Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures

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Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures. / Gómez, Carlos Daniel; Rasmussen, Camilla Mai; Rekling, Jens C.

In: eNeuro, Vol. 8, No. 4, ENEURO.0154-21.2021, 2021.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Gómez, CD, Rasmussen, CM & Rekling, JC 2021, 'Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures', eNeuro, vol. 8, no. 4, ENEURO.0154-21.2021. https://doi.org/10.1523/ENEURO.0154-21.2021

APA

Gómez, C. D., Rasmussen, C. M., & Rekling, J. C. (2021). Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures. eNeuro, 8(4), [ENEURO.0154-21.2021]. https://doi.org/10.1523/ENEURO.0154-21.2021

Vancouver

Gómez CD, Rasmussen CM, Rekling JC. Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures. eNeuro. 2021;8(4). ENEURO.0154-21.2021. https://doi.org/10.1523/ENEURO.0154-21.2021

Author

Gómez, Carlos Daniel ; Rasmussen, Camilla Mai ; Rekling, Jens C. / Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures. In: eNeuro. 2021 ; Vol. 8, No. 4.

Bibtex

@article{e3b2aa1aff9f43a7a24c283c834fe929,
title = "Gabaergic inhibition of presynaptic ca2+ transients in respiratory preb{\"o}tzinger neurons in organotypic slice cultures",
abstract = "GABAergic somatodendritic inhibition in the preB{\"o}tzinger complex (preB{\"o}tC), a medullary site for the generation of inspiratory rhythm, is involved in respiratory rhythmogenesis and patterning. Nevertheless, whether GABA acts distally on presynaptic terminals, evoking presynaptic inhibition is unknown. Here, we begin to address this problem by measuring presynaptic Ca2+ transients in preB{\"o}tC neurons, under rhythmic and non-rhythmic conditions, with two variants of genetically encoded Ca2+ indicators (GECIs). Organotypic slice cultures from newborn mice, containing the preB{\"o}tC, were drop-transduced with jGCaMP7s, or injected with jGCaMP7f-labeling commissural preB{\"o}tC neurons. Then, Ca2+ imaging combined with whole-cell patch-clamp or field stimulation was obtained from inspiratory preB{\"o}tC neurons. We found that rhythmically active neurons expressed synchronized Ca2+ transients in soma, proximal and distal dendritic regions, and punctate synapse-like structures. Expansion microscopy revealed morphologic characteristics of bona fide synaptic boutons of the en passant and terminal type. Under non-rhythmic conditions, we found that bath application of the GABAA receptor agonist muscimol, and local microiontophoresis of GABA, reduced action potential (AP)-evoked and field stimulus-evoked Ca2+ transients in presynaptic terminals in inspiratory neurons and commissural neurons projecting to the contralateral preB{\"o}tC. In addition, under rhythmic conditions, network rhythmic activity was suppressed by muscimol, while the GABAA receptor antagonist bicuculline completely re-activated spontaneous activity. These observations demonstrate that the preB{\"o}tC includes neurons that show GABAergic inhibition of presynaptic Ca2+ transients, and presynaptic inhibition may play a role in the network activity that underlies breathing.",
keywords = "Breathing, GABA, GECI, JGCaMP7, Presynaptic inhibition",
author = "G{\'o}mez, {Carlos Daniel} and Rasmussen, {Camilla Mai} and Rekling, {Jens C.}",
note = "Publisher Copyright: {\textcopyright} 2021 Daniel G{\'o}mez et al.",
year = "2021",
doi = "10.1523/ENEURO.0154-21.2021",
language = "English",
volume = "8",
journal = "eNeuro",
issn = "2373-2822",
publisher = "Society for Neuroscience",
number = "4",

}

RIS

TY - JOUR

T1 - Gabaergic inhibition of presynaptic ca2+ transients in respiratory prebötzinger neurons in organotypic slice cultures

AU - Gómez, Carlos Daniel

AU - Rasmussen, Camilla Mai

AU - Rekling, Jens C.

