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Basal forebrain projections to the lateral habenula modulate aggression reward

View Article: PubMed Central - PubMed

ABSTRACT

Maladaptive aggressive behavior is associated with a number of neuropsychiatric disorders1 and is thought to partly result from inappropriate activation of brain reward systems in response to aggressive or violent social stimuli2. Nuclei within the ventromedial hypothalamus3–5, extended amygdala6 and limbic7 circuits are known to encode initiation of aggression; however, little is known about the neural mechanisms that directly modulate the motivational component of aggressive behavior8. To address this, we established a mouse model to measure the valence of aggressive inter-male social interaction with a smaller subordinate intruder as reinforcement for the development of conditioned place preference (CPP). Aggressors (AGG) develop a CPP, while non-aggressors (NON) develop a conditioned place aversion (CPA), to the intruder-paired context. Further, we identify a functional GABAergic projection from the basal forebrain (BF) to the lateral habenula (lHb) that bi-directionally controls the valence of aggressive interactions. Circuit-specific silencing of GABAergic BF-lHb terminals of AGG with halorhodopsin (NpHR3.0) increases lHb neuronal firing and abolishes CPP to the intruder-paired context. Activation of GABAergic BF-lHb terminals of NON with channelrhodopsin (ChR2) decreases lHb neuronal firing and promotes CPP to the intruder-paired context. Lastly, we show that altering inhibitory transmission at BF-lHb terminals does not control the initiation of aggressive behavior. These results demonstrate that the BF-lHb circuit plays a critical role in regulating the valence of inter-male aggressive behavior and provide novel mechanistic insight into the neural circuits modulating aggression reward processing.

No MeSH data available.


Multiunit anesthetized optrode recordings(a) Schematic of in vivo anesthetized multi-unit optrode recording procedure (left) and representative optrode placement in lHb (right; scale bar = 200 μm). Heatmaps of normalized firing rates for lHb neurons in response to BF terminal stimulation with (b) ChR2BF→lHb or (c) NpHR3BF→lHb and averaged spike wave-form shown below for pre-stimulation, stimulation and post-stimulation epochs. (e) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with ChR2BF→lHb for all identified cells (F2,134 = 8.249, one-way repeated-measure ANOVA P < 0.001; post hoc test, *P < 0.05; n = 68 cells from 3 mice) and cells that significantly decreased firing during the stimulation epoch (F7,105 = 8.868, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 16/68 cells from 3 mice). (f) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with NpHR3BF→lHb for all identified cells (F2,128 = 10.32, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 65/65 cells from 3 mice) and cells that significantly increased firing during the stimulation epoch (F7,203 = 17.58, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 30/65 cells from 3 mice). BF, basal forebrain; lHb, lateral habenula; mHb, medial habenula; DAPI, 4′,6-diamidino-2-phenylindole. Summary data are represented as mean ± s.e.m.
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Figure 9: Multiunit anesthetized optrode recordings(a) Schematic of in vivo anesthetized multi-unit optrode recording procedure (left) and representative optrode placement in lHb (right; scale bar = 200 μm). Heatmaps of normalized firing rates for lHb neurons in response to BF terminal stimulation with (b) ChR2BF→lHb or (c) NpHR3BF→lHb and averaged spike wave-form shown below for pre-stimulation, stimulation and post-stimulation epochs. (e) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with ChR2BF→lHb for all identified cells (F2,134 = 8.249, one-way repeated-measure ANOVA P < 0.001; post hoc test, *P < 0.05; n = 68 cells from 3 mice) and cells that significantly decreased firing during the stimulation epoch (F7,105 = 8.868, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 16/68 cells from 3 mice). (f) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with NpHR3BF→lHb for all identified cells (F2,128 = 10.32, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 65/65 cells from 3 mice) and cells that significantly increased firing during the stimulation epoch (F7,203 = 17.58, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 30/65 cells from 3 mice). BF, basal forebrain; lHb, lateral habenula; mHb, medial habenula; DAPI, 4′,6-diamidino-2-phenylindole. Summary data are represented as mean ± s.e.m.

