Expression of the Wilm's Tumor Gene WT1 During Diaphragmatic Development in the Nitrofen Model for Congenital Diaphragmatic Hernia

Overview

Journal Pediatr Surg Int

Specialties General Surgery
Pediatrics

Date 2010 Nov 13

PMID 21072664

Citations 5

Authors

Jens Dingemann

Takashi Doi

Elke Ruttenstock

Prem Puri

Affiliations

Soon will be listed here.

Abstract

Purpose: The nitrofen model of congenital diaphragmatic hernia (CDH) reproduces a typical diaphragmatic defect. However, the exact pathomechanism of CDH is still unknown. The Wilm's tumor 1 gene (WT1) is crucial for diaphragmatic development. Mutations in WT1 associated with CDH have been described in humans. Additionally, WT1(-/-) mice display CDH. Furthermore, WT1 is involved in the retinoid signaling pathway, a candidate pathway for CDH. We hypothesized that diaphragmatic WT1 gene expression is downregulated during diaphragmatic development in the nitrofen CDH model.

Methods: Pregnant rats received vehicle or nitrofen on gestational day 9 (D9). Embryos were delivered on D13, D18 and D21. The pleuroperitoneal folds (PPFs) were dissected using laser capture microdissection (D13). Diaphragms of D18 and D21 were manually dissected. RNA was extracted and relative mRNA expression of WT1 was determined using real-time PCR. Immunofluorescence was performed to evaluate protein expression of WT1. Statistical significance was considered p < 0.05.

Results: Diaphragmatic mRNA expression of WT1 was significantly decreased in the nitrofen group on D13, D18 and D21. Intensity of immunofluorescencence of WT1 was markedly decreased in the CDH diaphragms on D13, D18 and D21.

Conclusion: Downregulation of diaphragmatic WT1 gene expression may impair diaphragmatic development in the nitrofen CDH model.

Citing Articles

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Conditional deletion of WT1 in the septum transversum mesenchyme causes congenital diaphragmatic hernia in mice.

Carmona R, Canete A, Cano E, Ariza L, Rojas A, Munoz-Chapuli R Elife. 2016; 5.

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The role of primary myogenic regulatory factors in the developing diaphragmatic muscle in the nitrofen-induced diaphragmatic hernia.

Dingemann J, Doi T, Ruttenstock E, Puri P Pediatr Surg Int. 2011; 27(6):579-82.

PMID: 21258934 DOI: 10.1007/s00383-010-2834-8.

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