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Title: MORPHOLOGY OF ANNEXIN-V POSITIVE NEUTROPHILS FROM BOVINE BLOOD

Author
item VAN OOSTVELDT, K - U GHENT BELGIUM
item BURVENICH, C - U GHENT BELGIUM
item DOSOGNE, H - U GHENT BELGIUM
item Paape, Max

Submitted to: International Congress On Apoptosis
Publication Type: Proceedings
Publication Acceptance Date: 8/21/2001
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Apoptosis of bovine neutrophils (PMN) was studied using a double labeling technique with fluorescein isothiocyanate labeled annexin-V and propidium iodide (PI). Three different subpopulations of bovine PMN could be detected: the apoptotic cells, the necrotic cells and the living cells. The three subpopulations were sorted out and studied for morphology using flow cytometry and light microscopy. Flow cytometric analysis indicated that the apoptotic subpopulation had decreased side scatter compared to the scatter pattern of the living PMN indicating degranulation of the cells. The necrotic subpopulation showed both a decreased side scatter and a strongly decreased forward scatter indicating degranulation and shrinkage of the cells. Microscopic examination of cytocentrifuge preparations of the PMN populations indicated that the annexin-V positive cells had typical features for early apoptosis such as coalescence of nuclear lobes and the PI positive cells had typical features of necrosis with ruptured cytoplasm membrane. Results indicate that annexin-V in combination with PI is a convenient marker for studying apoptosis and necrosis of PMN, and that the flow cytometric procedure used in this study should be useful for detecting apoptosis and necrosis of bovine PMN .