Journal: Cell reports

Article Title: AAV5-mediated manipulation of insulin expression in choroid plexus has long-term metabolic and behavioral consequences

doi: 10.1016/j.celrep.2023.112903

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal Rabbbit anti-TTR (prealbumin) AbCam Cat # ab75815; RRID:AB_1310604 Monoclonal Rabbit anti-Insulin Cell Signaling Cat # 3014; RRID:AB_2126503 Polyclonal Rabbit anti-mCherry Rockland Cat # 600-401-P16; RRID:AB_2614470 Monoclonal Rabbit anti- p -Akt (Ser473) Cell Signaling Cat # 4060; RRID:AB_2315049 Monoclonal Mouse anti-Alpha1 Na+/K+ ATPase AbCam Cat # ab7671; RRID:AB_306023 Bacterial and virus strains AAV5-CMV-CRE Vector Biolabs Cat # 7012 AAV5-CMV-GFP Vector Biolabs Cat # 7006 AAV5-CMV-Ins2 Vector Biolabs Cat # AAV-262269 AAV5-CMV-GCaMP6f Vector Biolabs Built to order AAV5-TTR-hM3Dq-mCherry Vector Biolabs Built to order AAV5-CMV-Psck1 Vector Biolabs Cat # AAV-268239 AAV5-CMV-Pcsk2 Vector Biolabs Cat # AAV-268242 Chemicals, peptides, and recombinant proteins Compound 21 TOCRIS Cat # 5548 Critical commercial assays PicoPure RNA Isolation Kit ThermoFisher Cat # KIT0204 Ultra Sensitive Insulin ELISA Kit CrystalChem Cat # 90080 Deposited data Raw and analyzed data of RNA-seq This paper GEO: GSE218188 Experimental models: Cell lines iPS(IMR90)-4 WiCell WISCi004-B Experimental models: Organisms/strains C57BL/6J The Jackson Laboratory Strain # 000664 Ai14 The Jackson Laboratory Strain #007914 Ins1 −/− Ins2 fl/fl This paper N/A Software and algorithms ImageJ NIH https://imagej.nih.gov/ij/ GraphPad Prism GraphPad https://graphpad.com R R Foundation for Statistical Computing https://www.r-project.org Other Normal chow diet Dyets Inc Cat# 101845 High fat/high sugar diet Dyets Inc Cat# 103806 Open in a separate window KEY RESOURCES TABLE.

Techniques: Virus, Plasmid Preparation, Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Software

Journal: Cerebral Cortex (New York, NY)

Article Title: Activity of CaMKII-expressing neurons in medial prefrontal cortex of male and female Long-Evans rats is necessary for encoding odor information and novelty recognition in an odor-based incidental memory test

doi: 10.1093/cercor/bhaf203

Figure Lengend Snippet: Experimental design. (A). First, male and female rats underwent stereotaxis surgery. In experiment 1 the AAV9-CamKIIa-jGCaMP8m-WPRE vector was unilaterally infused into the prelimbic region of the mPFC [18 (9 per sex)]. In experiment 3, either a control (AAV5-CaMKII-GFP or AAV5-CaMKII-mCherry) or active vector (AAV5-CamKIIa-hM4D(Gi)-mCherry) was bilaterally infused into the prelimbic region of the mPFC [14 active (7 per sex), 10 control (5 per sex)]. Following a two-week recovery, the rats underwent behavioral testing. In experiment 1 calcium dynamics were recorded using fiber photometry and rats completed both the IST and DST with odors. In experiment 3, rats completed both the IST and DST with odors following either a saline or C21 i.p. injection (1.0 mg/kg) in a pseudorandom order. All brains were then sectioned using a cryostat or vibratome, and sections were stained with Hoescht. Lastly, a confocal microscope was used to image the brain sections to confirm the vector placement and expression. (B). An example of a brain from experiment 1 with adequate vector expression and location. (C). An example of a brain from experiment 3 with adequate vector expression and location. Images were taken using a confocal microscope at 5X using a 594 nm laser, and the prelimbic region of the mPFC (PrL) is labeled. Figure created with biorender.

Article Snippet: The inhibitory DREADD agonist C21 dihydrochloride (water soluble) (1 mg/kg, HelloBio HB6124), was delivered via an i.p. injection 1 h prior to behavioral testing.

