Journal: The Journal of Neuroscience

Article Title: A Distinct Metabolically Defined Central Nucleus Circuit Bidirectionally Controls Anxiety-Related Behaviors

doi: 10.1523/JNEUROSCI.1578-21.2022

Figure Lengend Snippet: sEH is enriched in neurons in the CeA and is associated with anxiety-related behaviors. A , left, Double immunofluorescence staining with sEH (red) and NeuN (green) reveals co-localization in the CeA of adult C57BL/6J mice. Scale bar: 100 μm. Right, Magnified view of left image. Scale bars: 50 μm (top) and 10 μm (bottom). B , left, Representative images of CeA Ephx2 (sEH)-tdTomato neurons (red) from the Ephx2-iCreER T2 ; Ai9 mice co-stained for NeuN (green). Scale bar: 100 μm. Right, Zoomed-out view of left image. Scale bars: 50 μm (top) and 20 μm (bottom). C , Representative images of CeA Ephx2 (sEH)-tdTomato neurons (red) from the Ephx2-iCreER T2 ; Ai9 mice co-stained for S100β (green). Scale bar: 100 μm. D , Percentage of sEH + neurons in the CeA that co-express NeuN and S100β ( t = 4.025, ** p = 0.0024, n = 6 slices from 3 mice). E , Single-molecule fluorescence in situ for sEH (green) and the indicated alternate cell markers-CRF (red), PKCδ (red), and SOM (red) in the CeA of C57 mice. Scale bars: 50 μm. F , Percentage of sEH + neurons in the CeA that co-express CRF, PKCδ, and SOM ( n = 3 sections per mouse, 3 mice per marker). G , Experimental design for identification of the high-anxiety and low-anxiety mice. H , Mice are divided into high-anxiety and low-anxiety subgroups in the EPM according to G . Time in the open arm, t ( 12) = 10.42, *** p < 0.0001; time in the closed arm, t (12) = 4.874, *** p = 0.0004. I , After the EPM test, the mice were screened again with the NSF according to G . t (12) = 6.25, *** p < 0.0001. J , Real-time quantitative fluorescence PCR analysis shows CeA sEH mRNA expression in high-/low-anxiety mice. t (12) = 3.052, * p = 0.0101 < 0.05. K , Correlations between the sEH mRNA level and the time in open arms (%) of low-anxiety and high-anxiety mice. Pearson correlation coefficient, r = 0.5665, * p = 0.0101 < 0.05. Data are presented as the mean ± SEM. Two-tailed unpaired Student's t test, NS ≥ 0.05; * p ≤ 0.05, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Ephx2-iCreER T2 mice were constructed in Shanghai Model Organisms Center (Shanghai, China).

Techniques: Double Immunofluorescence Staining, Staining, Fluorescence, In Situ, Marker, Expressing, Two Tailed Test

Journal: The Journal of Neuroscience

Article Title: A Distinct Metabolically Defined Central Nucleus Circuit Bidirectionally Controls Anxiety-Related Behaviors

doi: 10.1523/JNEUROSCI.1578-21.2022

Figure Lengend Snippet: Bidirectional manipulations of the activity of sEH CeA neurons in regulation of anxiety-related behaviors. A , Stereotaxic infusion of AAV-hSyn-DIO-hM3D(Gq)-mCherry or AAV-hSyn-DIO-hM4D(Gi)-mCherry virus into the CeA of Ephx2-iCreER T2 mice with intraperitoneal injection of CNO (2 mg kg −1 ) to regulate neural activity. B , Schematic of virus injection, drug administration and behavior tests. C , hM3Dq-mCherry expression in sEH-positive neurons of the CeA. Scale bar: 100 μm. D , Representative traces of whole-cell recording in an acute slice from CeA sEH-positive neurons expressing hM3Dq-mCherry after bath application of 1 μ m CNO. E , G , Chemogenetic activation of sEH-positive neurons during EPM and NSF. Time in the open arm, t (27) = 2.321, * p = 0.02821 < 0.05; time in the closed arm, t (27) = 3.564, ** p = 0.0014 < 0.05 ( n = 15 for saline, n = 14 for CNO; E ); t (23) = 3.233, ** p = 0.0037 < 0.01 ( F ); t (23) = 0.338, p = 0.7387 > 0.05 ( n = 14 for saline, n = 11for CNO; G ). H , Chemogenetic activation of sEH-positive neurons during OFT. t (23) = 0.6932, p = 0.4951 > 0.05, n = 14 for saline, n = 11for CNO. I , hM4Di-mCherry expression in sEH-positive neurons of the CeA. Scale bar: 100 µm. J , Representative traces of whole-cell recording in an acute slice from CeA sEH-positive neurons expressing hM4Di-mCherry after bath application of 1 μ m CNO. K–M , Chemogenetic inhibition of sEH-positive neurons during EPM and NSF. Time in the open arm, t (12) = 2.731, * p = 0.0182 < 0.05; time in the closed arm, t (12) = 1.671, p = 0.1206 ( K ); t (18) = 2.191, * p = 0.0419 < 0.01 ( L ); t (18) = 1.004, p = 0.3289 > 0.05 ( M ). N , Chemogenetic inhibition of sEH-positive neurons during OFT. t (24) = 1.321, p = 0.1989 > 0.05. Data are presented as the mean ± SEM. Two-tailed unpaired Student's t test, NS ≥ 0.05; * p ≤ 0.05, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: Ephx2-iCreER T2 mice were constructed in Shanghai Model Organisms Center (Shanghai, China).

Techniques: Activity Assay, Virus, Injection, Expressing, Activation Assay, Saline, Inhibition, Two Tailed Test