Thrombospondin (TSP) is a secreted multidomain glycoprotein that was first discovered in platelets. The TSP family consists of five extracellular calcium binding proteins, TSP-1 through -5. TSP-1 has multiple functions including adhesion, proliferation, migration, angiogenesis and apoptosis. Recently, it has also been shown to regulate inflammation and is released during an acute phase of inflammation. In an animal model of acute lung injury (ALI), Rane laboratory demonstrated that TSP-1 was released in bronchoalveolar lavage fluid (BALf). It was also shown that Baclofen treatment of these animals was protective with concurrent inhibition of TSP-1 release. These findings led to the question, does TSP-1contribute to ALI? The goal of this research project was to isolate TSP-1 cDNA from monocytes and to subclone it into a pRSET vector and express and purify the TSP-1 protein. A complete TSP-1 gene was isolated; however, a truncated TSP-1 protein was purified as it may have degraded during purification. In the future, the purified TSP-1 protein will be administered intratracheally to rats to determine the effects of its administration on neutrophil recruitment in the BALf. Subsequent lung damage will be assessed by vascular leakage, H&E staining and TUNEL assays.
Semester/Year of Award
Marcia M. Pierce
Mentor Department Affiliation
Restricted Access Thesis
Von Handorf, Christina, "Reverse Transcription PCR, Subcloning, and Bacterial Expression of Human Thrombospondin-1 (TSP-1)" (2013). Honors Theses. 75.