Fig. 1: Characterization of the CeL-specific Wfs1-FlpoER driver line. | Nature Communications

Fig. 1: Characterization of the CeL-specific Wfs1-FlpoER driver line.

From: Food and water intake are regulated by distinct central amygdala circuits revealed using intersectional genetics

Fig. 1: Characterization of the CeL-specific Wfs1-FlpoER driver line.

a Wfs1-FlpoER BAC transgene. Abbreviations: FlpoER optimized Flp-estrogen receptor fusion protein, NLS nuclear localization sequence, WPRE-pA woodchuck hepatitis post-transcriptional regulatory element with bovine growth hormone polyadenylation signal, gb2neo gb2 prokaryotic promoter driven Neo cassette. b Example showing endogenous Wfs1 immunoreactivity in the CeL. c Intersectional strategy used with Htr2a-Cre;Wfs1-FlpoER and Sst-Cre;Wfs1-FlpoER mice. Cells in CeL are either FlpoER positive or FlpoER and Cre double positive. Cells in CeM are only Cre-positive. d Examples of INTRSECT viruses. Con/Fon viruses express in CeL cells that are double positive for Cre and FlpoER, while Con/Foff viruses express in CeM cells that are Cre-positive and FlpoER-negative. e, t Expression through anterior-posterior axis [Bregma −0.9 (e, i, m, q) and −1.2 (f, j, n, r)] of Cre-dependent AAV-DIO-mCherry (not intersectional, red) in Htr2a-Cre;Wfs1-FlpoER mice showing the entire population of CeAHtr2a cells in the CeL and CeM (e, f), and expression of AAV-Con/Fon-EYFP (green) in the same mice predominantly highlighting the CeL fraction of Htr2a positive cells (e, f). Quantifications of the fraction of Htr2a-Cre::Con/Fon-EYFP positive cells in the CeL (54.3 ± 7.5%) among all CeAHtr2a cells in CeL (g) and distribution of Htr2a-Cre;Con/Fon-EYFP positive cells in CeL and CeM (h). i, j Expression of AAV-Con/Foff-EYFP (CeM enriched, green) versus AAV-DIO-mCherry (red) as described for (e, f). Quantifications of the fractions of Htr2a-Cre::Con/Foff-EYFP positive cells in CeM (66.3 ± 2.0%) among all CeAHtr2a cells in CeM (k) and distribution of Htr2a-Cre;Con/Foff-EYFP positive cells in CeL and CeM (l). m, n Expression of AAV-Con/Fon-EYFP (CeL enriched, green) versus AAV-DIO-mCherry (red) in Sst-Cre;Wfs1-FlpoER mice. o, p Quantification of the fraction of Sst-Cre::Con/Fon-EYFP positive cells in CeL (47.5 ± 7.7%) among all Sst-positive cells in CeL (o) and distribution of Sst-Cre;Con/Fon-EYFP positive cells in CeL and CeM (p). q, r Expression of AAV-Con/Foff-EYFP (CeM enriched, green) versus AAV-DIO-mCherry (red) in Sst-Cre;Wfs1-FlpoER mice. Quantification of the fraction of Sst-Cre::Con/Foff-EYFP positive cells in CeM (68.6 ± 5%) among all CeASst cells in CeM (s) and distribution of Sst-Cre;Con/Foff-EYFP positive cells in CeL and CeM (t) (all groups: n = 3 mice/ 9 sections). Values = Mean ± SEM. Scale bar: 115 μm.

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