Mentions: To determine the functional contribution of BF-lHb projections, we conducted optogenetic circuit-specific terminal photostimulation in combination with slice electrophysiology with channelrhodopsin (AAV2-hSyn-ChR2(H134R)-eYFP) or halorhodopsin (AAV2-hSyn-NpHR3.0-eYFP), identifying photostimulation parameters that produce robust transient lHb activation or inhibition without rebound neuronal firing. ChR2BF→lHb terminal photostimulation with 40 Hz resulted in significantly decreased lHb firing rates (Fig. 2j–k), while NpHR3BF→lHb (8-s on, 2-s off) terminal photostimulation resulted in a robust increase in postsynaptic lHb firing rates (Fig. 2l–m). Importantly, whole cell recordings from lHb neurons during ChR2BF→lHb terminal photostimulation showed a significant increase in inhibitory postsynaptic currents (IPSCs) that was completely blocked by the GABAA receptor antagonist, gabazine (Extended Data Figure 4d–e). Optically induced IPSCs exhibited a response delay of ~7 ms (Extended Data Figure 4f), which is in line with previously published response delays for ChR2 at monosynaptic circuits. Similarly, anterograde tracing of BF terminals in the lHb revealed that they were co-localized with vesicular GABA transporter (VGAT), but not vesicular glutamate transporter 1 (VGLUT1) (Extended Data Figure 4g). To validate these findings within an intact system, we utilized multi-electrode recording of postsynaptic lHb firing rates in anesthetized mice in combination with terminal photostimulation (Extended Data Figure 5a). Results show that activation (40 Hz ChR2BF→lHb), or inhibition (8-s on, 2-s off NpHR3BF→lHb) of presynaptic BF terminals in the lHb resulted in decreased or increased lHb postsynaptic neuronal firing, respectively (Extended Data Figure 5b–d). These functional in vitro and in vivo recordings of ChR2BF→lHb and NpHR3BF→lHb, confirm inhibitory GABAergic control over circuit activity and demonstrate reliable temporal control of lHb firing rates by optogenetic tools for in vivo behavioral analysis.


