Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations
Zebrafish (ZF, Danio rerio) has emerged as an important and popular model species to study different human diseases. Key regulators of skeletal development and calcium metabolism are highly conserved between mammals and zebrafish. The corresponding orthologs share significant sequence similarities a...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2013-05-01
|
| Series: | Frontiers in Genetics |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fgene.2013.00074/full |
| _version_ | 1818021401877020672 |
|---|---|
| author | Mohammad Jakir Hosen Mohammad Jakir Hosen Olivier M. Vanakker Andy eWillaert Ann eHuysseune Paul eCoucke Anne eDe Paepe |
| author_facet | Mohammad Jakir Hosen Mohammad Jakir Hosen Olivier M. Vanakker Andy eWillaert Ann eHuysseune Paul eCoucke Anne eDe Paepe |
| author_sort | Mohammad Jakir Hosen |
| collection | DOAJ |
| description | Zebrafish (ZF, Danio rerio) has emerged as an important and popular model species to study different human diseases. Key regulators of skeletal development and calcium metabolism are highly conserved between mammals and zebrafish. The corresponding orthologs share significant sequence similarities and an overlap in expression patterns when compared to mammals, making ZF a potential model for the study of mineralization related disorders and soft tissue mineralization. To characterize the function of early mineralization-related genes in ZF, these genes can be knocked down by injecting morpholinos (MOs) into early stage embryos. Validation of the MO needs to be performed and the concern of aspecific effects can be addressed by applying one or more independent techniques to knock down the gene of interest. Post-injection assessment of early mineralization defects can be done using general light microscopy, calcein staining, Alizarin red staining, Alizarin red-Alcian blue double staining and by the use of transgenic lines. Examination of general molecular defects can be done by performing protein and gene expression analysis, and more specific processes can be explored by investigating ectopic mineralization related mechanisms such as apoptosis and mitochondrial dysfunction. In this paper, we will discuss all details about the aforementioned techniques; shared knowledge will be very useful for the future investigation of zebrafish models for ectopic mineralization disorders and to understand the underlying pathways involved in soft tissue calcification. |
| first_indexed | 2024-04-14T08:18:19Z |
| format | Article |
| id | doaj.art-62b3d1f9a57c406c91c9ac0ee33bbe42 |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-04-14T08:18:19Z |
| publishDate | 2013-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Genetics |
| spelling | doaj.art-62b3d1f9a57c406c91c9ac0ee33bbe422022-12-22T02:04:20ZengFrontiers Media S.A.Frontiers in Genetics1664-80212013-05-01410.3389/fgene.2013.0007439247Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observationsMohammad Jakir Hosen0Mohammad Jakir Hosen1Olivier M. Vanakker2Andy eWillaert3Ann eHuysseune4Paul eCoucke5Anne eDe Paepe6Center for Medical Genetics, Ghent University HospitalShahjalal University of Science and TechnologyCenter for Medical Genetics, Ghent University HospitalCenter for Medical Genetics, Ghent University HospitalGhent UniversityCenter for Medical Genetics, Ghent University HospitalCenter for Medical Genetics, Ghent University HospitalZebrafish (ZF, Danio rerio) has emerged as an important and popular model species to study different human diseases. Key regulators of skeletal development and calcium metabolism are highly conserved between mammals and zebrafish. The corresponding orthologs share significant sequence similarities and an overlap in expression patterns when compared to mammals, making ZF a potential model for the study of mineralization related disorders and soft tissue mineralization. To characterize the function of early mineralization-related genes in ZF, these genes can be knocked down by injecting morpholinos (MOs) into early stage embryos. Validation of the MO needs to be performed and the concern of aspecific effects can be addressed by applying one or more independent techniques to knock down the gene of interest. Post-injection assessment of early mineralization defects can be done using general light microscopy, calcein staining, Alizarin red staining, Alizarin red-Alcian blue double staining and by the use of transgenic lines. Examination of general molecular defects can be done by performing protein and gene expression analysis, and more specific processes can be explored by investigating ectopic mineralization related mechanisms such as apoptosis and mitochondrial dysfunction. In this paper, we will discuss all details about the aforementioned techniques; shared knowledge will be very useful for the future investigation of zebrafish models for ectopic mineralization disorders and to understand the underlying pathways involved in soft tissue calcification.http://journal.frontiersin.org/Journal/10.3389/fgene.2013.00074/fullZebrafishembryosmorpholinomineralizationosteogenic pathways |
| spellingShingle | Mohammad Jakir Hosen Mohammad Jakir Hosen Olivier M. Vanakker Andy eWillaert Ann eHuysseune Paul eCoucke Anne eDe Paepe Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations Frontiers in Genetics Zebrafish embryos morpholino mineralization osteogenic pathways |
| title | Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations |
| title_full | Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations |
| title_fullStr | Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations |
| title_full_unstemmed | Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations |
| title_short | Zebrafish models for ectopic mineralization disorders: practical issues from morpholino design to post-injection observations |
| title_sort | zebrafish models for ectopic mineralization disorders practical issues from morpholino design to post injection observations |
| topic | Zebrafish embryos morpholino mineralization osteogenic pathways |
| url | http://journal.frontiersin.org/Journal/10.3389/fgene.2013.00074/full |
| work_keys_str_mv | AT mohammadjakirhosen zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT mohammadjakirhosen zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT oliviermvanakker zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT andyewillaert zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT annehuysseune zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT paulecoucke zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations AT anneedepaepe zebrafishmodelsforectopicmineralizationdisorderspracticalissuesfrommorpholinodesigntopostinjectionobservations |