Journal: Cell reports

Article Title: Opposing Regulation of Cocaine Seeking by Glutamate and GABA Neurons in the Ventral Pallidum

doi: 10.1016/j.celrep.2020.01.023

Figure Lengend Snippet: (A) Outline of the protocol used in the chemogenetic and miniscope Ca 2+ imaging experiments. (B) Representative example of non-selective Gq-DREADD expression in the VP. ac, anterior commissure; BST, bed nucleus of the stria terminalis; cc, corpus callosum; DBH, diagonal horizontal band of Broca; f, fornix; ic, interior commissure; IPAC, interstitial nucleus of the posterior anterior commissure; lv, lateral ventricle; POA, preoptic area; SI, substantia innominata. Scale bar, 500 um. mCitrine expression is pseudocolored red for clarity. Substance P (green) was used as a counterstain to outline the borders of the VP. (C) Cocaine self-administration and extinction in wild-type mice expressing Gq-DREADD in VP neurons. (D and E) Simultaneously stimulating all VP neurons prior to a late extinction session (Ext) (D) or cue seeking in mice transduced with Gq DREADD in VP neurons (E) increased cocaine seeking. Data are presented as mean ± SEM. *p < 0.05 comparing vehicle and CNO (active), +p < 0.05 comparing extinction and cue seeking (active), and #p < 0.05 comparing vehicle and CNO (inactive).

Article Snippet: Mice were injected with the Cre dependent genetically encoded Ca 2+ indicator gCaMP6f (AAV1-hSyn-DIO-gCaMP6f; titer: ~1*10 13 GC/ml; University of Pennsylvania vector core and Addgene) into the VP, followed by the implantation of a GRIN lens (7.3 mm by 0.6 mm; Inscopix) above the VP.

Techniques: Imaging, Expressing, Transduction

Journal: Cell reports

Article Title: Opposing Regulation of Cocaine Seeking by Glutamate and GABA Neurons in the Ventral Pallidum

doi: 10.1016/j.celrep.2020.01.023

Figure Lengend Snippet: (A) Representative micrograph of GRIN lens implantation site in a Vglut2-IRES-Cre mouse above VP to record from VP Glu neurons expressing gCaMP6f. Inset shows example frame from miniature microscope video. BST, bed nucleus of the stria terminalis; cc, corpus callosum; DBH, diagonal horizontal band of Broca; f, fornix; ic, interior commissure; IPAC, interstitial nucleus of the posterior anterior commissure; lv, lateral ventricle; POA, preoptic area; SI, substantia innominata. Scale bar, 700 μm. (B) Self-administration of cocaine (with dummy camera) and extinction training in all mice. (C) Active and inactive nose pokes made in all mice used for Ca 2+ imaging during Ext1, Ext10, and Reinst. * p < 0.05 comparing Ext10 to Ext1 and Reinst. (D) Representative traces showing calcium events recorded from VP GABA , VP Penk , and VP Glu neurons. Red dots indicate registered Ca 2+ events. (E) Comparisons between the average event rate across cell types and sessions. Comparisons between cell types revealed that VP Glu neurons display a significantly higher Ca 2+ event rate across all conditions compared with VP GABA and VP Penk neurons and that the VP GABA Ca 2+ event rate is elevated compared with VP Penk during Reinst. Comparisons within cell types showed that extinction learning increases the frequency of Ca 2+ events in VP Glu (Ext1 versus Ext10 and Reinst), while Reinst selectively increases the Ca 2+ event rate of VP GABA (Reinst versus Ext1 and Ext10). *p < 0.05 compared with cue seeking within cell group, +p < 0.05 compared with Ext1 within cell group, ^p < 0.05 compared with VP Glu neurons in the same session, and #p < 0.05 compared with VP Penk in the same session. (F) Cumulative spikes across the 2 h recording session. n = cell number per session over (mouse number). *p < 0.05 comparing between sessions. All data are presented as mean ± SEM.

Article Snippet: Mice were injected with the Cre dependent genetically encoded Ca 2+ indicator gCaMP6f (AAV1-hSyn-DIO-gCaMP6f; titer: ~1*10 13 GC/ml; University of Pennsylvania vector core and Addgene) into the VP, followed by the implantation of a GRIN lens (7.3 mm by 0.6 mm; Inscopix) above the VP.

