Journal: eLife

Article Title: Optical dopamine monitoring with dLight1 reveals mesolimbic phenotypes in a mouse model of neurofibromatosis type 1

doi: 10.7554/eLife.48983

Figure Lengend Snippet: ( A ) Average ( left ) and trial-by-trial ( right ) fluorescent dopamine response to a 10 s overhead light stimulus. ( B ) Average ( left ) and trial-by-trial ( right ) fluorescent dopamine response to a 10 s auditory stimulus (5 kHz tone). ( C ) Nf1 +/- mice had greater peak responses to light (p<0.001) but not tone onset (p>0.99) compared to Nf1 +/+ mice (n +/+ = 10, n +/- = 10; two-way ANOVA with Bonferroni post hoc tests; F 1,36 = 7.27, p genotype x condition = 0.01; F 1,36 = 15.48, p genotype <0.001). In both genotypes, responses to light were greater than responses to tone (F 1,36 = 71.02, p stimulus <0.001; p +/+ = 0.002, p +/- < 0.001). ( D ) No difference in cued fear conditioning was observed when a tone-only CS was used (n +/+ = 9, n +/- = 10; two-way repeated measures ANOVA; F 9,153 = 0.26, p genotype x trial = 0.98). ( E ) Nf1 +/- mice exhibited increased freezing ( left ) and decreased latency to freeze ( right ) in tone-only CS trials (n = 10) compared to light and tone (n = 18; unpaired t-test; freezing: t 26 = 3.75, p<0.001; latency: t 26 = 3.75, p<0.00) or interleaved light trials (n = 10; paired t-test; freezing: t 9 = 5.30, p<0.001; latency: t 9 = 3.48, p=0.007). No differences in freezing or latency to freeze was observed between tone-only CS trials (n = 9) and light and tone (n = 13; unpaired t-test; freezing: t 20 = 1.66, p=0.11; latency: t 20 = 0.81, p=0.43) or interleaved light trials (n = 10; paired t-test; freezing: t 8 = 0.42, p=0.69; latency: t 8 = 1.49, p=0.19) in Nf1 +/+ mice. ( F ) Nf1 +/+ (n = 17) and Nf1 +/- mice (n = 23) had similar reaction times to a looming stimulus ( left ; t 38 = 0.79, p=0.43), yet Nf1 +/+ mice were more likely to escape to the shelter after stimulus presentation ( left ) and exhibited shorter latency to the first freezing episode after looming onset than Nf1 +/- mice (t 38 = 3.24, p=0.003). ( G ) Optogenetic stimulation of the ventral superior colliculus (vSC) produced time-locked dopamine release in the LNAc (n = 3 mice; average trace, left ; trial-by-trial response, right ). ( H ) Representative confocal image showing tyrosine hydroxylase (TH)-positive dopaminergic and Th -Off-NpHR-eYFP neurons in the VTA (scale: 50 μm). ( I ) In the absence of photoinhibition, VTA Th -Off-NpHR-eYFP Nf1 +/- mice (n = 12) were more likely to escape to the shelter ( left ) and had shorter latency to the first freezing episode ( right ; unpaired t-test; t 22 = 2.36, p=0.03) compared with VTA Th -Off-mCherry Nf1 +/+ mice (n = 12). Optogenetic inhibition of VTA non-Th neurons with 532 nm light (5 mW, 30 Hz, 20 ms pulse width) decreased the probability of escape to the shelter ( left ) and increased the latency to the first freezing episode ( right ; paired t-test; t 11 = 3.82, p=0.003) in VTA Th -Off-NpHR-eYFP Nf1 +/- mice to levels that were similar to VTA Th -Off-mCherry Nf1 +/+ mice (unpaired t-test; Nf1 +/- Laser On vs Nf1 +/+ Laser Off : t 22 = 0.98, p=0.34; Nf1 +/- Laser On vs Nf1 +/+ Laser On : t 22 = 1.09, p=0.29). No difference was observed in VTA Th -Off-mCherry Nf1 +/+ mice between stimulation conditions (paired t-test; t 11 = 0.02, p=0.99). *denotes p<0.05. # denotes p<0.05 vs light stimulus (panel C). Data presented as mean ± SEM.

Article Snippet: For optogenetic inhibition of VTA non-Th neurons, 500 nL of a 1:4 mixture of AAV9- Th -PI-Cre-SV40 (gift of James M. Wilson, Addgene viral prep # 107788-AAV9) and AAV-DJ-Ef1α-DO-eNpHR3.0-eYFP-WPRE-pA (gift of Bernardo Sabatini, Addgene # 37087) or AAV-DJ-Ef1α-DO-mCherry-WPRE-pA (gift of Bernardo Sabatini, Addgene # 37119) was injected bilaterally into the VTA (antero-posterior: −3.3 mm, medio-lateral: ± 0.5 mm, dorso-ventral: −4.2 mm relative to Bregma), followed by implantation of 5 mm long, 300 μm mono fiber-optic cannulae (MFC_300/330–0.48_5 mm_ZF1.25_FLT; antero-posterior: −3.3 mm, medio-lateral: ± 1.84 mm, dorso-ventral: −3.59 mm relative to Bregma) at angle of twenty degrees.

Techniques: Produced, Inhibition

Journal: eLife

Article Title: Optical dopamine monitoring with dLight1 reveals mesolimbic phenotypes in a mouse model of neurofibromatosis type 1

doi: 10.7554/eLife.48983

Figure Lengend Snippet:

Article Snippet: For optogenetic inhibition of VTA non-Th neurons, 500 nL of a 1:4 mixture of AAV9- Th -PI-Cre-SV40 (gift of James M. Wilson, Addgene viral prep # 107788-AAV9) and AAV-DJ-Ef1α-DO-eNpHR3.0-eYFP-WPRE-pA (gift of Bernardo Sabatini, Addgene # 37087) or AAV-DJ-Ef1α-DO-mCherry-WPRE-pA (gift of Bernardo Sabatini, Addgene # 37119) was injected bilaterally into the VTA (antero-posterior: −3.3 mm, medio-lateral: ± 0.5 mm, dorso-ventral: −4.2 mm relative to Bregma), followed by implantation of 5 mm long, 300 μm mono fiber-optic cannulae (MFC_300/330–0.48_5 mm_ZF1.25_FLT; antero-posterior: −3.3 mm, medio-lateral: ± 1.84 mm, dorso-ventral: −3.59 mm relative to Bregma) at angle of twenty degrees.

Techniques: Recombinant, Plasmid Preparation, Produced, Software, Control, Refractive Index