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Fig. 6 | Molecular Brain

Fig. 6

From: Proteins secreted by brain arteriolar smooth muscle cells are instructive for neural development

Fig. 6

Different organ-derived VSMC cell lines have diverse effects on neural development. a Representative images of hippocampal neurons treated with HAVSMC-CM, HBVSMC-CM, and HUVSMC-CM for 20 h starting at 4 h after plating. anti-Tuj1 (green), Hoechst (blue). b Cumulative probability of neurons with different neurite numbers. Insert: percentages of unipolar neurons in the whole population of neurons calculated from the same culture. c Representative images of hippocampal neurons with the same treatments as a with an additional 44 h after plating. d Cumulative probability in c. Insert: percentages of unipolar neurons in different group. e, f Quantification of average total neurite length per neuron in different group after treated for 20 h (e) and 44 h (f). g Raster plots generated from MEA measurements of primary hippocampal neurons at DIV14 with neurobasal A medium (control) or HBVSMC-, HAVSMC- and HUVSMC-derived conditioned media. Each black line represents a spike that crossed the noise threshold (± 6 × standard deviation). Blue lines represent a single-electrode burst—a collection of at least 5 spikes within an interspike interval (ISI) of 100 ms. Synchronized bursts are marked with purple boxes and defined as a collection of at least 10 spikes from a minimum of 25% of participating electrodes across the entire well, each separated by an ISI within 100 ms. Each plot represents one well from each control or treatment group during 300 s of recording. h, i Line charts showing parameters of neuronal functions, including the number of spikes (h) and weighted mean firing rate (i). MEA recordings were collected at DIV4, 7, 10, and 14 of primary hippocampal neuron cultures and analyzed using 10 min of recordings (Axion Biosystems). j NeuN+ cell densities of each group were compared at DIV21. One-way ANOVA with Dunnett’s post hoc test was used for statistical analysis. Mean ± SEM. k Bar charts exhibiting synchronized bursts at DIV14. Mean ± SEM. Three independent experiments were performed. Data are shown as the mean ± SEM. ***P < 0.001, ****P < 0.0001, ##P < 0.01

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