Basal forebrain projections to the lateral habenula modulate aggression reward
Multiunit anesthetized optrode recordings(a) Schematic of in vivo anesthetized multi-unit optrode recording procedure (left) and representative optrode placement in lHb (right; scale bar = 200 μm). Heatmaps of normalized firing rates for lHb neurons in response to BF terminal stimulation with (b) ChR2BF→lHb or (c) NpHR3BF→lHb and averaged spike wave-form shown below for pre-stimulation, stimulation and post-stimulation epochs. (e) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with ChR2BF→lHb for all identified cells (F2,134 = 8.249, one-way repeated-measure ANOVA P < 0.001; post hoc test, *P < 0.05; n = 68 cells from 3 mice) and cells that significantly decreased firing during the stimulation epoch (F7,105 = 8.868, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 16/68 cells from 3 mice). (f) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with NpHR3BF→lHb for all identified cells (F2,128 = 10.32, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 65/65 cells from 3 mice) and cells that significantly increased firing during the stimulation epoch (F7,203 = 17.58, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 30/65 cells from 3 mice). BF, basal forebrain; lHb, lateral habenula; mHb, medial habenula; DAPI, 4′,6-diamidino-2-phenylindole. Summary data are represented as mean ± s.e.m.
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Figure 9: Multiunit anesthetized optrode recordings(a) Schematic of in vivo anesthetized multi-unit optrode recording procedure (left) and representative optrode placement in lHb (right; scale bar = 200 μm). Heatmaps of normalized firing rates for lHb neurons in response to BF terminal stimulation with (b) ChR2BF→lHb or (c) NpHR3BF→lHb and averaged spike wave-form shown below for pre-stimulation, stimulation and post-stimulation epochs. (e) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with ChR2BF→lHb for all identified cells (F2,134 = 8.249, one-way repeated-measure ANOVA P < 0.001; post hoc test, *P < 0.05; n = 68 cells from 3 mice) and cells that significantly decreased firing during the stimulation epoch (F7,105 = 8.868, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 16/68 cells from 3 mice). (f) Percent of cells by firing response (top) and average normalized lHb firing rate (bottom) after BF-lHb terminal stimulation with NpHR3BF→lHb for all identified cells (F2,128 = 10.32, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 65/65 cells from 3 mice) and cells that significantly increased firing during the stimulation epoch (F7,203 = 17.58, one-way repeated-measure ANOVA P < 0.0001; post hoc test, *P < 0.05; n = 30/65 cells from 3 mice). BF, basal forebrain; lHb, lateral habenula; mHb, medial habenula; DAPI, 4′,6-diamidino-2-phenylindole. Summary data are represented as mean ± s.e.m.
Mentions: To determine the functional contribution of BF-lHb projections, we conducted optogenetic circuit-specific terminal photostimulation in combination with slice electrophysiology with channelrhodopsin (AAV2-hSyn-ChR2(H134R)-eYFP) or halorhodopsin (AAV2-hSyn-NpHR3.0-eYFP), identifying photostimulation parameters that produce robust transient lHb activation or inhibition without rebound neuronal firing. ChR2BF→lHb terminal photostimulation with 40 Hz resulted in significantly decreased lHb firing rates (Fig. 2j–k), while NpHR3BF→lHb (8-s on, 2-s off) terminal photostimulation resulted in a robust increase in postsynaptic lHb firing rates (Fig. 2l–m). Importantly, whole cell recordings from lHb neurons during ChR2BF→lHb terminal photostimulation showed a significant increase in inhibitory postsynaptic currents (IPSCs) that was completely blocked by the GABAA receptor antagonist, gabazine (Extended Data Figure 4d–e). Optically induced IPSCs exhibited a response delay of ~7 ms (Extended Data Figure 4f), which is in line with previously published response delays for ChR2 at monosynaptic circuits. Similarly, anterograde tracing of BF terminals in the lHb revealed that they were co-localized with vesicular GABA transporter (VGAT), but not vesicular glutamate transporter 1 (VGLUT1) (Extended Data Figure 4g). To validate these findings within an intact system, we utilized multi-electrode recording of postsynaptic lHb firing rates in anesthetized mice in combination with terminal photostimulation (Extended Data Figure 5a). Results show that activation (40 Hz ChR2BF→lHb), or inhibition (8-s on, 2-s off NpHR3BF→lHb) of presynaptic BF terminals in the lHb resulted in decreased or increased lHb postsynaptic neuronal firing, respectively (Extended Data Figure 5b–d). These functional in vitro and in vivo recordings of ChR2BF→lHb and NpHR3BF→lHb, confirm inhibitory GABAergic control over circuit activity and demonstrate reliable temporal control of lHb firing rates by optogenetic tools for in vivo behavioral analysis.

View Article: PubMed Central - PubMed

ABSTRACT

Maladaptive aggressive behavior is associated with a number of neuropsychiatric disorders1 and is thought to partly result from inappropriate activation of brain reward systems in response to aggressive or violent social stimuli2. Nuclei within the ventromedial hypothalamus3&ndash;5, extended amygdala6 and limbic7 circuits are known to encode initiation of aggression; however, little is known about the neural mechanisms that directly modulate the motivational component of aggressive behavior8. To address this, we established a mouse model to measure the valence of aggressive inter-male social interaction with a smaller subordinate intruder as reinforcement for the development of conditioned place preference (CPP). Aggressors (AGG) develop a CPP, while non-aggressors (NON) develop a conditioned place aversion (CPA), to the intruder-paired context. Further, we identify a functional GABAergic projection from the basal forebrain (BF) to the lateral habenula (lHb) that bi-directionally controls the valence of aggressive interactions. Circuit-specific silencing of GABAergic BF-lHb terminals of AGG with halorhodopsin (NpHR3.0) increases lHb neuronal firing and abolishes CPP to the intruder-paired context. Activation of GABAergic BF-lHb terminals of NON with channelrhodopsin (ChR2) decreases lHb neuronal firing and promotes CPP to the intruder-paired context. Lastly, we show that altering inhibitory transmission at BF-lHb terminals does not control the initiation of aggressive behavior. These results demonstrate that the BF-lHb circuit plays a critical role in regulating the valence of inter-male aggressive behavior and provide novel mechanistic insight into the neural circuits modulating aggression reward processing.

No MeSH data available.