Fig. 4: Intracisternal delivery of AAV-VEGFR3d1-4 impairs meningeal lymphatics and promotes stress-induced depression-like behaviors in male mice.
From: A functional role of meningeal lymphatics in sex difference of stress susceptibility in mice
a The experimental timeline of the AAV infusion, sub-chronic variable stress (SCVS), behavioral tests, trace injection into the cisterna magna (i.c.m.), and tissue collection (sac). b Representative images of eGFP (green)-labeled AAV9-infected cells surrounding LYVE1 (gray)-positive meningeal lymphatics (mLV). Nuclei: DAPI (blue). Left: whole mount dura mater, scale bars: 2000 µm. Right: enlarged views of the boxed areas from the image on the left, scale bars: 200 µm. c Representative images of the LYVE1 staining (gray) and the dextran tracer (red) in the SSS and COS + TS areas of dura mater, comparing male mice injected with AAV-VEGFR3d1-4 (VEGFR3d1-4 + SCVS) with those injected with AAV-VEGFR3d4-7 (VEGFR3d4-7 + SCVS). Both groups experienced SCVS. Scale bars: 500 µm. d Representative images of LYVE1+ mLV at higher magnification. Scale bars: 200 µm. e–h Quantification of the fluorescence intensity of the LYVE1 staining (e), the area covered by mLV (f), the diameter of LYVE1+ mLV (g), and the fluorescence intensity of the dextran tracer in the SSS and COS + TS areas of dura mater (h). i Representative images of dextran tracer (red) and DAPI (blue) in the dCLNs. Scale bars: 500 µm. j Quantification of the fluorescence intensity of dextran tracer in the dCLNs. k–n Quantification of grooming duration in the splash test (ST, k), the immobile time in the forced swim test (FST, l), the latency to eat in the novelty-suppressed feeding test (NSF, m) and changes in the body weight (n). o Representative traces of animal’s paths in the open field (OF). The center zone: the gray box in the center. p–r Quantification of the traveled distance in the center zone as a percentage of the total traveled distance (p), the time spent in the center zone as a percentage of the total time (q), and total traveled distance (r) in the OF (e–g, k–n, p–r: n = 8–9 per group; h, j: n = 7–8 per group; results from two independent experiments). All data are presented as mean ± s.e.m. and analyzed by unpaired Student’s t tests (e–h, j–l, n, p–r) or Mann–Whitney test (m). Source data are provided as a Source data file.
