Biased signaling at NTSR1 differentially regulates inhibitory synaptic transmission in the extended amygdala and suppresses motivated feeding in mice

Figures

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  Figure 1.. SBI-553 blocks NTS-induced increases in spontaneous GABAergic neurotransmission onto CeAL neurons independently of sex (A) Schematic of CeA subregions across the AP axis and recording location of electrophysiology experiments denoting all recordings were in the CeAL in this figure (teal) (B) NTS increases the sIPSC frequency in both males and females. This effect returns to baseline levels during the washout. NTS did not change the (C) IPSC amplitude or (D) IPSC area in either sex (E) SBI-553 alone does not alter the IPSC frequency, but blocks NTS from increasing IPSC frequency. SBI-553 alone or in combination with NTS does not alter (F) IPSC amplitude or (G) IPSC area. Representative traces of (H) NTS perfusion and (I) SBI-553 and NTS perfusion. Error bars represent average value ± SEM. ** p<0.01

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  Figure 2.. NTS does not alter spontaneous GABAergic neurotransmission onto CeAM neurons (A) Schematic of CeA subregions across the AP axis and recording location of electrophysiology experiments denoting all recordings were in the CeAM (light blue) in this figure (B) Females have a higher IPSC frequency at baseline than males, but NTS does not alter the sIPSC frequency in either sex. NTS also does not alter the (C) IPSC amplitude or (D) IPSC area. Representative traces of NTS perfusion in (E) females and males. Error bars represent average value ± SEM. * p<0.05

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  Figure 3.. SBI-553 does not block NTS-induced increases in spontaneous GABAergic neurotransmission onto ovBNST neurons (A) Schematic of the ovBNST subnuclei and recording location of electrophysiology experiments (teal) (B) Representative traces of SBI-553 and NTS perfusion (C) NTS increases the sIPSC frequency in both males and females. This effect returns to baseline levels during the washout. NTS differentially alters the (D) IPSC amplitude or (E) IPSC area in males and females (F) SBI-553 alone does not alter the IPSC frequency. SBI-553 is unable to block NTS-induced increases in IPSC frequency, but blocks NTS-induced increases in (G) IPSC amplitude and (H) IPSC area in the males. Representative traces of (I) NTS perfusion in males and females. Error bars represent average value ± SEM. ** p<0.01

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  Figure 4.. Single injection of SBI-553 alters the cFos immunofluorescence in the CeA, but not the BNST of naïve animals. (A) SBI-553 does not alter cFos expression in the CeAC (B) or CeAM of naïve animals (C) but selectively increases activity in the CeAL of female mice compared to vehicle treatment. (D) Representative immunofluorescence images of cFos expression (red) and DAPI (blue) in the CeA. (E) SBI-553 has no impact on cFos activity in the dBNST, (F) vBNST, (G) or ovBNST of naïve male or female mice. (H) Representative immunofluorescence images of cFos expression in the BNST. Error bars represent average value ± SEM. * p<0.05

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  Figure 5.. SBI-553 decreases palatable food consumption in both male and female mice in the Motivated Homecage Feeding Assay (MHFA). (A) Schematic of the MHFA timeline. (B) SBI-553 reduces palatable motivated feeding in both male and female mice. Error bars represent average value ± SEM. ** p<0.01, **** p<0.0001

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  Figure 6.. SBI-553 alters the cFos immunoflourescence in the CeA, but not the BNST after the MHFA. (A) SBI-553 significantly increases cFos expression in the CeAc, especially in females, (B) but not the CeM (C) or CeAL of any mice after the MHFA compared to vehicle treatment. (D) Representative immunofluorescence images of cFos expression (red) and DAPI (blue) in the CeA. (E) SBI-553 has no impact on cFos activity in the dBNST, (F) vBNST, (G) or ovBNST of male or female mice after the MHFA. (H) Representative immunofluorescence images of cFos expression in the BNST. Error bars represent average value ± SEM. * p<0.05

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  Figure 7.. SBI-553 reduces palatable FL consumption in both fed and fasted conditions. (A) Schematic of the FvFHFA timeline. (B) SBI-553 significantly reduces feeding behavior in both fed and fasted feeding states, with (C) a larger reduction in in the fasted animals. (D) There was no difference in treatment order on reductions of consumption across feeding conditions. Error bars represent average value ± SEM. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001

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  Figure 8.. SBI-553 alters the cFos immunoflourescence in the CeA and the ovBNST after the FvFHFA. (A) SBI-553 significantly increases cFos expression in the CeAC and (C) CeAL, (B) but not the CeAM compared to vehicle treatment across feeding conditions. (D) Representative immunofluorescence images of cFos expression (red) and DAPI (blue) in the CeA. (E) SBI-553 has no impact on cFos activity in the dBNST in either fed or fasted animals in the FvFHFA. (F) In the vBNST, fasted animals showed higher levels of cFos activity than fed animals, which was significantly different between animals receiving vehicle or SBI-553. (G) ovBNST cFos activity not impacted by SBI-553 treatment in fasted animals but showed trending elevations in animals receiving SBI-553 in a fed state compared to vehicle, with a significant interaction between SBI-553 and feeding condition (two-way ANOVA, interaction p=0.04). (H) Representative immunofluorescence images of cFos expression in the BNST. Error bars represent average value ± SEM. *