sr57227a  (MedChemExpress)

Journal: International Journal of Molecular Sciences

Article Title: 5-HT3 Signaling Alters Development of Sacral Neural Crest Derivatives That Innervate the Lower Urinary Tract

doi: 10.3390/ijms22136838

Figure Lengend Snippet: Pharmacologically perturbing serotonin signaling alters the differentiation outcomes of sacral NCPs in vitro. ( A ) Sox10 -H2BVenus+ sacral NCPs were flow-sorted from 13.5 dpc pelvic ganglia and then grown in low-density cultures and treated with drug vehicle (negative control), SR57227A (specific 5-HT3 receptor agonist), or clozapine (mixed antagonist of several serotonin receptors). Mean ± SEM percentage of colonies expressing markers of differentiated cell types including neurons (Peripherin+, N), glia (GFAP+, G), and myofibroblast (Smooth Muscle Actin+, M) after exposure to either vehicle control, SR57227A, or clozapine are plotted. * p < 0.05, *** p < 0.001, assessed by Welch’s t -test. n = 33−45 wells per treatment condition. ( B ) Confocal images of 13.5 dpc Sox10 -H2B-Venus (green) pelvic ganglia explants cultured in the presence or absence of the 5-HT agonist SR57227A and subjected to detection of apoptotic cells (red, ApopTag) followed by immunohistochemistry with Hu C/D antibody, a pan-neuronal marker. Occasional apoptotic glial cells (arrowheads) and neuronal cells (arrows) are indicated. The scale bar for all insets is 20 μm. Representative DAPI staining reveals the explants also contain a non-glial, non-neuronal cell population and that the majority of apoptotic events take place in this population (inset with asterisk, 120× magnification). ( C ) Quantification of proportions of ApopTag+ cells out of total cells in pelvic ganglia explants. p = 0.99, Welch’s t -test.

Article Snippet: For 5-HT3 agonist-treated collagen gels, SR57227A (Sigma, S1688, St. Louis, MO, USA) was reconstituted in sterile MilliQ water to 1000 µM and added to the media to achieve concentrations ranging from 10 to 50 µM.

Techniques: In Vitro, Negative Control, Expressing, Cell Culture, Immunohistochemistry, Marker, Staining

Journal: International Journal of Molecular Sciences

Article Title: 5-HT3 Signaling Alters Development of Sacral Neural Crest Derivatives That Innervate the Lower Urinary Tract

doi: 10.3390/ijms22136838

Figure Lengend Snippet: Over-stimulating 5-HT3 in cultured fetal pelvic ganglia explants diminishes neurite arbor complexity. Sox10 -H2BVenus 13.5 dpc pelvic ganglia explants treated with 5-HT3 receptor agonist SR57227A exhibit a severe reduction in the elaboration and complexity of neuronal processes compared to vehicle control. ( A – C ) Fluorescent microscope images of pelvic ganglia explants treated with drug vehicle ( A ), 25 μm SR57227A ( B ), or 50 μm SR57227A ( C ). ( A’ – C’ ) Sholl masks generated for pelvic ganglia explant neuronal fiber tracings that demarcate concentric radii used to quantify neurite arbor complexity and growth. ( D ) Average number of intersections of the neurite arbor with Sholl radii plotted for each treatment group. ( E ) Average diameter of the outer-most circle drawn that encompasses the entire arbor. For both plots, black brackets are SEM. n = 12–14 pelvic ganglia explants per treatment group. *** = p < 0.001, n.s. = not statistically significant, assessed via one-way ANOVA with Tukey’s Honest Significant Difference test to correct for multiple comparisons.

Article Snippet: For 5-HT3 agonist-treated collagen gels, SR57227A (Sigma, S1688, St. Louis, MO, USA) was reconstituted in sterile MilliQ water to 1000 µM and added to the media to achieve concentrations ranging from 10 to 50 µM.

Techniques: Cell Culture, Microscopy, Generated