Axonal CB1 Receptors Mediate Inhibitory Bouton Formation via cAMP Increase and PKA

Jian Liang; Dennis L.H. Kruijssen; Aniek C.J. Verschuuren; Bas J.B. Voesenek; Feline F.W. Benavides; Maria Sáez Gonzalez; Marvin Ruiter; Corette J. Wierenga

doi:10.1523/jneurosci.0851-21.2021

Axonal CB1 Receptors Mediate Inhibitory Bouton Formation via cAMP Increase and PKA

Jian Liang, Dennis L.H. Kruijssen, Aniek C.J. Verschuuren, Bas J.B. Voesenek, Feline F.W. Benavides, Maria Sáez Gonzalez, Marvin Ruiter, Corette J. Wierenga^*

^*Corresponding author for this work

Utrecht University

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Experience-dependent formation and removal of inhibitory synapses are essential throughout life. For instance, GABAergic synapses are removed to facilitate learning, and strong excitatory activity is accompanied by the formation of inhibitory synapses to maintain coordination between excitation and inhibition. We recently discovered that active dendrites trigger the growth of inhibitory synapses via CB1 receptor-mediated endocannabinoid signaling, but the underlying mechanism remained unclear. Using two-photon microscopy to monitor the formation of individual inhibitory boutons in hippocampal organotypic slices from mice (both sexes), we found that CB1 receptor activation mediated the formation of inhibitory boutons and promoted their subsequent stabilization. Inhibitory bouton formation did not require neuronal activity and was independent of Gi/o-protein signaling, but was directly induced by elevating cAMP levels using forskolin and by activating Gs-proteins using DREADDs. Blocking PKA activity prevented CB1 receptor-mediated inhibitory bouton formation. Our findings reveal that axonal CB1 receptors signal via unconventional downstream pathways and that inhibitory bouton formation is triggered by an increase in axonal cAMP levels. Our results demonstrate an unexpected role for axonal CB1 receptors in axon-specific, and context-dependent, inhibitory synapse formation.

Original language	English
Pages (from-to)	8279-8296
Number of pages	18
Journal	Journal of Neuroscience
Volume	41
Issue number	40
DOIs	https://doi.org/10.1523/jneurosci.0851-21.2021
Publication status	Published - 6 Oct 2021

Bibliographical note

Funding Information:
This work was supported by a CSC scholarship (J.L.), by Alzheimer Nederland (Grant WE.03-2018-11, to M.R.) and by the Netherlands Organization for Scientific Research (Grant OCENW.KLEIN.150, to M.S.G.), and as part of the research program of the Foundation for Fundamental Research on Matter (Grants #15PR3178 and #16NEPH05, to D.L.H.K.). We thank Lotte Herstel for helping with experiments shown in Figure 1, E and F, and René van Dorland for excellent technical support. The authors declare no competing financial interests. Correspondence should be addressed to Corette J. Wierenga at [email protected]. https://doi.org/10.1523/JNEUROSCI.0851-21.2021 Copyright © 2021 the authors

Publisher Copyright:
© 2021 the authors.

Funding

This work was supported by a CSC scholarship (J.L.), by Alzheimer Nederland (Grant WE.03-2018-11, to M.R.) and by the Netherlands Organization for Scientific Research (Grant OCENW.KLEIN.150, to M.S.G.), and as part of the research program of the Foundation for Fundamental Research on Matter (Grants #15PR3178 and #16NEPH05, to D.L.H.K.). We thank Lotte Herstel for helping with experiments shown in Figure 1, E and F, and René van Dorland for excellent technical support. The authors declare no competing financial interests. Correspondence should be addressed to Corette J. Wierenga at [email protected]. https://doi.org/10.1523/JNEUROSCI.0851-21.2021 Copyright © 2021 the authors

Keywords

cAMP/PKA
endocannabinoids
G-protein-coupled receptors
inhibitory synapses
synapse formation

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10.1523/jneurosci.0851-21.2021

8279.fullFinal published version, 2.7 MBLicence: Taverne

Cite this

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title = "Axonal CB1 Receptors Mediate Inhibitory Bouton Formation via cAMP Increase and PKA",

abstract = "Experience-dependent formation and removal of inhibitory synapses are essential throughout life. For instance, GABAergic synapses are removed to facilitate learning, and strong excitatory activity is accompanied by the formation of inhibitory synapses to maintain coordination between excitation and inhibition. We recently discovered that active dendrites trigger the growth of inhibitory synapses via CB1 receptor-mediated endocannabinoid signaling, but the underlying mechanism remained unclear. Using two-photon microscopy to monitor the formation of individual inhibitory boutons in hippocampal organotypic slices from mice (both sexes), we found that CB1 receptor activation mediated the formation of inhibitory boutons and promoted their subsequent stabilization. Inhibitory bouton formation did not require neuronal activity and was independent of Gi/o-protein signaling, but was directly induced by elevating cAMP levels using forskolin and by activating Gs-proteins using DREADDs. Blocking PKA activity prevented CB1 receptor-mediated inhibitory bouton formation. Our findings reveal that axonal CB1 receptors signal via unconventional downstream pathways and that inhibitory bouton formation is triggered by an increase in axonal cAMP levels. Our results demonstrate an unexpected role for axonal CB1 receptors in axon-specific, and context-dependent, inhibitory synapse formation. ",

keywords = "cAMP/PKA, endocannabinoids, G-protein-coupled receptors, inhibitory synapses, synapse formation",

