Analysis of a lung defect in autophagy-deficient mouse strains

• Published in: Autophagy 2014-01, Vol.10 (1), p.45-56
• Main Authors: Cheong, Heesun, Wu, Junmin, Gonzales, Linda K, Guttentag, Susan H, Thompson, Craig B, Lindsten, Tullia
• Format: Article
• Language: English
• Subjects: Alveolar Epithelial Cells - metabolism; Alveolar Epithelial Cells - pathology; Alveolar Epithelial Cells - ultrastructure; Animals; Animals, Newborn; Atg5 KO mice; Autophagy; Autophagy-Related Protein 5; Autophagy-Related Protein-1 Homolog; Basic Research Paper; glycogen; Glycogen - metabolism; Immunohistochemistry; Lung - embryology; Lung - pathology; Lung - ultrastructure; lung development; Mice; Mice, Knockout; Microtubule-Associated Proteins - deficiency; Microtubule-Associated Proteins - metabolism; perinatal mortality; Phospholipids - metabolism; Protein-Serine-Threonine Kinases - deficiency; Protein-Serine-Threonine Kinases - metabolism; Staining and Labeling; Ulk1/2 DKO mice
• ISSN: 1554-8627; 1554-8635
• DOI: 10.4161/auto.26505

Yeast Atg1 initiates autophagy in response to nutrient limitation. The Ulk gene family encompasses the mammalian orthologs of yeast ATG1. We created mice deficient for both Ulk1 and Ulk2 and found that the mice die within 24 h of birth. When found alive, pups exhibited signs of respiratory distress. Histological sections of lungs of the Ulk1/2 DKO pups showed reduced airspaces with thickened septae. A similar defect was seen in Atg5-deficient pups as both Ulk1/2 DKO and Atg5 KO lungs show numerous glycogen-laden alveolar type II cells by electron microscopy, PAS staining, and increased levels of glycogen in lung homogenates. No abnormalities were noted in expression of genes encoding surfactant proteins but the ability to incorporate exogenous choline into phosphatidylcholine, the major phospholipid component of surfactant, was increased in comparison to controls. Despite this, there was a trend for total phospholipid levels in lung tissue to be lower in Ulk1/2 DKO and Atg5 KO compared with controls. Autophagy was abundant in lung epithelial cells from wild-type mice, but lacking in Atg5 KO and Ulk1/2 DKO mice at P1. Analysis of the autophagy signaling pathway showed the existence of a negative feedback loop between the ULK1 and 2 and MTORC1 and 2, in lung tissue. In the absence of autophagy, alveolar epithelial cells are unable to mobilize internal glycogen stores independently of surfactant maturation. Together, the data suggested that autophagy plays a vital role in lung structural maturation in support of perinatal adaptation to air breathing.