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Difference of acute dissociation and 1-day culture on the electrophysiological properties of rat dorsal root ganglion neurons

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Abstract

The dissociated dorsal root ganglion (DRG) neurons with or without culture were widely used for investigation of their electrophysiological properties. The culture procedures, however, may alter the properties of these neurons and the effects are not clear. In the present study, we recorded the action potentials (AP) and the voltage-gated Na+, K+, and Ca2+ currents with patch clamp technique and measured the mRNA of Nav1.6–1.9 and Cav2.1–2.2 with real-time PCR technique from acutely dissociated and 1-day (1-d) cultured DRG neurons. The effects of the nerve growth factor (NGF) on the expression of Nav1.6–1.9 and Cav2.1–2.2 were evaluated. The neurons were classified as small (DRG-S), medium (DRG-M), and large (DRG-L), according to their size frequency distribution pattern. We found 1-d culture increased the AP size but reduced the excitability, and reduced the voltage-gated Na+ and Ca2+ currents and their corresponding mRNA expression in all types of neurons. The lack of NGF in the culture medium may contribute to the reduced Na+ and Ca2+ current, as the application of NGF recovered some of the reduced transcripts (Nav1.9, Cav2.1, and Cav2.2). 1-d culture showed neuron-type specific effects on some of the AP properties: it increased the maximum AP depolarizing rate (MDR) and hyperpolarized the resting membrane potential (RP) in DRG-M and DRG-L neurons, but slowed the maximum AP repolarizing rate (MRR) in DRG-S neurons. In conclusion, the 1-d cultured neurons had different properties with those of the acutely dissociated neurons, and lack of NGF may contribute to some of these differences.

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Abbreviations

AP:

Action potential

APD50:

AP duration at 50% AP size

DRG-S:

Small-sized DRG neurons

DRG-M:

Medium-sized DRG neurons

DRG-L:

Large-sized DRG neurons

MDR:

Maximum depolarizing rate of AP

MRR:

Maximum repolarizing rate of AP

I Na-s :

INa with slow component only

I Na-f :

INa with fast component only

I Na-m :

INa with mixed fast and slow components

I Ki :

Inward K+ current

I Ko :

Outward K+ current

R in :

Input resistance

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Acknowledgements

We thank Prof. Sally N Lawson in the University of Bristol for the advice of experiment design.

Funding

This work is supported by grants from National Nature Science Foundation of China (NSFC Grant No.:81100818 and No.:30670854) to Linlin Gao.

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Authors and Affiliations

  1. Department of Physiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Rd., Wuhan, China

    Yuanlong Song, Miaomiao Zhang, Xiaoqing Tao, Zifen Xu, Yunjie Zheng, Minjie Zhu, Liangpin Zhang, Jinhan Qiao & Linlin Gao

  2. Institutes of Brain Research, Huazhong University of Science and Technology, Wuhan, China

    Yuanlong Song & Linlin Gao

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Song, Y., Zhang, M., Tao, X. et al. Difference of acute dissociation and 1-day culture on the electrophysiological properties of rat dorsal root ganglion neurons. J Physiol Biochem 74, 207–221 (2018). https://doi.org/10.1007/s13105-017-0606-9

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