Netrin-1 regulates the balance of synaptic glutamate signaling in the adult ventral tegmental area
- Department of Psychiatry and Behavioral Sciences, University of Washington, United States
- Molecular and Cellular Biology Program, University of Washington, United States
- Department of Pharmacology, University of Washington, United States
Figures
Figure 1
Netrin-1 is present in the adult ventral tegmental area (VTA) and expressed by both dopamine and GABA neurons.
3D display of Ntn1 (A) and Slc6a3 (B, dopamine marker) from the Allen Brain Atlas. (C–D) 20 X magnification images of in situ hybridization (RNAScope) for Ntn1 (green) and Slc32a1 (GABA marker; red, …
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Figure 1—source data 1
Cell counts.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig1-data1-v1.xlsx
Figure 2
Virally delivered CRISPR-Cas9 complex targeting the Ntn1 locus results in a significant reduction in Ntn1 antibody staining.
(A–B) Schematics summarizing cell type-specific knockout procedure. (A) Adult mice were injected bilaterally into the VTA with AAV-FLEX-SaCas9-HA-sgNtn1 and AAV-FLEX-YFP. Control mice received an …
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Figure 2—source data 1
Cell counts.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig2-data1-v1.xlsx
Figure 3 with 3 supplements
Loss of Ntn1 results in a significant reduction in excitatory postsynaptic current.
(A) Schematic of DAT-Cre dopamine specific Ntn1cKO. (B) Sample traces from control (top panel) and DAT Ntn1 cKO mice (bottom panel). (C–D) mEPSC amplitude (C) and frequency (D) measured from …
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Figure 3—source data 1
EPSCs and IPSCs from targeted cells.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig3-data1-v1.xlsx
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Figure 3—source data 2
Additional EPSC and IPSC data from non-targeted cells.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig3-data2-v1.xlsx
Figure 3—figure supplement 1
No significant differences in inhibitory synaptic connectivity associated with Ntn1 loss of function.
(A) mIPSC amplitude and frequency of DAT-Cre fluorescently identified dopamine neurons (n=20 controls, n=27 cKO, t=0.8283, df = 45 p>0.05 and t=0.6668, df = 45 p>0.05). (B) mIPSC amplitude and …
Figure 3—figure supplement 2
No significant differences in excitatory or inhibitory synaptic connectivity in non-targeted cell types.
(A) Miniature excitatory postsynaptic current (mEPSC) amplitude and frequency recorded from non-fluorescent cells in DAT-Cre mice (presumptively non-dopamine neurons) (n=16 controls, n=11 cKO, …
Figure 3—figure supplement 3
Loss of Netrin function results in significant decrease in AMPA response.
Analysis of 1/CV2 from miniature excitatory postsynaptic current (mEPSC) data in (A) DAT-Cre mice measured from fluorescently identified dopamine neurons (n=35 controls, n=33 cKO, t=1.252, df = 66. …
Figure 4 with 1 supplement
Ntn1 cKO in DA neurons results in little behavioral alteration.
(A) Schematic summarizing cell type-specific knockout procedure. (B) Distance traveled in 15 min bins over the course of three nights and two days (n=21 control; n=15 cKO, Two-way ANOVA, Group F(1, …
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Figure 4—source data 1
Behavioral data for Figure 4.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
Additional behavioral analysis of Ntn1 cKO DAT-Cre mice.
(A) Average ambulatory activity (beam breaks) averaged across three nights and two days (n=21 control; n=18 cKO, Night: t=0.3938, df = 37, p=0.6960, Day: t=1.271, df = 37, p=0.2117). (B) Distance …
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Figure 4—figure supplement 1—source data 1
Behavioral data for Figure 4—figure supplement 1.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig4-figsupp1-data1-v1.xlsx
Figure 5 with 1 supplement
Ntn1 cKO in GABA ventral tegmental area (VTA) neurons resulted in significant behavioral alterations.
(A) Schematic summarizing cell type-specific knockout procedure. (B) Distance traveled in 15 min bins over the course of three nights and two days (n=26 controls, n=23 cKO, Two-way ANOVA Group F(1, …
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Figure 5—source data 1
Behavioral data for Figure 5.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Additional behavioral analysis of Ntn1 cKO Vgat-Cre mice.
(A) Average ambulatory activity (beam breaks) averaged across three nights and two days (n=26 control; n=26 cKO, t=3.109, df = 50 **p=0.01 and t=3.227, df = 50 **p=0.01). (B) Distance traveled …
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Figure 5—figure supplement 1—source data 1
Behavioral data for Figure 5—figure supplement 1.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig5-figsupp1-data1-v1.xlsx
Figure 6 with 1 supplement
Ntn1 cKO in DATIRES::Vgat-Cre mice partially rescues behavioral phenotype.
(A) Model of Ntn1 loss of function in the ventral tegmental area (VTA) on excitatory and inhibitory balance. (B) Schematic of GABA and Dopamine Ntn1 cKO. (C) Distance traveled in 15 min bins over …
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Figure 6—source data 1
Behavioral data for Figure 6.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Average day and night locomotion in DAT-Cre::Vgat-Cre mice.
(A) Ambulatory activity (beam breaks) averaged across three nights and two days (n=26 controls n=21 cKO, Night: t=1.010, df = 45, p=0.3178 and Day: t=2.091, df = 45 *p<0.05). (B) Distance traveled …
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Figure 6—figure supplement 1—source data 1
Behavioral data for Figure 6—figure supplement 1.
- https://cdn.elifesciences.org/articles/83760/elife-83760-fig6-figsupp1-data1-v1.xlsx
