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. 2019 Sep 12;10(1):4140.
doi: 10.1038/s41467-019-12122-8.

A Novel Role for the Actin-Binding Protein Drebrin in Regulating Opiate Addiction

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Free PMC article

A Novel Role for the Actin-Binding Protein Drebrin in Regulating Opiate Addiction

Jennifer A Martin et al. Nat Commun. .
Free PMC article

Abstract

Persistent transcriptional and morphological events in the nucleus accumbens (NAc) and other brain reward regions contribute to the long-lasting behavioral adaptations that characterize drug addiction. Opiate exposure reduces the density of dendritic spines on medium spiny neurons of the NAc; however, the underlying transcriptional and cellular events mediating this remain unknown. We show that heroin self-administration negatively regulates the actin-binding protein drebrin in the NAc. Using virus-mediated gene transfer, we show that drebrin overexpression in the NAc is sufficient to decrease drug seeking and increase dendritic spine density, whereas drebrin knockdown potentiates these effects. We demonstrate that drebrin is transcriptionally repressed by the histone modifier HDAC2, which is relieved by pharmacological inhibition of histone deacetylases. Importantly, we demonstrate that heroin-induced adaptations occur only in the D1+ subset of medium spiny neurons. These findings establish an essential role for drebrin, and upstream transcriptional regulator HDAC2, in opiate-induced plasticity in the NAc.

