acsf solution  (Tocris)

Journal: Nature Communications

Article Title: A cell-type-specific circuit of somatostatin neurons in the habenula encodes antidepressant action in male mice

doi: 10.1038/s41467-025-58591-y

Figure Lengend Snippet: a – c Schematic diagram ( a ), representative images showing the expression of SST GFP neurons ( b ) and the axon projection targets of HBX SST neurons ( c ) ( n = 3 mice). d – f Schematic diagram ( d ), representative images ( e , n = 3 mice), and a plot ( f ) showing the axon projection targets of HBX SST neurons and c-fos anti-body staining is utilized to identify the activation of neurons within the IPN. Data are presented as mean ± SEM ( n = 3 mice per group, ** p = 3.78 × 10 −4 , two-tailed t-test). g – k SST neurons project excitatory synapses onto PENK neurons. Schematic diagram ( g ) and representative images showing the expression of ChR2-GFP in SST neuronal axon terminals ( h , n = 3 mice) and tdT in PENK neurons. Whole-cell patch-clamp recordings from PENK neurons were validated by biocytin labeling through recording electrodes ( i , n = 4 mice). The evoked EPSCs by optogenetic stimulation of axon terminals of SST ChR2GFP neurons were completely blocked by 20 μM CNQX ( j ) or 1 μM TTX and reversed by 4-AP ( k ). Data are presented as mean ± SEM ( n = 10 neurons from 7 mice, *** p = 3.96 × 10 −11 in ( j ), ACSF vs. TTX *** p = 2.68 × 10 -9 in ( k ), TTX vs. TTX/4-AP *** p = 1.75 × 10 -6 in ( k ), paired sample two-tailed t-test for ( j ), one-way repeated measures ANOVA for ( k ), ns, no significant difference). l – o SST neurons receive inhibitory synaptic inputs from CCK neurons. Schematic diagram ( l ) and representative images showing the expression of TVA/G and GFP in SST neurons ( m , n = 3 mice) and GFP in CCK neurons ( n , n = 3 mice). The evoked IPSCs ( o ) recorded from SST neurons were completely blocked by 20 μM BIC or 1 μM TTX (also see Supplementary Fig. ). Data are presented as mean ± SEM ( n = 10 neurons from 6 mice, ACSF vs. TTX *** p = 5.07 × 10 −12 , ACSF vs. BIC *** p = 5.01 × 10 −10 , paired sample two-tailed t-test). VTA: ventral tegmental area; IPN: interpeduncular nucleus; PENK: proenkephalin; ACSF: artificial cerebrospinal fluid; CNQX: 6-cyano-7-nitroquinoxaline-2,3-dione; TTX: tetrodotoxin; 4-AP: 4-amino phenylacetic acid; BNST: bed nucleus of the stria terminalis; BIC: bicuculline. Source data are provided as a Source Data file.

Article Snippet: For recording SST→PENK synaptic transmission, the external ACSF solution contained GABA A -receptor antagonists including 20 μM bicuculline (TOCRIS, 0130).

Techniques: Expressing, Staining, Activation Assay, Two Tailed Test, Patch Clamp, Labeling