
Cell Death and Injury
Code
12107
Academic unit
Faculdade de Ciências e Tecnologia
Department
Departamento de Ciências da Vida
Credits
3.0
Teacher in charge
Margarida Casal Ribeiro Castro Caldas Braga
Weekly hours
3
Total hours
34
Teaching language
Português
Objectives
In this Course we intend to deepen the study of the cellular and molecular processes involved in the different types of cell death, and in the cellular response to the injury. In parallel we will address methods of evaluation of different stages of cell death, as well as development of drugs and research of therapeutic targets.
It is expected that at the end of this course students will have achieved the following general competences: i) understanding the concept of oxidative stress; ii) to understand, through practical cases, the concept of cellular injury and the mechanisms activated in the endogenous response to the injury; iii) enumerate and distinguish different types and cell death, relating the inductors and mechanisms involved; iv) to identify the deregulation of the mechanisms involved in cell death as being the basis of several diseases; v) to use the acquired concepts and to understand strategies and therapeutic targets to modulate cell death in the different diseases; vi) acquisition of research skills of recent literature on subjects taught, in order to expose and critically discuss scientific works in the area; Vii) to acquire laboratory skills and contact with methodologies currently used in the evaluation of different injury and cell death parameters.
Subject matter
Theoretical classes
1. Oxidative stress: inducers, mechanisms involved and adaptation of cells and tissues to oxidative stress
2. Cell death in homeostasis and deregulation of cell death in pathological processes: necrosis, apoptosis, necroptosis and other forms of cell death.
2.1. General mechanisms of activation of the cell death process
2.2. Molecular mechanisms involved in the process of cell death
2.3. Cell organelles as modulators and targets of the processes involved in cell death
2.4. Cellular response to injury and reversibility of the cell death process
2.5. Deregulation of cell death in several pathologies: cancer, autoimmune diseases and infections
2.6. Deregulation of cell death in several pathologies: degenerative diseases.
3. Role of autophagy in the regulation of cell death.
3.1. Cell organelles as modulators and targets of the processes involved in autophagy.
4. Modulation of cell death. Development of drugs and research of therapeutic targets.
Practical classes:
1. Evaluation of oxidative stress
1.1. Griess reaction
1.2. Evaluation of ROS: staining with DCF-DA dye; fluorescence microscopy
2. Evaluation of cell death (apoptosis/necrosis)
2.1. DNA laddering
2.2. Evaluation of caspases activity using exogenous substract
2.3. Evaluation of nuclear morphology with Hoechst staining
2.4. Evaluation of lactate desidrogenase activity (LDH assay)
2.5. Analysis of apoptosis/necrosis by FACS: staining with annexin A5 and propidium iodide
2.6. Analysis of apoptosis/necrosis by FACS: acquisition and data analysis
Bibliography
Molecular Cell Biology. Lodish et al. 7th Ed. WH Freeman & Company, NY 2012
Molecular Biology of the Cell. Alberts et al. 5th Ed. Garland Science, NY 2007
Scientific papers
Teaching method
Theoretical classes are based on textbooks and recent articles from the scientific literature. The classes are generally of the exhibition type, with recourse to the data show. The practical classes include experimental tasks to evaluate different parameters involved in the processes of oxidative stress and/or cell death. Emphasis will be given to the analysis, interpretation and joint discussion of the results, so that the students can evaluate the contribution of the different processes studied in cell death. This knowledge should contribute to understand the study models used, as well as the evolution in the research of new drugs and new therapeutic targets.
In the continuous assessment part, students will be evaluated by their performance in the practical classes, as well as by the ability to understand and interpret the results obtained in the form of a mini-scientific article. The evaluation of the theoretical component will be done through tests and / or final exam.