N1 - Publisher Copyright: © 2021 Daniel Gómez et al.

PY - 2021

Y1 - 2021

N2 - GABAergic somatodendritic inhibition in the preBötzinger complex (preBötC), a medullary site for the generation of inspiratory rhythm, is involved in respiratory rhythmogenesis and patterning. Nevertheless, whether GABA acts distally on presynaptic terminals, evoking presynaptic inhibition is unknown. Here, we begin to address this problem by measuring presynaptic Ca2+ transients in preBötC neurons, under rhythmic and non-rhythmic conditions, with two variants of genetically encoded Ca2+ indicators (GECIs). Organotypic slice cultures from newborn mice, containing the preBötC, were drop-transduced with jGCaMP7s, or injected with jGCaMP7f-labeling commissural preBötC neurons. Then, Ca2+ imaging combined with whole-cell patch-clamp or field stimulation was obtained from inspiratory preBötC neurons. We found that rhythmically active neurons expressed synchronized Ca2+ transients in soma, proximal and distal dendritic regions, and punctate synapse-like structures. Expansion microscopy revealed morphologic characteristics of bona fide synaptic boutons of the en passant and terminal type. Under non-rhythmic conditions, we found that bath application of the GABAA receptor agonist muscimol, and local microiontophoresis of GABA, reduced action potential (AP)-evoked and field stimulus-evoked Ca2+ transients in presynaptic terminals in inspiratory neurons and commissural neurons projecting to the contralateral preBötC. In addition, under rhythmic conditions, network rhythmic activity was suppressed by muscimol, while the GABAA receptor antagonist bicuculline completely re-activated spontaneous activity. These observations demonstrate that the preBötC includes neurons that show GABAergic inhibition of presynaptic Ca2+ transients, and presynaptic inhibition may play a role in the network activity that underlies breathing.

AB - GABAergic somatodendritic inhibition in the preBötzinger complex (preBötC), a medullary site for the generation of inspiratory rhythm, is involved in respiratory rhythmogenesis and patterning. Nevertheless, whether GABA acts distally on presynaptic terminals, evoking presynaptic inhibition is unknown. Here, we begin to address this problem by measuring presynaptic Ca2+ transients in preBötC neurons, under rhythmic and non-rhythmic conditions, with two variants of genetically encoded Ca2+ indicators (GECIs). Organotypic slice cultures from newborn mice, containing the preBötC, were drop-transduced with jGCaMP7s, or injected with jGCaMP7f-labeling commissural preBötC neurons. Then, Ca2+ imaging combined with whole-cell patch-clamp or field stimulation was obtained from inspiratory preBötC neurons. We found that rhythmically active neurons expressed synchronized Ca2+ transients in soma, proximal and distal dendritic regions, and punctate synapse-like structures. Expansion microscopy revealed morphologic characteristics of bona fide synaptic boutons of the en passant and terminal type. Under non-rhythmic conditions, we found that bath application of the GABAA receptor agonist muscimol, and local microiontophoresis of GABA, reduced action potential (AP)-evoked and field stimulus-evoked Ca2+ transients in presynaptic terminals in inspiratory neurons and commissural neurons projecting to the contralateral preBötC. In addition, under rhythmic conditions, network rhythmic activity was suppressed by muscimol, while the GABAA receptor antagonist bicuculline completely re-activated spontaneous activity. These observations demonstrate that the preBötC includes neurons that show GABAergic inhibition of presynaptic Ca2+ transients, and presynaptic inhibition may play a role in the network activity that underlies breathing.

KW - Breathing

KW - GABA

KW - GECI

KW - JGCaMP7

KW - Presynaptic inhibition

U2 - 10.1523/ENEURO.0154-21.2021

DO - 10.1523/ENEURO.0154-21.2021

M3 - Journal article

C2 - 34380658

AN - SCOPUS:85113726305

VL - 8

JO - eNeuro

JF - eNeuro

SN - 2373-2822

IS - 4

M1 - ENEURO.0154-21.2021

ER -

ID: 279429004