Show simple item record

dc.contributor.authorDen Hartogh, Danja
dc.date.accessioned2018-09-24T20:33:52Z
dc.date.available2018-09-24T20:33:52Z
dc.identifier.urihttp://hdl.handle.net/10464/13693
dc.description.abstractIntroduction: Mechanisms directing mast cell differentiation are incompletely defined. Epigenetic modifications by promoter methylation have been identified as key modulators of locus-specific chromatin accessibility during mast cell differentiation, but the role of histone acetylation (HA) has not been explored. Resultant changes in gene accessibility support a trajectory of lineage-specific gene expression as unique cell types mature from pluripotent progenitor hematopoietic stem cells. The MAPK signaling pathway contributes to regulating differentiation and proliferation and directly influences HA modifiers in a host of contexts. We aim to measure how the MAPK signaling pathway influences histone modifiers during mast cell differentiation in vitro toward the identification of potential key contributors. Methods: Mast cell differentiation was initiated from cultures of isolated murine bone marrow and samples were collected throughout differentiation. Quantitative polymerase chain reaction (qPCR) measurement of the RNA level, western blotting assessment of the protein level, and flow cytometry assessment of cell-surface receptor phenotype were conducted for HDACs, HATs and key mast cell-specific markers and transcription factors. Results: The MAPK signaling pathway significantly affected the expression of histone acetyltransferases, histone deacetylases and histone H3 post-translational modification. The inhibition of ERK (SCH772984) differently influenced the differentiation of mast cells through increased HDAC4 and increased PCAF gene expression. This was accompanied with changes in Histone H3 acetylation (K9) and phosphorylation (S10) and increased FcεRIα surface receptor presence with ERK inhibition. The inhibition of p38 (Losmapimod) showed a reduction in mast cell differentiation through decreased mast cell specific transcription factors and enzymes, MITF, Tpsb2, Cpa3 and Cma1, while decreasing FcεRIα receptor presence on the cell surface. The inhibition of JNK (JNK-IN-8) had no effect on mast cell differentiation. Conclusion: This work demonstrates the differential and dynamic importance of the ERK and p38 MAPK signaling pathways in mast cell differentiation and suggests links between the mast cell lineage program and epigenetic modifications via HA. Activation of the p38 MAPK signaling pathway is shown to drive mast cell differentiation, while ERK activation hinders HSC mast cell differentiation.en_US
dc.language.isoengen_US
dc.publisherBrock Universityen_US
dc.subjectEpigeneticsen_US
dc.subjectMast-cellen_US
dc.subjectDifferentiationen_US
dc.subjectMAPK signalingen_US
dc.titleInfluence of mitogen-activated protein kinase (MAPK) signaling on mast cell differentiation and histone acetylation modifiersen_US
dc.typeElectronic Thesis or Dissertationen_US
dc.degree.nameM.Sc. Applied Health Sciencesen_US
dc.degree.levelMastersen_US
dc.contributor.departmentApplied Health Sciences Programen_US
dc.degree.disciplineFaculty of Applied Health Sciencesen_US
refterms.dateFOA2019-09-10T00:00:00Z


Files in this item

Thumbnail
Name:
Brock_Den_Hartogh_Danja_2018.pdf
Size:
10.30Mb
Format:
PDF
Description:
Danja Den Hartogh's Master of ...

This item appears in the following Collection(s)

Show simple item record