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Optogenetic Approach to Local Neuron Network Analysis of the Medullary Respiratory Center

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Optogenetics

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1293))

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Abstract

Using an optogenetic approach, we analyzed a local neuron network of the respiratory center in the medulla of a brainstem–spinal cord preparation isolated from neonatal rat. We developed a transgenic (Tg) rat line in which Phox2b-positive cells expressed archaerhodopsin-3 (Arch) or one of the step-function channelrhodopsin variants (ChRFR) under the control of Phox2b promoter-enhancer regions. Then, in en bloc preparations from 0- to 2-day-old Tg neonatal rats, we analyzed membrane potential changes of medullary respiratory-related neurons in response to photostimulation of the rostral ventral medulla. The photostimulation-induced inhibition or facilitation of the respiratory rhythm in Arch-expressing or ChRFR-expressing Tg rat preparations, respectively. Selective photoactivation of Phox2b-positive neurons expressing ChRFR in the rostral ventrolateral medulla of a neonatal rat en bloc preparation induced membrane potential changes of respiratory-related neurons that were dependent on heterogeneous properties of synaptic connections in the respiratory center. We concluded that the optogenetic approach is a powerful method of verifying a hypothetical model of local networks among respiratory-related neurons in the rostral ventrolateral medulla of neonatal rat.

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Abbreviations

AICA:

Anterior inferior cerebellar artery

Arch:

Archaerhodopsin-3

C4:

Fourth cervical ventral root

ChRFR:

Step-function channelrhodopsin variants

Exp:

Expiratory

EYFP:

Enhanced yellow fluorescent protein

FN:

Facial nucleus

Insp:

Inspiratory

LED:

Light-emitting diode

pFRG:

Parafacial respiratory group

preBötC:

preBötzinger complex

Pre-I:

Preinspiratory

RTN:

Retrotrapezoid nucleus

Tg:

Transgenic

TH:

Tyrosine hydroxylase

TTX:

Tetrodotoxin

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Acknowledgments

This research was supported by Scientific Research on Innovative Areas (Comprehensive Brain Science Network) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, and JSPS KAKENHI (16K07003, 25430012), and by the Program for the Strategic Research Foundation at Private Universities 2016–2017 (Showa University School of Medicine and Jichi Medical University).

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Authors and Affiliations

  1. Department of Physiology, Showa University School of Medicine, Tokyo, Japan

    Hiroshi Onimaru

  2. Clinical Research Department, National Hospital Organization Murayama Medical Center, Tokyo, Japan

    Keiko Ikeda

Authors
  1. Hiroshi Onimaru
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  2. Keiko Ikeda
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Correspondence to Hiroshi Onimaru .

Editor information

Editors and Affiliations

  1. The Institute for Solid State Physics, The University of Tokyo, Kashiwa, Japan

    Hiromu Yawo

  2. OptoBioTechnology Research Center, Nagoya Institute of Technology, Nagoya, Japan

    Hideki Kandori

  3. National Institutes of Natural Sciences, Tokyo, Japan

    Amane Koizumi

  4. Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan

    Ryoichiro Kageyama

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Onimaru, H., Ikeda, K. (2021). Optogenetic Approach to Local Neuron Network Analysis of the Medullary Respiratory Center. In: Yawo, H., Kandori, H., Koizumi, A., Kageyama, R. (eds) Optogenetics. Advances in Experimental Medicine and Biology, vol 1293. Springer, Singapore. https://doi.org/10.1007/978-981-15-8763-4_29

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