Wijesinghe et al 2014 Presentation by Jeetika Sainani Wound Healing A process we experience quite frequently but rarely understand What is this process Why does it matter Four stages of wound healing ID: 916986
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Slide1
Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts.
Wijesinghe
et. al. 2014.
Presentation by
Jeetika
Sainani
Slide2Wound Healing: A process we experience quite frequently, but rarely understand
What is this process? Why does it matter?
Four stages of wound healing
Efficiently cure woundFibroblastsFibroblastsWhy is migration so important? Disorders
Slide3How do fibroblasts receive the signal that promotes their migration?
CERK
Ceramide
C1P
C1P
cPLA
2
Time that cPLA
2
is active increases when bound
Fibroblast
AA
Metabolized
enzymatically
eicosanoids
Eicosanoids can now bind to fibroblasts
Slide4What does the effect of ablating ceramide kinase have on the release of arachidonic acid by fibroblasts?
Experiment: Measurement of arachidonic acid release in response to mechanical trauma
Mice were used to conduct experiments
Measurement taken by Scintillation
Slide5Mouse Embryonic Fibroblasts
Genetic ablation
CERK-null MEFs
CERK wild-type MEFsPrimary fibroblasts vs. immortalized fibroblastsWhat’s the difference?Fibroblasts seeded with DMEM, 10% FBS and penicillin/streptomycin3H Arachidonic Acid
Fig. 5C: Fibroblast migration patterns from immortalized ceramide kinase of both wild type and ablated fibroblasts.
Fig. 5D: Fibroblast migration patterns from primary ceramide kinase of both wild type and ablated fibroblasts.
Slide6CERK-null and CERK wild-type fibroblasts demonstrated significant differences in the release of Arachidonic Acid
Slide7References
Wijesinghe
, D. S., Brentnall, M.,
Mietla, J. A., Hoeferlin, L. A., Diegelmann, R. F., Boise, L. H., & Chalfant, C. E. (2014). Ceramide kinase is required for a normal eicosanoid response and the subsequent orderly migration of fibroblasts. Journal of Lipid Research,55(7), 1298-1309. doi:10.1194/jlr.m048207
http://www.ntanet.net/how-do-sodium-iodide-scintillation-detectors-work
https://www.biodermis.com/what-are-the-stages-of-wound-healing-s/221.htm
https://courses.washington.edu/conj/membrane/arachidonic.htm
http://lipidlibrary.aocs.org/Biochemistry/content.cfm?ItemNumber=39190
Slide8Scintillation Counter – using flashes of light to count molecules
Radioactive compound: Trituum-3