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Region-Specific Quantitation of Elastin in Fibrotic Mouse* FREE TO VIEW

Barry Starcher, PhD; C. Kuhn, MD
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*From the University of Texas Health Center (Dr. Starcher), Tyler, TX; and Memorial Hospital (Dr. Kuhn), Pawtucket, RI.

Correspondence to: Barry Starcher, PhD, Department of Biochemistry, University of Texas Health Center at Tyler, PO Box 2003, Highway 271 at Highway 155, Tyler, TX 75710-2003

Chest. 2001;120(1_suppl):S57-S58. doi:10.1378/chest.120.1_suppl.S57-a
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This study describes a method to quantitate elastin from specific regions of the lung while at the same time makes histologic or morphologic measurements of the same region. The examples used were control lungs and lungs from mice treated with bleomycin or bleomycin plus retinoic acid as fibrotic models, and Tsk mice with a genetic defect that resembles emphysema. A 3-cm biopsy punch was removed from paraffin-embedded lungs and reembedded with the plural side facing down, providing enface sectioning. Eleven 10-μm sections were removed and the last section used for histology, and the remaining 10 sections were placed in a microfuge tube. This process was repeated a total of 12 times. The histology sections were stained with Harts elastic stain, and the area was calculated by scanning the slides into PhotoShop. Each histology section was viewed and compartmentalized anatomically. The paraffin was removed from the pooled sections with xylene, and the tissue was hydrolyzed and the desmosines determined by radioimmunoassay. Normal lungs have a remarkably consistent elastin content per unit area from the pleura to the center of the lung (30 to 45 pm/mm3). Fibrotic lungs can show a dramatic increase in elastin per unit volume (100 to 150 pm/mm3), which again is consistent through the whole core of lung examined. No differences were observed in the degree of fibrosis between mice that were administered bleomycin and those receiving bleomycin plus retinoic acid. Lungs from Tsk mice have less than half the elastin per unit volume as control mice in the alveolar region (11 to 13 pm/mm3), and this increases slightly in the conducting airways region (19 to 21 pm/mm3).




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