Osteocyte gene expression analysis in mouse bone: Optimization of a laser-assisted microdissection protocol
| dc.rights.license | open | en_US |
| hal.structure.identifier | Bioingénierie tissulaire [BIOTIS] | |
| dc.contributor.author | PALMIER, Mathilde | |
| hal.structure.identifier | Neurocentre Magendie : Physiopathologie de la Plasticité Neuronale [U1215 Inserm - UB] | |
| dc.contributor.author | MAÎTRE, Marlène | |
| hal.structure.identifier | Neurocentre Magendie : Physiopathologie de la Plasticité Neuronale [U1215 Inserm - UB] | |
| dc.contributor.author | DOAT, Hélène | |
| hal.structure.identifier | Neurocentre Magendie : Physiopathologie de la Plasticité Neuronale [U1215 Inserm - UB] | |
| dc.contributor.author | LESTÉ-LASSERRE, Thierry | |
| hal.structure.identifier | Bioingénierie tissulaire [BIOTIS] | |
| dc.contributor.author | MAUREL, Delphine B. | |
| hal.structure.identifier | Bioingénierie tissulaire [BIOTIS] | |
| dc.contributor.author | BOIZIAU, Claudine
ORCID: 0000-0002-0475-2571 IDREF: 85228370 | |
| dc.date.accessioned | 2025-03-27T14:57:46Z | |
| dc.date.available | 2025-03-27T14:57:46Z | |
| dc.date.issued | 2024-06-24 | |
| dc.identifier.uri | https://oskar-bordeaux.fr/handle/20.500.12278/205727 | |
| dc.description.abstractEn | Among bone cells, osteocytes are the most abundant, but also the most challenging to study because they are located inside a dense mineralized matrix. Due to their involvement in bone homeostasis, diverse tools are needed to understand their roles in bone physiology and pathology. This work was aimed at establishing a laser-assisted microdissection protocol to isolate osteocytes and analyze their gene expressions. The goal was to overcome the limitations of the technique currently most used: RNA extraction from the whole bone. To perform laser microdissection and subsequent gene expression analysis, the five main steps of the protocol have been adapted for the bone tissue. After testing many parameters, we found that the best options were (1) take unfixed snap-frozen tissue, (2) cryosection with a supported tape system to improve the tissue morphology if necessary, (3) microdissect regions of interest, and (4) recover the bone pieces by catapulting, if feasible, or by gravity. Finally, RNA extraction (5) was the most efficient with a precipitation method and allowed quantifying the expression of well described osteocyte genes (Gja1/Cx43, Phex, Pdpn, Dmp1, Sost). This work describes two protocols optimized for femur and calvaria and gives an overview of the many optimization options that one could try when facing difficulties with laser microdissection. © The Author(s) 2024. | |
| dc.language.iso | EN | en_US |
| dc.subject.en | UV laser | |
| dc.subject.en | bone cells | |
| dc.subject.en | cortical bone | |
| dc.subject.en | gene expression | |
| dc.title | Osteocyte gene expression analysis in mouse bone: Optimization of a laser-assisted microdissection protocol | |
| dc.type | Article de revue | en_US |
| dc.identifier.doi | 10.1093/jbmrpl/ziae078 | en_US |
| dc.subject.hal | Sciences du Vivant [q-bio] | en_US |
| bordeaux.journal | JBMR Plus | en_US |
| bordeaux.volume | 8 | en_US |
| bordeaux.hal.laboratories | Bioingénierie Tissulaire (BioTis) - U1026 | en_US |
| bordeaux.issue | 8 | en_US |
| bordeaux.institution | Université de Bordeaux | en_US |
| bordeaux.institution | CNRS | en_US |
| bordeaux.institution | INSERM | en_US |
| bordeaux.institution | CHU de Bordeaux | en_US |
| bordeaux.institution | Institut Bergonié | en_US |
| bordeaux.peerReviewed | oui | en_US |
| bordeaux.inpress | non | en_US |
| hal.identifier | hal-05008957 | |
| hal.version | 1 | |
| hal.date.transferred | 2025-03-27T14:57:50Z | |
| hal.popular | non | en_US |
| hal.audience | Internationale | en_US |
| hal.export | true | |
| dc.rights.cc | Pas de Licence CC | en_US |
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