72568 - Biochemistry of cellular signalling with laboratory

Learning outcomes

At the end of the course the students has knowledge and can discuss the main biochemical mechanisms  involved in intracellular signalling and in the functional interactions among mitochondria and other organelles under physiological and some pathological conditions.

Students also become familiar with the basic techniques for handling of the animal cells in vitro culture. In particular, they acquire the main methods for measurement of cell viability/growth and energetic efficiency and, for the evaluation of biochemical and molecular players involved in cell signalling. Finally, the student is able to: analyze and discuss the main cell biochemistry topics; identify the most effective methodologies for solving specific biological issues; perform analysis to identify alterations in cellular energy functions.

 

Course contents

Lectures

Planning of didactic activity: introduction to the following aspects of the didactic module: learning outcomes; course contents; readings/bibliography; teaching methods and tools, assessment methods; when and how to meet students (office hours).

Signaling molecular platforms: i) cell signaling features; ii) interaction and functional integration between 7TM receptors and RTKs in the integration of cell signalling: iii) Phospholipase C: molecular organisation and regulation; iv) Ca²⁺ as an universal and versatile second messenger: spatio-temporal aspects; v) methods for in vivo measurement of Ca²⁺ cytosolic levels; vi) arrestin as molecular scaffold in the receptor desensitisation, in endocytosis, and in cellular signalling.

Mitochondria and signalling transduction:

1. Ultrastructure versus network: mitochondrial cristae as an example of supramolecular organization of biological membranes; mitochondrial reticulum and its morphological and functional characteristics; mitochondrial dynamics: molecular mechanisms and proteins involved in the regulation of fusion and fission; quality control process of mitochondria or mitophagy; methodologies for the analysis of the structure and function of mitochondria.
2. Mitochondria as a cellular signalling platform: metabolites versus oncometabolites; molecular pathways activated by mitochondrial signals: AMPK/mTOR and autophagy; HIF1a and hypoxic/metabolic adaptation.
3. Signaling between mitochondria and ER: ultrastructural features of the subcellular compartment MAMs and protein networks involved in its maintenance and in its cellular function; role of MAMs in mitochondrial calcium homeostasis (transport systems, structure and function of the Ca²⁺ uniport); in vivo measurement of mitochondrial Ca²⁺ using bio-imaging techniques; MAMs as a platform for the biosynthesis of phospholipids between endoplasmic reticulum and mitochondria.

 

Experimental laboratory

The experimental laboratory will introduce students 1) to the use of cellular models for the analysis of some biochemical and molecular parameters using cell viability, SDS-Page/Western blotting and immunofluorescence assays; 2) to the analysis and discussion of the data obtained during the practical laboratory.

 

Readings/Bibliography

Necessary educational supports

Students will be provided with further recommended bibliographical references (reviews and original papers) and the protocol for experimental laboratory for a further deepening of the program contents. The educational supports will be provided as PDF files and will be available to the students on the IOL platform (https://iol.unibo.it/).

Advised educational supports

The teacher advises students to consult the following textbooks for clarifications on the basic concepts of cellular and structural biochemistry: i) “Cells” by Lewin et al; ii) “Protein Structure and Function” by Petsko and Ringe; iii) “Lehninger Principles of Biochemistry” by Nelson and Cox.

 

 

 

 

Teaching methods

The teaching method used is based on lectures during which the contents of the program will be illustrated and discussed using PowerPoint slides. Attendance to such lessons is not mandatory but it is highly recommended as the contents of the program will be presented and explained by the teacher and will be discussed with the whole class. This teaching method will facilitate the learning of contents and will allow the achievement of the knowledge and skills by the whole class. Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students.

During the experimental experience, students will be supervised by the teacher and the academic tutor in charge, with a constant dialogue and discussion aimed at clarifying the analytical approaches, the individual experiments performed and the methodologies used. The practical experience in laboratory will be carried out in single-double place at the workbench by using several and different laboratory equipment. Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students. Attendance at the laboratory is mandatory.

 

 

 

 

 

 

Assessment methods

The learning test assessment of lectures and experimental laboratory teaching will consist in an interview-test of 3-4 questions in order to verify and evaluate the student's knowledge of the contents developed and discussed during the lessons and the experimental laboratory. Moreover, the student's ability to integrate and link the different topics with particular attention to the use of scientific terminology and to the proper and accurate exposure will be evaluated. In order to obtain a final grade of 30/30 with honors, the student must show to know in depth all the topics covered during the lessons. Further, the student must explain and integrate the topics with properties of scientific language. To obtain a final grade of 30/30, the student must explain contents covered during the lessons and show the ability to properly integrate them with properties of scientific language. The final grade will be scaled from 30/30 to 18/30 based on the number of questions to which the student is able to answer and on her/his ability to integrate the topics with properties of scientific language. In particular, to obtain the minimum grade of 18/30 the student must show to have basic knowledge of all the contents discussed during the lessons and not be able to integrate them with properties of scientific language. The vote will be considered valid within the calendar year.

 

Teaching tools

The contents of the lectures and experimental laboratory will be presented using PowerPoint slides and will be discussed with the whole class through appropriate teaching supports. Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students. 

Office hours

See the website of Anna Maria Porcelli