Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: a . Representative light-sheet microscopy images of the somatosensory cortical endothelial network immunolabeled for CD31 (in grey) on P5 (whole cleared brain) and spinning-disk microscopy images of somatosensory cortical sections on P15 and P30 in males and females. Scale bars: 100 µm for P5 and 50 µm on P15 and P30. b. A 3D reconstruction of the filaments corresponding to vessels. Scale bars: 50 µm. c-g. Quantification of the vascular network in the somatosensory cortex. Length (c), number of segments (d); segment length ( e ); branch angle ( f ); segment straightness ( g ). n P5 male = 4, n P5 female = 3, n P15 male = 4, n P15 female = 4, n P30 male = 4, n P30 female = 4. The data are presented as the mean ± SD and were analyzed using an unpaired t-test . h. Representative spinning-disk images of pericytes immunolabeled for CD13 (in magenta) in somatosensory cortex sections on P15. Blood vessels are stained with isolectin-B4 (IB4, in green) in males and females. Scale bars: 50 µm. i . Enlarged view of a pericyte. Scale bar: 10µm j. Quantification of CD13 perivascular coverage. n= 4 for each sex and stage. Dark dots represent the mean value for each mouse. Light dots represent values for each analyzed photos (4 images per mouse). The data are presented as the mean ± SD and were analyzed using a chi-squared test. The raw data are given in Source Data Table S5.

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: Microscopy, Immunolabeling, Staining

Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: Representative spinning-disk images of pericytes immunolabeled for CD13 (in magenta) in somatosensory cortex sections on P5, P15, P30 and P60 in males and in females. Blood vessels are stained with isolectin—B4 (IB4, in green) from P5 to P30 and with CD31 (in green) on P60. Scale bar: 50 µm.

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: Immunolabeling, Staining

Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: a . Representative spinning-disk images of astrocyte perivascular processes immunolabelled for Aqp4 (in green) in male and female somatosensory cortex sections on P15. Vessels are stained with IB4 (in grey) Scale bars: 50 µm. b. Quantification of perivascular Aqp4 on P5, P15, and P30. n= 4 for each sex and stage. Dark dots represent the mean value for each mouse and light dots represent values for each analyzed image (4 images per mouse). The data are presented as the mean ± SD and were analyzed using a chi-squared test. c. left , representative image of Sox9 immunolabelling in the somatosensory cortex (in grey) on P5; right, Cellpose detection of Sox9+ nuclei. Scale bars: 50 µm. d. Representative images of Sox9+ cells (in magenta) in male and female on P5, P15, and P30. Scale bar: 50 µm. e. Males and females did not differ in the Sox9+ cell density in the somatosensory cortex during development. n P5 male = 3, n P5 female = 3, n P15 male = 4, n P15 female = 3, n P30 male = 4, n P30 female = 3, n P60 male = 4, n P60 female = 4. The data are presented as the mean ± SD and were analyzed using a Mann-Whitney test. The raw data are given in Source Data Table S5.

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: Staining, MANN-WHITNEY

Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: Representative spinning-disk images of astrocyte perivascular processes immunolabeled for AQP4 (in green) in somatosensory cortex sections on P5, P15, P30 and P60 in males and in females. Blood vessels are stained with isolectin—B4 (IB4, in grey) from P5 to P30 and with CD31 (in grey) on P60. Scale bar: 50 µm. c. Representative spinning-disk images of Sox9+ cells (in magenta) in somatosensory cortex sections on P5, P15, P30 and P60 in males and in females. Scale bar: 50 µm.

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: Immunolabeling, Staining

Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: a-b . Representative images showing fluorescent in situ hybridization detection of Col1a1 + PVFs (orange) on P15 in somatosensory cortex sections from males and females. Vessels were stained with CD31 (in grey). Scale bar: 200 µm (left panel) and 100 µm (right panel) c . Quantification of Col1a1 + PVF density on P15. n male = 4, n female = 4. The data are presented as the mean ± SD and were analyzed using a Mann-Whitney test. d-e . Representative images of PVMs immunolabeled for CD206 (in red) and Lyve-1 (in cyan) on P15 in somatosensory cortex sections from males and females. Scale bar: 500 µm with close-ups on some blood vessels (I-II). Scale bar: 50 µm. f . CD206 + PVM density n male = 4, n female = 4. g . Lyve-1 + PVM density. n male = 8, n female = 8. h . The percentage of blood vessels with at least one PVM in the somatosensory cortex. n male = 8, n female = 8. The data are presented as the mean ± SD and were analyzed using an unpaired t-test. i . Zoom on a blood vessel with co-immunolobelled CD206 + Lyve-1 + PVM. Scale bar: 50µm. j . Zoom on a blood vessel with a CD206 + Lyve-1 - PVM. Scale bar: 50 µm. k . Zoom on a blood vessel with a CD206 - Lyve-1 + PVM. Scale bar : 50 µm. m . Counts of CD206 + Lyve-1 + , CD206 + Lyve-1 - and CD206 - Lyve-1 + PVM. n male = 4, n female = 4. The data are presented as the mean ± SD and were analyzed using a Mann-Whitney test. The raw data are given in Source Data Table S5

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: In Situ Hybridization, Staining, MANN-WHITNEY, Immunolabeling

Journal: bioRxiv

Article Title: Sex influences gliovascular unit assembly and function in the developing mouse brain

doi: 10.64898/2026.04.13.718096

Figure Lengend Snippet: a. Representative light-sheet microscopy images of whole cleared somatosensory cortex immunolabeled for smooth muscle actin (SMA; in cyan) on P5, P15, and P30 in males and females ( left panel ) and their respective 3D reconstructions ( right panel ). b–e Quantification of SMA⁺ blood vessels: total vessel length ( b ); total number of vessels ( c ); number of secondary branches ( d ); number of SMA + vessels with at least one secondary branch ( e ). The data are presented as the mean ± SD and were analyzed using an unpaired t-test. n P5 male = 4, n P5 female = 4, n P15 male = 3, n P15 female = 4, n P30 male = 4, n P30 female = 4. f. Representative map of in vivo arterial spin labelling MRI acquisition in males and females on P15 g . Analysis of CBF in the dorsal and lateral cortex. Dark dots represent the mean value for each mouse, and light dots represent values for each hemisphere. The data are presented as the mean ± SD and were analyzed using a one-tailed, unpaired t-test. n male = 4, n female = 4. h . Immunohistochemical analysis of SMA expression in the developing cortex in human males and females. i . Quantification of SMA-stained surface areas, quoted as the mean ± SD when possible. The low number of samples prevented the application of statistical tests. The raw data are given in Source Data Table S5 .

Article Snippet: 3D reconstructions of the somatosensory cortex were produced using Imaris software (Oxford Instruments, Oxford, UK).

Techniques: Microscopy, Immunolabeling, In Vivo, One-tailed Test, Immunohistochemical staining, Expressing, Staining