Figure 8.
Peptidergic neurons of the EWcp enable preparatory nesting through the modulation of a broad network of downstream arousal and nesting centers in the brain
(A) Whole brain mapping of labeled EWcpCART+ projections with iDISCO+ clearing. Sagittal projection of the light sheet data is shown.
(B) Atlas-aligned raw data (coronal projections).
(C) Segmented projections of EWcpCART+ neurons using TrailMap and ClearMap, showing dense projections in the brainstem and hypothalamus.
(D) Coronal section of the segmented data highlighting regions of dense projections.
(E) Strategy to map the brain-wide activation pattern elicited by the stimulation of EWcpCART+ neurons in hM3Dq- or mCherry-injected pregnant mice.
(F) Sections at the level of the EW in mice injected with CNO or saline, immunolabeled for Fos and mCherry, showing the induction of Fos expression following CNO injection.
(G) Whole-brain mapping with iDISCO+ of Fos expression evoked by CNO injections in E15 pregnant females over saline-injected pregnant females, showing induction of Fos in the EW.
(H) Combined whole brain datasets of projection and activation patterns.
(I) Overlap of projection and activation maps reveals regions downstream of the EWcpCART+ neurons in the preoptic areas and brainstem reticular formation.
(J) Volcano plot of Fos expression in brain regions induced by CNO over saline in CART-cre mice injected with AAV2-DIO-HM3Dq. Regions highlighted in green are also significantly activated during nesting (based on voxel maps) and receive projections from EWcpCART+ neurons. Scale bars are 1 mm, except (F): 50 μm. Student's t test for (F), (I), and (J). See also Video S4.