Techniques: Expressing, Microscopy, Imaging

Journal: Cell reports

Article Title: Opposing Regulation of Cocaine Seeking by Glutamate and GABA Neurons in the Ventral Pallidum

doi: 10.1016/j.celrep.2020.01.023

Figure Lengend Snippet:

Article Snippet: Mice were injected with the Cre dependent genetically encoded Ca 2+ indicator gCaMP6f (AAV1-hSyn-DIO-gCaMP6f; titer: ~1*10 13 GC/ml; University of Pennsylvania vector core and Addgene) into the VP, followed by the implantation of a GRIN lens (7.3 mm by 0.6 mm; Inscopix) above the VP.

Techniques: Virus, Plasmid Preparation, Recombinant, RNAscope, Multiplex Assay, Software

Journal: Nature Communications

Article Title: Multiplexed lipid nanoparticle barcoding reveals tissue-dynamic kinetic insights and enriched cellular tropism in hepatic zones

doi: 10.1038/s41467-025-68103-7

Figure Lengend Snippet: a Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Liver SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase shows liver-specific protein production. b , c Cre qPCR performed on liver, lung, and spleen 1-h ( b ) or 24-h ( c ) after administration of Liver SORT LNPs at various doses. d , e Linear regression analysis comparing tdTom signal vs. 1-h qPCR ( d ) or 24-h qPCR ( e ) shows linear correlation that is modestly stronger at 1-h. f Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Lung SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase. g , h Cre qPCR performed on liver, lung, and spleen 1-h ( g ) or 24-h ( h ) after administration of Lung SORT LNPs at various doses. i , j Linear regression analysis comparing tdTom signal vs. 1-h qPCR ( i ) or 24-h qPCR ( j ) shows linear correlation that is modestly stronger at 1-h. k Endogenous Rosa-tdTom fluorescent micrographs of Ai14 mouse liver, lung, and spleen 24 h post-intravenous (IV) administration of Spleen SORT LNPs carrying 0.1, 0.25, or 1.0 mg/kg Cre recombinase shows spleen-specific protein production. l , m Cre qPCR performed on liver, lung, and spleen 1-h ( l ) or 24-h ( m ) after administration of Liver SORT LNPs at various doses. n , o Linear regression analysis comparing tdTom signal vs. 1-hour qPCR ( n ) or 24-h qPCR ( o ) shows linear correlation that is somewhat stronger at 1-h. qPCR data ( b – e , g – j , l – o ) are presented as the mean ± SEM of N = 3 mice per dose, per timepoint, per formulation; three wells per measurement. For tdTom quantification: mean ± SEM, N = 3 image fields. Correlation analyses ( d , e , i , j , n , o ) show Pearson’s R -squared with 95% CI calculated after confirming homoscedasticity. Scale bars ( a , f , k ): 200 µm.

Article Snippet: Barcodes and a PCR adapter were cloned into an in vitro transcription (IVT) vector encoding Cre recombinase with a DNA-encoded polyA tail using Q5 SDM kit according to the manufacturer's instructions (New England Biolabs, E0554S).

Techniques: Formulation

Journal: The Journal of Clinical Investigation

Article Title: Gluconeogenic enzyme PCK1 supports S-adenosylmethionine biosynthesis and promotes H3K9me3 modification to suppress hepatocellular carcinoma progression

doi: 10.1172/JCI161713

Figure Lengend Snippet: ( A ) Scheme of the experimental procedure with AlbCre –/– Pck1 fl/fl (WT), AlbCre +/– Pck1 fl/fl mice (liver-specific KO [LKO]) ( n = 6 mice per group). ( B ) Gross images and ( C ) quantification of weight and numbers of tumor nodules of livers from WT, Pck1 -LKO treated with vehicle, Pck1 -LKO treated with SAM, Pck1 -LKO injected with pSECC-sgControl, and Pck1 -LKO injected with pSECC-sg S100a11 mice ( n = 6 mice per group). ( D ) Representative images of H&E staining for lung tissues were provided, and ( E ) the metastatic foci were counted ( n = 6 mice per group). ( F ) Ultra-performance liquid chromatography results for the SAM concentration in mouse serum samples and liver tissues ( n = 6 mice per group). Scale bars: 200 μm ( D ). Data are shown as the mean ± SEM. Statistical analysis was performed using 1-way ANOVA with Tukey’s test ( C , E , and F ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: S100a11 sgRNA was cloned into the pSECC vector (a lentiviral vector encoding Cre and CRISPR components, Addgene plasmid 60820).

Techniques: Injection, Staining, Liquid Chromatography, Concentration Assay