Increased fibronectin expression in developing embryos is associated with abnormal notochord in the Adriamycin rat model.
BACKGROUND
The VACTERL association is a spectrum of clinical conditions, including esophageal atresia (EA) and tracheoesophageal fistula (TEF), which affects approximately 1 in 5,000 live human births. The administration of intraperitoneal Adriamycin to pregnant rats reliably induces anomalies, such as EA and TEF, in their offspring, in what is known as the Adriamycin rat model (ARM). In affected embryos the presence of gross notochord abnormalities is commonly found, with typical features being ectopic ventral branches and adherence of the notochord to the foregut. Fibronectin (FN) is an extracellular matrix (ECM) glycoprotein present on most cell surfaces, in extracellular fluids and in plasma. FN is involved in various functions, including cell adhesion, cell motility and wound healing. Previous studies in rats have shown that a single dose of Adriamycin can produce an appreciable rise in FN levels in various organs such as kidney and heart. We hypothesised that Adriamycin administration could promote upregulation of FN expression contributing to increased gut-notochord adherence and the development of abnormal ventral notochordal branching in the ARM. This study was designed to investigate FN expression in ARM embryos.


METHODS
Adriamycin (1.75 mg/kg) was administered intraperitoneally to pregnant rats on days 7,8 and 9 of gestation (E7, E8 and E9 respectively). Control animals were given saline. Embryos recovered on E10-E14 were fixed, embedded in paraffin and sectioned. Immunohistochemistry using an anti-FN rabbit polyclonal antibody was performed.


RESULTS
FN expression in both Adriamycin and control embryos on E10, E11 and E12 was comparable. However, the levels of FN expression in Adriamycin embryos on E13 and E14 were significantly greater in embryos with abnormal notochords than in equivalent control embryos.


CONCLUSIONS
Adriamycin-induced increased expression of FN, in the ARM, may contribute to abnormal notochord development leading to the VACTERL association.
Mortell
A
A
The Children's Research Centre, Our Lady's Hospital for Sick Children, Crumlin, Dublin 12, Ireland.
O'Donnell
A M
AM
Giles
S
S
Bannigan
J
J
Puri
P
P
D000014:Abnormalities, Drug-Induced / Q000378:metabolism / Q000473:pathology; D000818:Animals; D004065:Digestive System Abnormalities / Q000139:chemically induced / Q000378:metabolism*; D004317:Doxorubicin / Q000009:adverse effects; D004622:Embryo, Mammalian / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis*; D023421:Models, Animal; D009672:Notochord / Q000002:abnormalities*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D013723:Teratogens / Q000494:pharmacology; D013997:Time Factors
D005353:Fibronectins; D013723:Teratogens; D004317:Doxorubicin