Journal: Journal of neurobiology

Article Title: Hedgehog and Fgf Signaling Pathways Regulate the Development of tphR -Expressing Serotonergic Raphe Neurons in Zebrafish Embryos

doi: 10.1002/neu.20023

Figure Lengend Snippet: Inhibition of Fgf receptor activity leads to loss of serotonergic raphe neurons. Dorsal views of tphR expression and 5HT-IR in the brains of wild-type embryos treated with 9 μM SU5402 from 24 hpf to the stage indicated bottom left. Arrows point to serotonergic raphe neurons.

Article Snippet: SU5402 and Cyclopamine Treatment Dechorionated 24 h postfertilization (hpf) embryos were incubated in the dark in 9 μM SU5402 (Calbiochem) or 100 μM cyclopamine (Toronto Research Chemicals) until 48 hpf for in situ hybridization and 60 hpf for 5HT antibody staining.

Techniques: Inhibition, Activity Assay, Expressing

Journal: Development (Cambridge, England)

Article Title: The mevalonate pathway is a crucial regulator of tendon cell specification

doi: 10.1242/dev.185389

Figure Lengend Snippet: A zebrafish chemical screen identifies statins as regulators of craniofacial and pectoral fin tendon progenitors. (A) Design of the small-molecule screen. Wild-type embryos were incubated with individual compounds from 32 to 56 hpf and alterations in craniofacial scxa expression were assessed at 56 hpf. (B-E) Craniofacial scxa expression at 56 hpf, after incubation from 32 to 56 hpf (B,C) and at 72 hpf after incubation from 48 to 72 hpf (D,E). Atorvastatin expanded scxa expression in the pharyngeal arches (arrows) compared with controls. (F-I) Pectoral fin scxa:mcherry expression at 56 hpf (F,G) and 72 hpf (H,I) upon incubation from 32 to 56 hpf and 48 to 72 hpf, respectively. Atorvastatin expanded scxa:mcherry expression at the cleithrum base (arrowhead), extending distally to the actinotrichia. (J) qPCR quantification revealed that atorvastatin increased scxa expression at 56 hpf and 72 hpf upon incubation from 32 to 56 hpf and 48 to 72 hpf, respectively, compared with controls. n=4, whole embryo (32-56 hpf) and head region (48-72 hpf); Welch's two-tailed t-test. (K) qPCR quantification revealed that fluvastatin increased col1a2 expression at 72 hpf, after incubation from 32 to 48 hpf compared with controls. The combination of fluvastatin, which is characterized by a shorter half-life and a shortened exposure window (Fig. S2M-R), mitigated the toxic effects observed at 72 hpf with atorvastatin treatment. n=3, head region, Welch's two-tailed t-test. (L) Atorvastatin increased the quantity of craniofacial scxa+ cells at 56 hpf and 72 hpf after incubation from 32 to 56 hpf and 48 to 72 hpf, respectively, compared with controls. (M) Atorvastatin increased the quantity of pectoral fin scxa:mcherry+ cells at 56 hpf and 72 hpf upon incubation from 32 to 56 hpf and 48 to 72 hpf, respectively, compared with controls. (N) Atorvastatin increased the quantity of col1a2+ cells at 80 hpf, after incubation from 32 to 56 hpf compared with controls. (J,K) Data are mean±s.d. (L-N) Red bars indicate mean; individual points represent values for individual embryos; Mann–Whitney-Wilcoxon test. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. Ventral (B-E) and lateral (F-I) views, anterior towards the left.

Article Snippet: Embryos were incubated in the following compounds: atorvastatin, lovastatin, fluvastatin, FTI-277 and SU5402 (Tocris Bioscience); simvastatin, aphidicolin and GGTI-286 (Sigma-Aldrich); and RO48-8071, EHT-1864, ML-141, Y-27632 and SB-431542 (Cayman Chemical).

Techniques: Incubation, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: Current Issues in Molecular Biology

Article Title: Pro-Angiogenetic Effects of Purified Extracts from Helix aspersa during Zebrafish Development

doi: 10.3390/cimb44080232

Figure Lengend Snippet: Pretreatment with a VEGF pathway inhibitor prevented the formation of intersomitic vessels: The panels show representative images of a WISH assay performed with fli1 as an endothelial marker, at 36 hpf, in untreated ( A , C , E , G , I ) and treated embryos ( B , D , F , H , J ) with SUGEN 5416. Previously, some embryos were treated with snail derivatives via Method I and controls with fish water and 0.1% DMSO (see Materials and Methods, and ). Magnification of the trunk region (32×). Two replicates were performed ( n = 15). Ratios at the bottom-left part of each picture specify the number of embryos showing the same staining pattern, compared to the total number of embryos used for each experiment. The images were taken in lateral position at 32× magnification with a Zeiss Axiozoom V13 (Zeiss, Jena, Germany) microscope, equipped with a PlanNeoFluar Z 1×/0.25 FWD 56 mm lens and Zen Pro software.

Article Snippet: Zebrafish embryos were then treated at 4 hpf with the snail derivatives LH, LM, LH3, and LM2, and then at 15 hpf with the VEGF inhibitor SU5416 (Sunitinib, Merck KGaA, Darmstadt, Germany) at a final concentration of 5 μg/mL in fish water and incubated in 3 cm diameter well-plates.

Techniques: Marker, Staining, Microscopy, Software