phenotypic heterogeneity of nodose ganglia. Additionally, several critical points that help for neuron identification and afferent conduction calibration are also discussed.

Keywords: visualization, visceral sensory neuron, conduction velocity, whole-cell patch, action potential

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Lu XL, Xu WX, Yan ZY, Qian Z, Xu B, Liu Y, Han LM, Gao RC, Li JN, Yuan M, Zhao CB, Qiao Gf, Li BY. Subtype Identification in Acutely Dissociated Rat Nodose Ganglion Neurons Based on Morphologic Parameters. Int J Biol Sci 2013; 9(7):716-727. doi:10.7150/ijbs.7006. Available from http://www.ijbs.com/v09p0716.htm