Journal: bioRxiv
Article Title: Choroid Plexus Hosts the Earliest Detectable Circadian Clock in the Brain
doi: 10.1101/2025.09.09.675224
Figure Lengend Snippet: ( A ) Schematic of the protocol used for recording PER2::LUC bioluminescence from embryonic brain or head slices (left) or from the isolated 4VCP (right). ( B ) Illustration summarizing the finding that the 4VCP is circadian earlier than the SCN in the mouse embryo. ( C ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E15.5 brain slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : Images of immunofluorescence of LHX1 (left) and TTR (right) after brain clearing d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( D ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E13.5 head slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : PER2::LUC images with white boxes indicating the putative hypothalamus (left) and 4VCP (right). d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( E ) a : Bright field and PER2::LUC images of the bioluminescence obtained from a PER2::LUC E11.5 head slice. b : Analysis of the recording showing the period (left) and the circadian power (right). c : PER2::LUC images with white boxes indicating the putative hypothalamus (left) and 4VCP (right). d : Detrended time series of PER2::LUC bioluminescence from the areas indicated by the white boxes in c. ( F ) a : PER2::LUC image of the bioluminescence obtained from a PER2::LUC E13.5 isolated 4VCP. b : Analysis of the recording showing the period (left) and the circadian power (right). c : Detrended time series of the recording. Scale bar: 500 µm. Red arrows indicate 4VCP and white arrows indicate SCN.
Article Snippet: Similar steps of wash and blocking were applied before consecutive incubations with LHX1 (AB_531784; 4F2-c, mouse, 1:500, DSHB) and anti-mouse CF350 (SAB4600222, goat, 1:1000, Sigma) as before.
Techniques: Isolation, Slice Preparation, Immunofluorescence