Journal: iScience

Article Title: The SNARE protein SNAP-25 is required for normal exocytosis at auditory hair cell ribbon synapses

doi: 10.1016/j.isci.2022.105628

Figure Lengend Snippet:

Article Snippet: AAV8.CMV.HI.eGFPCre.WPRE.SV40 , Addgene , Cat#105545.

Techniques: Recombinant, Electron Microscopy, Multiplex Assay, Software, Imaging, Injection

Journal: eLife

Article Title: Cortico-striatal action control inherent of opponent cognitive-motivational styles

doi: 10.7554/elife.100988

Figure Lengend Snippet: Figure 8. Timeline of experimental procedures and effects of clozapine N-oxide (CNO) on Cued-Triggered Turning Task (CTTT) performance. (a) Following initial handling and screening to identify the rat’s phenotype (17 goal trackers [GTs], 8 females; 12 sign-trackers [STs], 6 females; Figure 2), rats were infused with either a Cre-dependent inhibitory DREADD or the empty control vector (both expressing mCherry; b) into the lower layers of the prelimbic cortex, and a retrogradely transported, Cre-expressing plasmid into the dorsomedial striatum (DMS) (expressing eGFP). These rats continued to undergo CTTT performance training, followed either by (c) a test of the effects of vehicle or CNO on CTTT performance (the blue syringes symbolizing CNO administration) or by (d) implantation of an microelectrode array for the measurement of glutamate concentrations in the DMS and following vehicle or CNO administration. Following recovery from that surgery, the effects of CNO and vehicle on performance and performance- associated glutamate concentrations were assessed. In GTs, administration of CNO significantly reduced the relative number of cued turns (e), but not cued stops (f; e and f show individual data, mean and 95% CI; post hoc multiple comparisons: ***: p < 0.001, Tukey’s Honest Significant Difference test). CNO had no effects on cued turns or stops in rats expressing an empty control vector (see Results). In GTs, CNO administration attenuated cue-locked maximum peak glutamate concentrations during (residual) cued turns (g) and decreased the number of traces with just one glutamate peak during the cue period while increasing the frequency of two peaks during this period (h). Following the administration of CNO, residual turns took significantly more time to be initiated in GTs when compared with vehicle-treated GTs and CNO-treated STs (i). When followed by failures to turn, or misses, turn cue-locked maximum peak concentrations were not affected in GTs but reduced in STs (significant interaction between the effects of phenotype and CNO; j). Maximum peak glutamate concentrations locked to reward delivery again were higher in STs than in GTs, and CNO reduced these concentrations in both phenotypes (main effect of CNO, no interaction k); (multiple comparisons: **, ***: p < 0.01, 0.001).

Article Snippet: DOI: https://doi.org/10.7554/eLife.100988 24 of 32 Cre- dependent control plasmid pAAV- hSyn- DIO- mCherry (AddGene #50459- AAV8; titer of 2.2 × 1013 GC/ml), or the retrogradely transported Cre- expressing plasmid pENN- rAAV- hSyn- HI- eGFPCre- WPRE- SV40 (AddGene #105540- AAVrg; titer of 1.9 × 1013 GC/ml) were infused into the lower layers of the prelimbic cortex (for the anatomical organization of fronto- striatal projections see, e.g., Mailly et al., 2013) and the DMS, respectively.

Techniques: Control, Plasmid Preparation, Expressing, Microelectrode Array

Journal: eLife

Article Title: Cortico-striatal action control inherent of opponent cognitive-motivational styles

doi: 10.7554/elife.100988

Figure Lengend Snippet: Figure 9. Expression of eGFP (green channel), indicating the expression of the retrograde Cre-expressing vector in the dorsomedial striatum (DMS) and, following retrograde transport, in the medial frontal cortex. The presence of the amplified mCherry fluorescent reporter signal (red channel) in cortex indicated the expression of the inhibitory hM4Di DREADD vector. Illustration of expression efficacy ratings for eGFP in the striatum (a, b) and of mCherry in the prelimbic cortex (c, d), with a top score of 5 indicating complete or near-complete expression exclusively in the DMS projection field of prelimbic cortex efferent neurons (Mailly et al., 2013) and prelimbic cortex, respectively. The example of eGFP expression in the DMS in (e) (cc, corpus callosum) received a score of 2 (illustrated in a) because the expression field was located in part lateral to the prelimbic cortex projection field. (f) depicts an example of mCherry expression in the prelimbic cortex that was assigned a score of 4 as it was restricted to the deeper layers in the prelimbic cortex and extended dorsally into cingulate cortex. In goal trackers (GTs), but not sign-trackers (STs), mCherry expression scores in cortex were significantly correlated with the efficacy of clozapine N-oxide (CNO) to reduce cued turns (g; note that the ordinate in (g) depicts the difference between the proportion of turns in vehicle- minus CNO-treated rats, so that higher scores indicate greater CNO effects, and therefore higher expression scores were correlated with greater CNO effects). (h) depicts the retrogradely transported eGFP and the expression of mCherry, including the co-expressing of both fluorochromes, primarily in layer 5 of the prelimbic cortex. The brightened rectangular region in (h) is enlarged in (i) and shows neurons co-expressing nucleolar eGFP and cytoplasmic mCherry (white arrows in i). In GTs, but not STs, the proportion of double-labeled neurons in prelimbic cortex was significantly correlated with the efficacy of CNO to attenuate cued turns (j) note that higher scores on the ordinate depicts greater CNO effects.

Article Snippet: DOI: https://doi.org/10.7554/eLife.100988 24 of 32 Cre- dependent control plasmid pAAV- hSyn- DIO- mCherry (AddGene #50459- AAV8; titer of 2.2 × 1013 GC/ml), or the retrogradely transported Cre- expressing plasmid pENN- rAAV- hSyn- HI- eGFPCre- WPRE- SV40 (AddGene #105540- AAVrg; titer of 1.9 × 1013 GC/ml) were infused into the lower layers of the prelimbic cortex (for the anatomical organization of fronto- striatal projections see, e.g., Mailly et al., 2013) and the DMS, respectively.

Techniques: Expressing, Plasmid Preparation, Amplification, Labeling