dynorphin pro dynorphin  (Neuromics)

Journal: Nature Communications

Article Title: The claustrum-prelimbic cortex circuit through dynorphin/κ-opioid receptor signaling underlies depression-like behaviors associated with social stress etiology

doi: 10.1038/s41467-023-43636-x

Figure Lengend Snippet: a Representative immunoblots of dynorphin expression in the PL. b Quantification of dynorphin A ( n = 9–10, t (17) = 2.655, P = 0.0167. Student’s t test). c Representative immunoblots of KOR expression in the PL. d Quantification of KOR ( n = 6, t (10) = 0.6318, P = 0.5417. Student’s t test). e , l Schematic of CLA injection of AAV2/9-CaMKIIα-DIO-Oprk1-P2A-mCherry and AAV2/9-CaMKIIα-ChR2-EYFP and PL injection of AAV2/R-CaMKIIα-Cre-WPRE-hGH in WT mice ( e ) or CLA injection of AAV2/9-hSyn-fDIO-CRE-mCherry-WPRE-hGH and AAV2/9-CaMKIIα-ArchT-EGFP and PL injection of AAV2/R-CaMKIIα-FLP-WPRE-hGH in Oprk1 lox/lox mice ( l ). f , m In situ hybridization of KOR (white), vGlut1 (green), mCherry (red) and DAPI (blue) in the CLA. g , n Quantification of KOR colocalization with vGlut1 and mCherry ( g n = 3, t (4) = 4.288, P = 0.0128; n n = 3, t (4) = 4.573, P = 0.010. Student’s t test). h – k KOR over-expression in CLA Glu -PL circuit blocked photo-activation of the CLA Glu -PL pathway induced increase of social interaction ratio ( h n = 9–13, F (1,40) = 7.733, P = 0.0082. Two-way ANOVA), increase of sucrose consumption in the SPT ( i n = 10–12, F (1,40) = 8.265, P = 0.0064. Two-way ANOVA) and decrease of immobility time in the TST ( j n = 11–14, F (1,46) = 5.662, P = 0.0215. Two-way ANOVA) without affecting locomotor activity ( k n = 13–14, t (25) = 1.837, P = 0.0781. Student’s t test). o – r knockdown of KOR in CLA Glu -PL circuit reversed photo-inhibition of CLA Glu -PL induced decrease of social interaction ratio ( o n = 17–18, F (1,66) = 0.2481, P = 0.6201. Two-way ANOVA), decrease of sucrose consumption in the SPT ( p n = 15–16, F (1,59) = 5.132, P = 0.0272. Two-way ANOVA) and increase of immobility time in the TST ( q n = 17–19, F (1,68) = 5.837, P = 0.0184. Two-way ANOVA) without affecting locomotor activity ( r n = 15–17, t (30) = 0.06621, P = 0.9477. Student’s t test). All data are shown as mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: A 30 μg protein aliquot from each sample was separated using SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane, which was then blocked with 5% BSA in 0.1% Tween 20 in TBS (TBST) for 2 h. The membranes were then incubated with primary antibodies against dynorphin A (1:300; Abcam, ab82509) and β-actin (1:3000, Cell Signaling Technology, 3700) overnight at 4 °C.

Techniques: Western Blot, Expressing, Injection, In Situ Hybridization, Over Expression, Activation Assay, Activity Assay, Inhibition

Journal: Nature Communications

Article Title: The claustrum-prelimbic cortex circuit through dynorphin/κ-opioid receptor signaling underlies depression-like behaviors associated with social stress etiology

doi: 10.1038/s41467-023-43636-x

Figure Lengend Snippet: Top panel, PL-projecting CLA glutamatergic neurons express KORs, synapse onto both excitatory pyramidal neurons (PNs) and PV interneurons in the PL and play a key role in the maintenance of the E/I balance in the PL through driving feedforward inhibition (FFI) of PNs by excitation of PV interneurons. Presynaptic KOR signaling profoundly inhibits the excitatory transmission from the CLA to PL PV neurons (Supplementary Fig. ). Bottom panel, Chronic stress results in endogenous dynorphin release and increase KOR signaling in the PL. More strongly increasing KOR signaling of CLA synapses in PL PV interneurons lead to preferential impairment of excitatory transmission from the CLA to PL PV interneurons, which in turn induces the hyper-excitation of PL PNs as a result of the loss of FFI of PV interneurons on PNs in the PL, thereby resulting in E/I imbalance in the PL and depression. Source data are provided as a Source Data file.

Article Snippet: A 30 μg protein aliquot from each sample was separated using SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane, which was then blocked with 5% BSA in 0.1% Tween 20 in TBS (TBST) for 2 h. The membranes were then incubated with primary antibodies against dynorphin A (1:300; Abcam, ab82509) and β-actin (1:3000, Cell Signaling Technology, 3700) overnight at 4 °C.

Techniques: Inhibition, Transmission Assay

Journal: Cell reports

Article Title: Dissecting the Roles of GABA and Neuropeptides from Rat Central Amygdala CRF Neurons in Anxiety and Fear Learning

doi: 10.1016/j.celrep.2019.08.083

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Guinea pig polyclonal anti-Dynorphin , Neuromics , Cat# GP10110; RRID:AB_1621439.

Techniques: Virus, Recombinant, RNAscope, Multiplex Assay, cDNA Synthesis, Gene Expression, Bicinchoninic Acid Protein Assay, Software