MilliporeSigma

The role of ephrins-B1 and -B2 during fetal rat lung development.

Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (2014-12-31)
Francisca O Peixoto, Patrícia Pereira-Terra, Rute S Moura, Emanuel Carvalho-Dias, Jorge Correia-Pinto, Cristina Nogueira-Silva
ABSTRACT

BACKGROUND/ AIMS: The knowledge of the molecular network that governs fetal lung branching is an essential step towards the discovery of novel therapeutic targets against pulmonary pathologies. Lung consists of two highly branched systems: airways and vasculature. Ephrins and its receptors, Eph, have been implicated in cardiovascular development, angiogenesis and vascular remodeling. This study aims to clarify the role of these factors during lung morphogenesis. Ephrins-B1, -B2 and receptor EphB4 expression pattern was assessed in fetal rat lungs between 15.5 and 21.5 days post-conception, by immunohistochemistry. Fetal rat lungs were harvested at 13.5 dpc, cultured during 4 days and treated with increasing doses of ephrins-B1 and -B2 and the activity of key signaling pathways was assessed. Ephrin-B1 presents mesenchymal expression, whereas ephrin-B2 and its receptor EphB4 were expressed by the epithelium. Both ephrins stimulated pulmonary branching. Moreover, while ephrin-B1 did not affect the pathways studied, ephrin-B2 supplementation decreased activity of JNK, ERK and STAT. This study characterizes the expression pattern of ephrins-B1, -B2 and EphB4 receptor throughout rat lung development. Our data highlight a possible role of ephrins as molecular stimulators of lung morphogenesis. Moreover, it supports the idea that classical vascular factors might play a role as airway growth promoters.

MATERIALS
Product Number
Brand
Product Description

Supelco
Acrylamide solution, 40% in H2O, for molecular biology
Sigma-Aldrich
Acrylamide solution, 40%, suitable for electrophoresis, sterile-filtered
Sigma-Aldrich
3,3′-Diaminobenzidine tetrahydrochloride, tablet, 10 mg substrate per tablet
USP
Ascorbic acid, United States Pharmacopeia (USP) Reference Standard
Supelco
Ascorbic Acid, Pharmaceutical Secondary Standard; Certified Reference Material
Ascorbic acid, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
Sigma-Aldrich
L-Ascorbic acid, BioXtra, ≥99.0%, crystalline
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99%
Sigma-Aldrich
L-Ascorbic acid, reagent grade, crystalline
Sigma-Aldrich
Acrylamide, for molecular biology, ≥99% (HPLC)
Sigma-Aldrich
L-Ascorbic acid, meets USP testing specifications
Sigma-Aldrich
L-Ascorbic acid, reagent grade
Sigma-Aldrich
L-Ascorbic acid, suitable for plant cell culture
Sigma-Aldrich
L-Ascorbic acid, suitable for cell culture, suitable for plant cell culture, ≥98%
Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ACS reagent, reag. ISO, Ph. Eur., 99.7-100.5% (oxidimetric)
Supelco
Hydrogen peroxide solution, ≥30%, for trace analysis
Sigma-Aldrich
Hydrogen peroxide solution, 34.5-36.5%
Sigma-Aldrich
L-Ascorbic acid, BioUltra, ≥99.5% (RT)
Millipore
Hydrogen peroxide solution, 3%, suitable for microbiology
Sigma-Aldrich
Acrylamide, purum, ≥98.0% (GC)
Sigma-Aldrich
Acrylamide, for Northern and Southern blotting, powder blend
Sigma-Aldrich
L-Ascorbic acid, tested according to Ph. Eur.
Sigma-Aldrich
Hydrogen peroxide solution, 30 % (w/w) in H2O, contains stabilizer
Sigma-Aldrich
L-Ascorbic acid, ACS reagent, ≥99%
Sigma-Aldrich
L-Ascorbic acid, FCC, FG
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ≥99.0% (RT)
Sigma-Aldrich
Hydrogen peroxide solution, contains potassium stannate as inhibitor, 30-32 wt. % in water, semiconductor grade, 99.999% trace metals basis
Supelco
L-Ascorbic acid, analytical standard