Techniques: Plasmid Preparation, Control, Saline, Injection, Staining, Microscopy, Expressing, Labeling

Journal: Cerebral Cortex (New York, NY)

Article Title: Activity of CaMKII-expressing neurons in medial prefrontal cortex of male and female Long-Evans rats is necessary for encoding odor information and novelty recognition in an odor-based incidental memory test

doi: 10.1093/cercor/bhaf203

Figure Lengend Snippet: Compound 21 (C21) does not significantly affect novel odor preference in the IST or DST with odors. Naïve adult male (A) and female (B) rats was tested. There was no observed difference in novelty preference between saline or C21 treatment conditions in either sex, or when samples were combined (C).

Article Snippet: The inhibitory DREADD agonist C21 dihydrochloride (water soluble) (1 mg/kg, HelloBio HB6124), was delivered via an i.p. injection 1 h prior to behavioral testing.

Techniques: Saline

Journal: Cerebral Cortex (New York, NY)

Article Title: Activity of CaMKII-expressing neurons in medial prefrontal cortex of male and female Long-Evans rats is necessary for encoding odor information and novelty recognition in an odor-based incidental memory test

doi: 10.1093/cercor/bhaf203

Figure Lengend Snippet: Role of the mPFC in the IST and DST with odors. Rats performed the IST and DST with odors following both a saline and C21 i.p. injection (1 mg/kg). A discrimination ratio (DR) was generated to evaluate novelty preference (DR closer to 1). In the IST, there was no difference in novelty preference following the two treatments in rats with either the control vector (A) or the active vector (B) expressed in the mPFC. In the DST with odors, there was no difference in novelty preference following the two treatments in rats with the control vector (A) expressed in the mPFC. However, there was a significant decrease following an i.p. injection of C21 (1.0 mg/kg) in the rats with the active vector (B) expressed in the mPFC. This indicates a role for the mPFC in supporting odor-based incidental memory under higher memory loads. ** P < 0.01.

Article Snippet: The inhibitory DREADD agonist C21 dihydrochloride (water soluble) (1 mg/kg, HelloBio HB6124), was delivered via an i.p. injection 1 h prior to behavioral testing.

Techniques: Saline, Injection, Generated, Control, Plasmid Preparation

Journal: Cerebral Cortex (New York, NY)

Article Title: Activity of CaMKII-expressing neurons in medial prefrontal cortex of male and female Long-Evans rats is necessary for encoding odor information and novelty recognition in an odor-based incidental memory test

doi: 10.1093/cercor/bhaf203

Figure Lengend Snippet: Examining c-Fos as a biomarker for neural activity in the mPFC. Rats performed the DST with odors following either a saline and C21 i.p. injection (1 mg/kg) to activate the hM4D(Gi) DREADD expressed in the mPFC. (A) There was a decrease in the number of c-Fos positive cells in the mPFC- based ROI following an i.p. injection of C21. (A) There was also a significant increase in the number of c-Fos positive cells in comparison to home cage rats following exposure to the DST test when using the mPFC-based ROI for analysis. An i.p. injection of C21 prevented this increase. (B) A visual decrease, albeit not significant, is observed following an i.p. injection of C21 in the number of c-Fos positive cells in the vector cloud- based ROI. (C, D) As well, a visual decrease is observed following an i.p. injection of C21 in the number of colocalized cells. The prelimbic region of the mPFC (PrL) is labeled. * P < 0.05.

Article Snippet: The inhibitory DREADD agonist C21 dihydrochloride (water soluble) (1 mg/kg, HelloBio HB6124), was delivered via an i.p. injection 1 h prior to behavioral testing.

Techniques: Biomarker Discovery, Activity Assay, Saline, Injection, Comparison, Plasmid Preparation, Labeling

Journal: Biological Psychiatry Global Open Science

Article Title: Activity in the Dorsomedial Striatum Underlies Serial Reversal Learning Performance Under Probabilistic Uncertainty

