(a–h) Wholemount Th immunostaining visualizing sympathetic projections from STGs in E15.5 thoraxes (a–d) and hearts (e–h) from Ednra-/- (b, f), Edn1-/- (c, g), Ece1-/- (d, h) and a control (a, e) …
(a–b) Neurite outgrowths from E14.5 control STG (a) and Ednra-/- STG (b) cocultured with thoracic descending aorta segments (dAo) dissected from littermate control embryos were visualized by …
(a) Quantitative real-time PCR analysis for class three semaphorin receptor and co-receptor genes. Total RNA was extracted from individual E14.5 STG isolated from total seven different litters, …
(a) Schematic diagram of conserved MEF2C binding sites in the Plxna4 locus. The Plxna4 locus is shown in the center with transcriptional start site (TSS, bent arrow) and exons (vertical black lines) …
(a–d) Wholemount immunostaining visualizing the dorsal surface of hearts from a 7-week-old Th-Cre; Ednrafl/+; Plxna4+/- double heterozygote (c), a Th-Cre; Ednrafl/- mutant (d), and single …
(a) A compiled representation of Th+ area on the heart dorsal surface of 6–7 week old Th-Cre; Ednrafl/+; Plxna4+/- double heterozygote (n = 7), Th-Cre; Ednrafl/- mutant (n = 4), and littermate …
(a–c) Changes in heart rate following amphetamine (a;<2 min, (b;>10 min) and isoproterenol (c) administration measured in indicated 7-week-old single heterozygotes, double heterozygotes, conditional …
(a–c) Power spectrum plots of baseline heart rate fluctuation in Th-Cre; Ednrafl/+; Plxna4+/- (b), Th-Cre; Ednrafl/- (c) mutants, and a littermate control (a). The cardiac interbeat intervals in …
(a–h) Transverse sections from E14.5 Ednra+/-; Plxna4+/- (b, f), Plxna4-/- (c, g), Ednra-/- (d, h) mutant embryos and a littermate control embryo at the level of carotid artery bifurcation (a–d) and …
(a–h) Neurite outgrowth from explanted STG co-cultured with COS cell aggregates were visualized by whole mount Th staining. STG dissected from E14.5 embryos of the genotypes indicated at left were …
(a–h) Transverse sections from an E14.5 Ednra-/- mutant (b, f, j, n) and littermate control (a, e, i, m) embryos at the thoracic levels stained with Sema3a (a–b), Sema3f (e–f), Sema6a (i–j) and …
An Ednra-expressing STG neuron located in the vicinity of distinct vascular trajectories (vein and artery) sees venous-derived endothelin as a primary attractive guidance cue and initiates axonal …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (M. musculus) | Ednrafl, ETAflox | PMID: 14585980 | MGI:2687362, RRID:IMSR_RBRC06323 | |
Genetic reagent (M. musculus) | MEF2Cfl, Mef2cloxP | PMID: 17336904 | MGI:3718323 | |
Genetic reagent (M. musculus) | Edn1-, EdnRA-, ET-1- | PMID: 8152482 | MGI:1857637 | |
Genetic reagent (M. musculus) | Ece1-, ECE-1 - | PMID: 9449665 | MGI:2158947, RRID:IMSR_RBRC06166 | |
Genetic reagent (M. musculus) | Plxna4-, PLA4 KO, PlexinA4- | PMID: 15721238 | MGI:3579185, RRID:MMRRC_030407-MU | |
Genetic reagent (M. musculus) | ThCre, Th::Cre | PMID: 1785595 | MGI:5607431, RRID:MMRRC_029177-UCD | |
Genetic reagent (M. musculus) | Wnt1Cre, Tg(Wnt1-cre/Esr1)10Rth | PMID: 9843687 | MGI:2447684 | |
Cell line (Cercopithecus aethiops) | COS-7 | ATCC | CRL-1651, RRID: CVCL_0224 | LOT: 5784927 Originally tested negative for Mycoplasma, Bacterial and fungal contamination (purchase date:6/30/2016). Cells have not independently authenticated for each passage used for experiments and data collection. Cells were not independently tested for mycoplasma contamination. |
Antibody | Sheep polyclonal anti-Tyrosine Hydroxylase | Millipore | AB1542, RRID:AB_90755 | IHC (1:400) |
Antibody | Rabbit polyclonal anti-Plexin A4 | Novus Biological | MBP1-85128, RRID:AB_11013411 | IHC (1:400) |
Antibody | Rabbit polyclonal anti-Semaphorin 3A | Abcam | ab23393, RRID: AB_447408 | IHC (1:300) |
Antibody | Rabbit polyclonal anti-Semaphorin 3F | Abcam | an203394, RRID:AB_2783521 | IHC (1:500) |
Antibody | Rabbit polyclonal anti-Semaphorin 6A | Abcam | ab72369 | IHC (1:200) |
Antibody | Mouse monoclonal anti-Semaphorin 6B | SCBT | sc-390928, RRID:AB_2783522 | IHC (1:200) |
Antibody | Rabbit polyclonal anti-HCN4 | Abcam | ab69054, RRID: AB_1861080 | IHC (1:500) |
Antibody | Mouse monoclonal anti-SYP | SCBT | sc-17750, RRID: AB_628311 | IHC (1:500) |
Recombinant DNA reagent | Ad-GFP-2A-mSema3A | VectorBio | ADV-271580 | |
Sequence-based reagent | qRT-PCR primers (hprt) | primer bank ID: 7305155a1 | ||
Sequence-based reagent | qRT-PCR primers (nrp1) | primer bank ID: 6679134a1 | ||
Sequence-based reagent | qRT-PCR primers (plxna3) | primer bank ID: 667939a1 | ||
Sequence-based reagent | qRT-PCR primers (plxna4) | primer bank ID: 28461143a1 | ||
Commercial assay or kit | RNaseOUT | Thermofisher | 10777019 | |
Commercial assay or kit | Trizol LS | Thermofisher | 10296028 | |
Commercial assay or kit | RNeasy Micro Kit | Qiagen | 74004 | |
Commercial assay or kit | Ovation RNA-Seq System V2 | NuGEN | ||
Commercial assay or kit | Encore Rapid Library System | NuGEN | ||
Chemical compound, drug | amphetamine | Sigma A5880 | ||
Chemical compound, drug | isoproterenol | Sigma I6504 | ||
Chemical compound, drug | propranolol | Sigma P0884 | ||
Software, algorithm | Fiji - Image J | https://imagej.net/Fiji | RRID:SCR_003070 | |
Software, algorithm | CASAVA (v1.8.2) | Illumina | RRID:SCR_001802 | |
Software, algorithm | tophat v1.3 | Trapnell et al. (2009) | doi:10.1093/ bioinformatics/btp120 | |
Software, algorithm | U-Seq version 8.0.2 | Nix et al. (2008) | doi:10.1186/1471 -2105-9-523 | |
Software, algorithm | LabChart 8 | AD Instruments | RRID:SCR_001620 |