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Exosomes role in muscle communication
Università degli studi di Chieti
Dept Neuroscience Imaging and Clinical Sciences University of Chieti “G. d’Annunzio” 31, Via dei Vestini – 66013 Chieti, Italy
Type of Research Project
- Basic science
What is the background of the project?
The cells are now well established that release peptides, messenger RiboNucleic Acids (mRNAs), microRNAs (miRNAs), nucleotides and more through small vesicles including exosomes. They are involved in cell–cell communication and the transmission of pathological states. Exosomes, novel nanovesicles, are secreted by cells and released in the extracellular melieu where they could keep in contact with the surrounding cells or could be captured into the flowing blood; in this manner, their content could influence different tissues as physiological mediators and/or pathological inductors. The communication by extracellular vesicles in muscle has received a great attention due to their importance as intercellular mediators bearing proteins, lipids, antioxidants, RNAs, and microRNAs through the entire body.
What is the aim of the project?
The aim of this project is to characterize in the skeletal muscle and its myogenic stem cells, cell-cell communications, cell signaling, interactions with the environment by exosome analysis.
What techniques and methods are used?
Muscle stem cells, also called satellite cells (SCs), will be isolated by muscle biopsies with explant technique and will be cultured for amplification, in order to obtain a sufficient number. The cells will be cultured in growth and /or differentiation conditions. Exosomes released from these cell cultures will be isolated by commercial kits and characterized by western blotting analysis and then RNA extraction will be performed, using the Total Exosome RNA and Protein Isolation Kit (Invitrogen) following manufacturer’s instructions. Then, the expression of muscle specific miRNAs (myomiRs) (miR-133a and miR-133b, miR-1, miR-206, miR-16 Endogen) will be investigated with Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). GraphPad prism, ImageJ, FlowJo and Microsoft Excel softwares will be used to analyze data.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
During the period of stay in the laboratory, the student will follow all the procedures relating to the sterilization of the material necessary for cell culture procedures, the preparation of culture media and the maintenance of cell cultures, including soil changes, transplants and cell counts. The student will learn to use bench top centrifuges in order to pellet cells and separate the supernatant. The student will follow procedures for determining the amount of protein, extraction of RNA and subsequent electrophoresis and RT-PCR. The student will work with the tutor on the data analysis, and will learn how to use GraphPad prism, ImageJ, FlowJo and Microsoft Excel.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
There will be theroretical teaching provided by the tutors for the students through their participation to educative occasions such as laboratory meeting, once a week, in which will be discussed data and papers recently published.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Knowledge of cell biology and theoretical bases of molecular biology
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- James R. Edgar; What are exosomes; exactly?; BMC Biology (2016)
- Pietrangelo; Di Filippo; Locatelli; Piacenza; Farina; Pavoni; Di Donato; Innosa; Provinciali; Fulle; Extracellular Guanosine 5′-Triphosphate Induces Human Muscle Satellite Cells to Release Exosomes Stuffed With Guanosine; Frontiers in Pharmacology (2018)
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