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Drebrin mediates behavioral plasticity in the NAc. a Mean numbers of infusions of saline (gray) or heroin (blue) self-administered over 10 days (two-way repeated-measures ANOVA: drug effect, F1,41 = 76.74, P < 0.001; day effect, F9,369 = 7.899, P < 0.001; interaction, F9,369 = 8.882, P < 0.001, n= 19–24 per group). Actin cycling (Student’s t-test: t9 = 2.632, P = 0.027, n= 6–7 per group) (b) and drebrin protein levels (Student’s t-test: t13 = 2.953, P= 0.011, n= 7–8 per group) (c) in the NAc following heroin SA. Within-session heroin SA dose response (μg kg−1 infusion−1 [inf]) following intra-NAc injections of HSV-drebrin or HSV-GFP (two-way ANOVA: dose effect, F3,72 = 4.314, P < 0.001; virus effect, F1,72 = 10.68, P < 0.01; interaction, F3,71 = 0.56, P> 0.05, n= 10–11 per group) (d) and HSV-CRISPR-Cas9 drebrin (CRISPR-drebrin) or HSV-scrambled (two-way ANOVA: dose effect, F3,60 = 8.796, P < 0.001; virus effect, F1,60 = 4.467, P < 0.05; interaction, F3,60 = 1.864, P> 0.05, n= 9–8 per group) (e). Total active responses during drug-induced reinstatement following virus-mediated overexpression of drebrin (two-way ANOVA: drug effect, F1,18 = 23.68, P < 0.001; virus effect, F1,18 = 3.782, P= 0.068; interaction, F1,18 = 6.63, P= 0.019; saline-GFP vs heroin-GFP, P= 0.003; heroin-GFP vs heroin-drebrin, P= 0.017; saline-GFP vs heroin-drebrin, P= 0.201; n= 5–6 per group) (f) or CRISPR-drebrin (two-way ANOVA: drug effect, F1,31 = 53.71, P < 0.001; virus effect, F1,31 = 5.937, P < 0.05; interaction, F1,31 = 7.17, P= 0.01, saline-scrambled vs heroin-scrambled, P = 0.02; heroin-scrambled vs heroin-CRISPR, P = 0.003; n= 6–10 per group) (g) in the NAc. Data are expressed as means ± SEMs. *P < 0.05, **P < 0.01
Fig. 2
Fig. 2
Epigenetic regulation of drebrin expression following heroin SA. HDAC2 protein expression (Student’s t-test: t12 = 2.279, P < 0.05, n= 7 per group) (a) and binding along AP-1 (Student’s t-test: t14 = 2.435, P < 0.05, n= 8 per group) and SBE (Student’s t-test: t14 = 2.17, P < 0.05, n= 8 per group) sites along the drebrin promoter (b) following 10 days of heroin SA. HDAC2 binding along AP-1 (Student’s t-test: t6 = 3.383, P < 0.05, n= 4 per group) and SBE (Student’s t-test: t6 = 31.313, P= 0.23, n= 4 per group) sites on the drebrin promoter (c), drebrin mRNA levels (Student’s t-test: t13 = 2.056, P < 0.05, n= 7–8 per group) (d), and total active responses (Student’s t-test: t20 = 2.329, P < 0.05, n= 11 per group) (e) following HDAC2 inhibition via MI-192 microinjections. Data are expressed as means ± SEMs. *P < 0.05
Fig. 3
Fig. 3
Drebrin in the NAc regulates opiate-induced structural plasticity. a Spine densities on MSNs in the NAc following intra-NAc HSV-drebrin expression (two-way ANOVA: drug effect, F2,15 = 0.482, P < 0.05; virus effect, F1,15 = 13.71, P> 0.05; interaction, F2,15 = 5.91, P < 0.01, saline-GFP vs heroin-GFP, P= 0.008; heroin-GFP vs heroin-drebrin, P= 0.0002; saline-GFP vs heroin-drebrin, P= 0.125; n = 5–6 per group). b Spine densities on MSNs in the NAc following intra-NAc CRISPR-Cas9 drebrin (two-way ANOVA: drug effect, F1,10 = 20.92, P < 0.001; virus effect, F1,10 = 3.858, P> 0.05; interaction, F1,10 = 2.138, P> 0.05; saline-scrambled vs heroin-scrambled, P = 0.05; heroin-scrambled vs heroin-CRISPR, P = 0.04; n = 3–4 per group). c Representative images of virus-infected neurons; scale bars, 5 μm. Data are expressed as means ± SEMs. *P < 0.05, ***P < 0.001
Fig. 4
Fig. 4
Drebrin restores opiate-induced deficits in synaptic plasticity. a Intra-NAc expression of HSV-drebrin rescues morphine-induced changes in F/G-actin ratios (two-way ANOVA: drug effect, F1,28 = 0.979, P > 0.05; virus effect, F1,28 = 45.14, P < 0.01; interaction, F1,28 = 10.62, P < 0.01; saline-GFP vs morphine-GFP, P = 0.005; morphine-GFP vs morphine-drebrin, P < 0.0001; saline-GFP vs morphine-drebrin, P = 0.0004; saline-GFP vs saline-drebrin, P = 0.02; n = 8 per group). Surface protein expression of AMPA receptor subunit GluA1 (Student’s t-test: t10 = 1.949, P < 0.05, n = 6 per group) (b) and NMDA receptor subunits NR2B (Student’s t-test: t13 = 2.856, P < 0.05, n = 7–8 per group) (c) and NR1 (Student’s t-test: t10 = 2.405, P < 0.05, n = 6 per group) (d) in the NAc following heroin SA. Data are expressed as means ± SEMs. *P < 0.05
Fig. 5
Fig. 5
Cell-type-specific expression of drebrin alters behavioral and structural plasticity. a Drebrin mRNA expression in D1-expressing (Student’s t-test: t10 = 1.864, P < 0.05, n = 5–7 per group) and D2-expressing (Student’s t-test: t10 = 0.82, P > 0.05, n = 5–7 per group) MSNs following heroin SA. Total active responses following cell-type-specific overexpression of drebrin in D1-expressing (Student’s t-test: t10 = 3.153, P < 0.05, n = 6 per group) (b) or D2-expressing (Student’s t-test: t7 = 0.1394, P > 0.05, n = 4–5 per group) (c) MSNs. Dendritic spine densities following cell-type-specific overexpression of drebrin in D1-expressing (Student’s t-test: t9 = 1.914, P < 0.05, n = 5–6 per group) (d) or D2-expressing (Student’s t-test: t7 = 0.537, P > 0.05, n = 4–5 per group) (e) MSNs. (f) Representative images of virus-infected neurons; scale bars, 5 μm. Data are expressed as means ± SEMs. *P < 0.05

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