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(Concepcion) Cellular and molecular characterization of bone regeneration after traumatic injury of the amphibian larval skull
Universidad de Concepcion
Department of Cell Biology, Faculty of Biological Sciences, University of Concepcion
Dr. Hernan Montecinos
Dr. Sylvain Marcellini
Type of Research Project
- Basic science
What is the background of the project?
Our laboratory is mainly interested by the control of gene expression during amphibian osteogenesis. This research proposal pretends to extend the knowledge we have obtained during normal bone development towards the bone regeneration field. By using in vivo fluorescent dyes, we have already gathered excellent preliminary results showing that amphibian larval skull completely regenerate from a traumatic injury after 2 to 3 weeks. The present project will allow us to describe this phenomenon in greater depth, by generating new histological and gene expression data. In the long term, we hope to use this system to screen for molecular compounds that can help accelerate the regeneration process.
What is the aim of the project?
The aim of the project will be to characterize how the amphibian larval skull recovers from a traumatic injury. For this purpose we will descibe the regeneration process from a cellular and molecular perspective, by examining changes in histology and gene expression patterns, respectively
What techniques and methods are used?
Tadpoles will be anesthetized with MS222. Bone damage will be performed with a micro-drill (0,5mm in diameter). In vivo bone labelling of larvae of the amphibian Xenopus tropicalis will be performed before and after regeneration with concentrated solution of Alizarin Red and Calcein to discriminate “old” from “new” bone by fluorescent microscopy. Samples will be fixed in PFA 4%, embedded in paraffin, cut at 7 micrometers and examined by classical histology of by in situ hybridization with Dig- labelled RNA-probes to detect the expression of key skeletal genes such as collagen 1a1, osteocalcin and bone sialoprotein.
What is the role of the student?
- 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?
The student is expected to be motivated, organized, and to have a responsible attitude from a professional, bioethical and ethical perspective. The work will be performed with other members of the group, and a friendly and collaborative attitude is expected from the student. Results will also have to be clearly annotated in a Word file which will be examined on a weekly basis with the tutor. Results will be property of the LADE laboratory and of the University of Concepcion, and should not be divulgated to a third party without the tutor´s consent. The student will present his results at our lab meetings.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Yes, students will be given a brief lecture on osteogenesis and will have to read articles and reviews of interest.
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 presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic cell biology knowledge is absolutely required. There are no legal limitations. No report will be demanded, but it will be obligatory to register all the experiments, procedures and results in a Word file. Language(s): Required: English OR Spanish Accepted: English, Spanish, French
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) - Pre-Medical students from the American-British system
- Articles on amphibian regeneration: Godwin J. The promise of perfect adult tissue repair and regeneration in mammals: Learning from regenerative amphibians and fish. Bioessays. 2014 Sep;36(9):861-71.
- Skull regeneration after traumatic injury in fish and mammals: Geurtzen K. et al;. Mature osteoblasts dedifferentiate in response to traumatic bone injury in the zebrafish fin and skull. Development. 2014 Jun;141(11):2225-34. Kitami M. et al;. Prolonged Survival of Transplanted Osteoblastic Cells Does Not Directly Accelerate the Healing of Calvarial Bone Defects. J Cell Physiol. 2016 Sep;231(9):1974-82.
- Fritz A; Bertin A; Hanna P; Nualart F; Marcellini S. A single chance to contact multiple targets: distinct osteocyte morphotypes shed light on the cellular mechanism ensuring the robust formation of osteocytic networks. (2016) Journal of Experimental Zoology Part B: Molecular and Developmental Evolution; 326(5):280-9.
- Bertin A; Hanna P; Otarola G; Fritz A; Henriquez JP; Marcellini S. Cellular and molecular characterization of a novel primary osteoblast culture from the vertebrate model organism Xenopus tropicalis. (2015) Histochemistry and Cell Biology;143(4):431-42.
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