Article Title: Cardiac neural crest contributes to cardiomyocytes in amniotes and heart regeneration in zebrafish
Figure Lengend Snippet: ( A ) Schematic diagram of the approach: Replication Incompetent Avian (RIA) retrovirus encoding nuclear H2B-YFP was injected into the lumen of the hindbrain from which cardiac neural crest arises. ( B ) One day post-infection (HH14), whole mount image (lateral view) showing virally labeled progeny (green) in the cardiac migration stream en route to pharyngeal arch 3. ( B’ ) Brightfield image to show anatomical information. A, anterior; P, posterior; D, dorsal; V, ventral. ( C ) Two days post-infection (HH18), virally labeled cardiac crest has populated pharyngeal arches 3, 4 and 6, highlighted with dashed line. ( D ) Transverse section showing that labeled cardiac crest expresses neural crest marker HNK-1 (red). D, dorsal; V, ventral; L, left; R, right. ( E ) Low magnification transverse section of an E6 embryo (DAPI, blue). Dashed boxes show relative positions of cardiac crest-derived populations. ( F–H ) High magnification image of selected regions in E: pharyngeal arch arteries ( F ); aorticopulmonary septum ( G ); Neural crest derivatives located in the outflow tract express Troponin T (magenta), a myocardium marker ( H, H’ ). ( I ) Temporal map of the establishment of distinct cardiac neural crest derivatives. Labeled cells initially are in the migration stream, cranial nerve IX (CN-IX) and mesenchyme around pharyngeal arch arteries (PAA). Subsequently, they populate the aorticopulmonary septum (APS), myocardium (MYO) and interventricular septum (IVS). Separate channels are displayed in . Light blue indicates known neural crest derivatives. Dark blue reflects newly discovered neural crest derivatives. Scale bars: B, C, E 400 μm; D, F, G, H 100 μm.
Article Snippet: Using a standard transfection protocol, chick DF1 cells (ATCC, Manassas, VA; #CRL-12203, Lot number 62712171, RRID: CVCL_0570 , Certificate of Analysis with negative mycoplasma testing at the ATCC website) were transfected with RIA-H2B-YFP plasmid (RRID: Addgene_96893 ) and ENV-A plasmid in 15 cm culture dishes.
Techniques: Injection, Infection, Labeling, Migration, Marker, Derivative Assay