© 2003 American Thoracic Society DOI: 10.1165/rcmb.4877 Antisense Oligodeoxynucleotides Decrease LGL1 mRNA and Protein Levels and Inhibit Branching Morphogenesis in Fetal Rat LungLung Biology Research, Research Institute, The Hospital for Sick Children; Department of Physiology, University of Toronto; Departments of Pediatrics and Human Genetics, McGill University-Montreal Children's Hospital Research Institute, McGill University; and Department of Paediatrics, University of Toronto, Toronto, Ontario, Canada Address correspondence to: Neil B. Sweezey, Lung Biology Research, Research Institute, The Hospital for Sick Children, 555 University Ave., Toronto, ON, M5G 1X8 Canada. E-mail: neil.sweezey{at}sickkids.ca
We previously described the cloning of the late gestation lung 1 gene (LGL1), a novel glucocorticoid-inducible gene expressed in the mesenchyme of fetal lung. We report here evidence for a role of the LGL1 gene product (lgl1) in fetal rat lung airway branching morphogenesis, temporal and spatial localization of LGL1 mRNA and lgl1 protein in fetal rat lung, and a correction of the previously published LGL1 sequence. Both the mRNA and protein were detected during fetal lung development. LGL1 mRNA was detected from gestational Day 12 by reverse transcriptase–polymerase chain reaction, and from Day 13 by in situ hybridization. lgl1 protein was detected from Day 18 by Western analysis and from Day 16 by immunohistochemistry. The types of cells expressing LGL1 mRNA and lgl1 protein were assessed by immunohistochemical staining of adjacent serial tissue sections for markers of mesenchymal (vimentin) and smooth muscle (
Abbreviations: cysteine-rich secreted protein, CRISP • normal goat serum, NGS • oligodeoxynucleotide, ODN • phosphate-buffered saline, PBS • reverse transcriptase–polymerase chain reaction, RT-PCR This article has been cited by other articles:
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-actin) cells. As gestation advanced, increasing amounts of mRNA and protein were expressed in these cells. In support of a role for lgl1 in airway branching morphogenesis, antisense (but neither sense nor scrambled) oligodeoxynucleotides directed against LGL1 inhibited airway branching in fetal rat lung buds in explant culture, in a dose- and time-dependent manner. The levels of lgl1 protein and LGL1 mRNA expression were decreased in those explants that had inhibited airway branching, compared with the uninhibited controls. Our findings suggest that lgl1 plays an important role in fetal airway branching morphogenesis. 