doi: 10.1016/j.bpsgos.2022.08.005

Figure Lengend Snippet: Timeline of experimental manipulations. Top panel: Experiment 1. Mice were trained until PRL performance was stable and then underwent tract-tracing surgery. After recovery and rebaselining, mice were tested for each contingency (80:20, 70:30, 80:40, and 90:10), separated by 2 days. Following 5 additional days of baseline PRL testing, mice were administered amphetamine (1 mg/kg, intraperitoneally) or saline (15 minutes prior to testing) at an 80:20 contingency. Mice were perfused 60 minutes after completion of testing (i.e., 90 minutes after the middle of the reversal learning session) for subsequent cFos analysis. Bottom panel: Experiment 2. After DREADDs surgery and PRL training, mice underwent acute CNO administration (30 minutes prior to testing). This was conducted using a within-subject Latin-square design at the 80:20 contingency with CNO (0, 0.5, 1, and 2 mg/kg, in 0.5% DMSO and 0.9% saline). This was followed by a crossover design with CNO (0 and 2 mg/kg) at the 80:40 contingency. Subsequently, mice were given fresh water daily (0.25% DMSO) for 5 days at 80:20 followed by a water solution containing CNO (∼8 mg/kg/day). CNO exposure was continued for 23 days as follows: 14 days at 80:20, 4 days at 80:40, and 5 days at 70:30. After testing was complete, a small cohort of mice were administered 2 mg/kg CNO intraperitoneally and perfused 2 hours later for cFos analysis. CNO, clozapine N -oxide; DMSO, dimethyl sulfoxide; DREADD, designer receptors exclusively activated by designer drug; PRL, probabilistic reversal learning.

Article Snippet: DREADDs can be activated chronically via CNO (clozapine N -oxide) (AK Scientific) in drinking water , allowing us to manipulate these circuits acutely and chronically during reversal learning.

Techniques: Saline

Journal: Biological Psychiatry Global Open Science

Article Title: Activity in the Dorsomedial Striatum Underlies Serial Reversal Learning Performance Under Probabilistic Uncertainty

doi: 10.1016/j.bpsgos.2022.08.005

Figure Lengend Snippet: Effects of acute stimulation of midbrain projections to the DMS on reversal learning performance. Pathway-specific DREADDs were used to express hM3Dq receptors (or mCherry alone for control mice) in midbrain cells projecting to the DMS (A) . Administration of CNO increased cFos+ cell number (B) and cFos intensity (C) in mCherry-labeled cells in the midbrain of hM3Dq-expressing mice ( n = 2–3). Acute within-subject treatment with CNO (control mice n = 12, hM3Dq n = 6) did not alter the number of completed trials (D) , reversal performance (E) , or win-stay probability (F) at 80:20 or 80:40 contingencies. Activating cells projecting from the midbrain to the DMS significantly decreased lose-shift probability (G) at the 80:40 contingency. Data are expressed as mean ± standard error. ∗ p < .05, ∗∗ p < .01. CNO, clozapine N -oxide; DMS, dorsomedial striatum; DREADDs, designer receptors exclusively activated by designer drugs; TH, tyrosine hydroxylase.

Article Snippet: DREADDs can be activated chronically via CNO (clozapine N -oxide) (AK Scientific) in drinking water , allowing us to manipulate these circuits acutely and chronically during reversal learning.

Techniques: Control, Labeling, Expressing

Journal: Biological Psychiatry Global Open Science

Article Title: Activity in the Dorsomedial Striatum Underlies Serial Reversal Learning Performance Under Probabilistic Uncertainty

doi: 10.1016/j.bpsgos.2022.08.005

Figure Lengend Snippet: Effects of chronic stimulation of midbrain projections to the DMS on reversal learning performance. Experimental timeline featuring 5 days of baseline testing at 80:20 with new water bottles containing DMSO for acclimatization (A) . Mice were then given water bottles containing CNO and assessed for an additional 14 days at 80:20, followed by 4 days at 80:40 and 5 days at 70:30. All mice averages are represented as a percentage of their group’s baseline (e.g., controls normalized to control baseline [ n = 11] and hM3Dq normalized to hM3Dq baseline [ n = 6]). Chronic CNO treatment to stimulate midbrain to DMS pathways did not alter the average number of trials completed at any contingency (B) . DREADDs activation during both the 80:40 and 70:30 contingencies significantly decreased reversal performance (C) . No differences were observed for win-stay probability (D) , but DREADDs activation significantly decreased lose-shift probability during both the 80:20 and 80:40 contingencies (E) . Data are expressed as mean ± standard error. ∗ p < .05, ∗∗ p < .01, # p < .05 compared with hM3Dq baseline. CNO, clozapine N -oxide; DMS, dorsomedial striatum; DREADDs, designer receptors exclusively activated by designer drugs; PRL, probabilistic reversal learning.

Article Snippet: DREADDs can be activated chronically via CNO (clozapine N -oxide) (AK Scientific) in drinking water , allowing us to manipulate these circuits acutely and chronically during reversal learning.

Techniques: Control, Activation Assay