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Published on: December 1, 2015
Detection of apoptosis-inducing factor in involuting mammary tissue by immunoelectron microscopy
M Colitti1, R Musetti, B Stefanon
1Department of Scienze della Produzione Animale, University of Udine, via delle Scienze, Udine 208-33100, Italy. monica.colitti@dspa.uniud.it
Insights
Apoptosis-inducing factor (AIF) was found in the mitochondria, cytoplasm, and nucleus of bovine mammary cells during involution. This suggests AIF moves within the cell during programmed cell death (apoptosis).
Area of Science:
- Cell Biology
- Molecular Biology
- Veterinary Science
Background:
- Apoptosis-inducing factor (AIF) is a key protein involved in programmed cell death.
- Understanding the subcellular localization of AIF is crucial for elucidating its role in apoptosis.
Purpose of the Study:
- To investigate the subcellular localization of apoptosis-inducing factor (AIF) in involuting bovine mammary tissue.
- To determine if AIF undergoes translocation within mammary epithelial cells during apoptosis.
Main Methods:
- Immunoelectron microscopy was employed to visualize AIF.
- A specific polyclonal antibody against bovine AIF was used.
- Secondary antibodies conjugated to colloidal gold particles enabled detection.
Main Results:
- AIF was specifically localized in the mitochondria, cytoplasm, and nucleus of mammary epithelial cells.
- The observed localization suggests a dynamic movement of AIF within the cell.
Conclusions:
- AIF exhibits specific subcellular localization patterns in involuting bovine mammary tissue.
- These findings support the hypothesis of AIF translocation during apoptosis in these cells.
Abstract:
Immunoelectron microscopy was used to study the subcellular localisation of apoptosis-inducing factor (AIF) in involuting bovine mammary tissue. AIF was detected using a polyclonal antibody and secondary anti-rabbit antibody conjugated to colloidal gold particles under optimised conditions. The polyclonal antibody appeared specific for the bovine antigen and immunocytological examination identified specific localisation of AIF in mitochondria, cytoplasm and nucleus of mammary epithelial cells in involuting mammary tissue, suggesting a subcellular translocation in cells undergoing apoptosis.
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