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(A) Schematics of a DRN slice and positioning of recording <t>electrode</t> and optic fiber in Vip-Cre/COP4 mice. (B) Confocal images of a patched GFP+ neuron filled with biocytin in the DRN of a Vip-Cre/COP4 mouse (C) Channelrhodopsin evoked currents in a DRN GFP+ neuron associated to fluorescent and infra-red images (D) Schematics of ovBNST and CeA slices and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (E) Confocal images of patched neurons filled with biocytin and PKC-δ immunostaining in ovBNST and CeA. (F) Excitatory post-synaptic evoked currents recorded in ovBNST and CeA neurons upon optogenetic stimulation of DRN VIP fibers. Purple curves show inward currents abolished by application NMDA antagonists, APV and CNQX (black curve). (G) Donut graphs showing the proportion of neurons responding to optogenetic stimulation of DRN VIP fibers colocalizing with PKC-δ. (H) Ex vivo electrophysiological characterization of ovBNST and CeA excitatory post-synaptic currents evoked by optogenetic stimulation of DRN VIP fibers. Abbreviations: DRN: dorsal raphe nucleus; ovBNST oval nucleus of the bed nucleus of the stria terminalis; Aq: aqueduct of Sylvius; CeA: central nucleus of the amygdala; PPR: paired-pulse ratio.
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(A) Schematics of a DRN slice and positioning of recording <t>electrode</t> and optic fiber in Vip-Cre/COP4 mice. (B) Confocal images of a patched GFP+ neuron filled with biocytin in the DRN of a Vip-Cre/COP4 mouse (C) Channelrhodopsin evoked currents in a DRN GFP+ neuron associated to fluorescent and infra-red images (D) Schematics of ovBNST and CeA slices and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (E) Confocal images of patched neurons filled with biocytin and PKC-δ immunostaining in ovBNST and CeA. (F) Excitatory post-synaptic evoked currents recorded in ovBNST and CeA neurons upon optogenetic stimulation of DRN VIP fibers. Purple curves show inward currents abolished by application NMDA antagonists, APV and CNQX (black curve). (G) Donut graphs showing the proportion of neurons responding to optogenetic stimulation of DRN VIP fibers colocalizing with PKC-δ. (H) Ex vivo electrophysiological characterization of ovBNST and CeA excitatory post-synaptic currents evoked by optogenetic stimulation of DRN VIP fibers. Abbreviations: DRN: dorsal raphe nucleus; ovBNST oval nucleus of the bed nucleus of the stria terminalis; Aq: aqueduct of Sylvius; CeA: central nucleus of the amygdala; PPR: paired-pulse ratio.
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(A) Schematics of a DRN slice and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (B) Confocal images of a patched GFP+ neuron filled with biocytin in the DRN of a Vip-Cre/COP4 mouse (C) Channelrhodopsin evoked currents in a DRN GFP+ neuron associated to fluorescent and infra-red images (D) Schematics of ovBNST and CeA slices and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (E) Confocal images of patched neurons filled with biocytin and PKC-δ immunostaining in ovBNST and CeA. (F) Excitatory post-synaptic evoked currents recorded in ovBNST and CeA neurons upon optogenetic stimulation of DRN VIP fibers. Purple curves show inward currents abolished by application NMDA antagonists, APV and CNQX (black curve). (G) Donut graphs showing the proportion of neurons responding to optogenetic stimulation of DRN VIP fibers colocalizing with PKC-δ. (H) Ex vivo electrophysiological characterization of ovBNST and CeA excitatory post-synaptic currents evoked by optogenetic stimulation of DRN VIP fibers. Abbreviations: DRN: dorsal raphe nucleus; ovBNST oval nucleus of the bed nucleus of the stria terminalis; Aq: aqueduct of Sylvius; CeA: central nucleus of the amygdala; PPR: paired-pulse ratio.

Journal: bioRxiv

Article Title: A subset of dorsal raphe dopamine neurons is critical for survival-oriented vigilance

doi: 10.1101/2025.03.31.646345

Figure Lengend Snippet: (A) Schematics of a DRN slice and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (B) Confocal images of a patched GFP+ neuron filled with biocytin in the DRN of a Vip-Cre/COP4 mouse (C) Channelrhodopsin evoked currents in a DRN GFP+ neuron associated to fluorescent and infra-red images (D) Schematics of ovBNST and CeA slices and positioning of recording electrode and optic fiber in Vip-Cre/COP4 mice. (E) Confocal images of patched neurons filled with biocytin and PKC-δ immunostaining in ovBNST and CeA. (F) Excitatory post-synaptic evoked currents recorded in ovBNST and CeA neurons upon optogenetic stimulation of DRN VIP fibers. Purple curves show inward currents abolished by application NMDA antagonists, APV and CNQX (black curve). (G) Donut graphs showing the proportion of neurons responding to optogenetic stimulation of DRN VIP fibers colocalizing with PKC-δ. (H) Ex vivo electrophysiological characterization of ovBNST and CeA excitatory post-synaptic currents evoked by optogenetic stimulation of DRN VIP fibers. Abbreviations: DRN: dorsal raphe nucleus; ovBNST oval nucleus of the bed nucleus of the stria terminalis; Aq: aqueduct of Sylvius; CeA: central nucleus of the amygdala; PPR: paired-pulse ratio.

Article Snippet: Recordings of ovBNST neurons were made using patch electrodes made from borosilicate glass capillaries (GC150F10; Phymep) on a horizontal puller (Model P97; Sutter instruments) to obtain a pipette resistance of 5-8MΩ.

Techniques: Immunostaining, Ex Vivo