author = "Jian Liang and Kruijssen, \{Dennis L.H.\} and Verschuuren, \{Aniek C.J.\} and Voesenek, \{Bas J.B.\} and Benavides, \{Feline F.W.\} and Gonzalez, \{Maria S{\'a}ez\} and Marvin Ruiter and Wierenga, \{Corette J.\}",

note = "Funding Information: This work was supported by a CSC scholarship (J.L.), by Alzheimer Nederland (Grant WE.03-2018-11, to M.R.) and by the Netherlands Organization for Scientific Research (Grant OCENW.KLEIN.150, to M.S.G.), and as part of the research program of the Foundation for Fundamental Research on Matter (Grants \#15PR3178 and \#16NEPH05, to D.L.H.K.). We thank Lotte Herstel for helping with experiments shown in Figure 1, E and F, and Ren{\'e} van Dorland for excellent technical support. The authors declare no competing financial interests. Correspondence should be addressed to Corette J. Wierenga at [email protected]. https://doi.org/10.1523/JNEUROSCI.0851-21.2021 Copyright {\textcopyright} 2021 the authors Publisher Copyright: {\textcopyright} 2021 the authors.",

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doi = "10.1523/jneurosci.0851-21.2021",

language = "English",

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AU - Liang, Jian

AU - Kruijssen, Dennis L.H.

AU - Verschuuren, Aniek C.J.

AU - Voesenek, Bas J.B.

AU - Benavides, Feline F.W.

AU - Gonzalez, Maria Sáez

AU - Ruiter, Marvin

AU - Wierenga, Corette J.

N1 - Funding Information: This work was supported by a CSC scholarship (J.L.), by Alzheimer Nederland (Grant WE.03-2018-11, to M.R.) and by the Netherlands Organization for Scientific Research (Grant OCENW.KLEIN.150, to M.S.G.), and as part of the research program of the Foundation for Fundamental Research on Matter (Grants #15PR3178 and #16NEPH05, to D.L.H.K.). We thank Lotte Herstel for helping with experiments shown in Figure 1, E and F, and René van Dorland for excellent technical support. The authors declare no competing financial interests. Correspondence should be addressed to Corette J. Wierenga at [email protected]. https://doi.org/10.1523/JNEUROSCI.0851-21.2021 Copyright © 2021 the authors Publisher Copyright: © 2021 the authors.

PY - 2021/10/6

Y1 - 2021/10/6

N2 - Experience-dependent formation and removal of inhibitory synapses are essential throughout life. For instance, GABAergic synapses are removed to facilitate learning, and strong excitatory activity is accompanied by the formation of inhibitory synapses to maintain coordination between excitation and inhibition. We recently discovered that active dendrites trigger the growth of inhibitory synapses via CB1 receptor-mediated endocannabinoid signaling, but the underlying mechanism remained unclear. Using two-photon microscopy to monitor the formation of individual inhibitory boutons in hippocampal organotypic slices from mice (both sexes), we found that CB1 receptor activation mediated the formation of inhibitory boutons and promoted their subsequent stabilization. Inhibitory bouton formation did not require neuronal activity and was independent of Gi/o-protein signaling, but was directly induced by elevating cAMP levels using forskolin and by activating Gs-proteins using DREADDs. Blocking PKA activity prevented CB1 receptor-mediated inhibitory bouton formation. Our findings reveal that axonal CB1 receptors signal via unconventional downstream pathways and that inhibitory bouton formation is triggered by an increase in axonal cAMP levels. Our results demonstrate an unexpected role for axonal CB1 receptors in axon-specific, and context-dependent, inhibitory synapse formation.

AB - Experience-dependent formation and removal of inhibitory synapses are essential throughout life. For instance, GABAergic synapses are removed to facilitate learning, and strong excitatory activity is accompanied by the formation of inhibitory synapses to maintain coordination between excitation and inhibition. We recently discovered that active dendrites trigger the growth of inhibitory synapses via CB1 receptor-mediated endocannabinoid signaling, but the underlying mechanism remained unclear. Using two-photon microscopy to monitor the formation of individual inhibitory boutons in hippocampal organotypic slices from mice (both sexes), we found that CB1 receptor activation mediated the formation of inhibitory boutons and promoted their subsequent stabilization. Inhibitory bouton formation did not require neuronal activity and was independent of Gi/o-protein signaling, but was directly induced by elevating cAMP levels using forskolin and by activating Gs-proteins using DREADDs. Blocking PKA activity prevented CB1 receptor-mediated inhibitory bouton formation. Our findings reveal that axonal CB1 receptors signal via unconventional downstream pathways and that inhibitory bouton formation is triggered by an increase in axonal cAMP levels. Our results demonstrate an unexpected role for axonal CB1 receptors in axon-specific, and context-dependent, inhibitory synapse formation.

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KW - endocannabinoids

KW - G-protein-coupled receptors

KW - inhibitory synapses

KW - synapse formation

UR - https://www.scopus.com/pages/publications/85117093877

U2 - 10.1523/jneurosci.0851-21.2021

DO - 10.1523/jneurosci.0851-21.2021

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SN - 0270-6474

VL - 41

SP - 8279

EP - 8296

JO - Journal of Neuroscience

JF - Journal of Neuroscience

IS - 40

ER -

Axonal CB1 Receptors Mediate Inhibitory Bouton Formation via cAMP Increase and PKA

Abstract

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Keywords

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