Using a UAS-fbl-RNAi induced wing phenotype to identify functional interaction partners of Coenzyme A metabolism

Horstink, I.D. (Ineke) (2014) Using a UAS-fbl-RNAi induced wing phenotype to identify functional interaction partners of Coenzyme A metabolism. thesis, Medicine.

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Abstract

Pantothenate Kinase Associated Neurodegeneration (PKAN), a progressive neurodegenerative disorder, is caused by absence of pantothenate kinase leading to defects in Coenzyme A (CoA) synthesis. PKAN patients have a deleterious mutation in the PANK2 (Pantothenate Kinase) gene which disrupts the first step of the de novo synthesis of CoA. It is unknown if components of this pathway have functions other than in CoA synthesis. To elucidate this, animal models have been created of which the Drosophila model is the most promising. Drosophila has only one PANK gene, fumble (fbl), which is the ortholog of the human PANK2. Fumble mutants show, among other things, a neurodegenerative phenotype and shortened lifespan, reminiscent of the human condition, making it a possibly useful model to study the disease. To test the effect of fbl knockdown on individual organs/tissues, UAS-fbl-RNAi was overexpressed, under the control of several wing drivers, which leads to effects on size and morphology of the wings. Stocks stably carrying this wing driver-RNAi combination can be crossed with additional UAS-RNAi’s, mutants of other genes or UAS-overexpression constructs. Through this, genes that feed into or functionally interact with the CoA metabolism pathway could be identified. Following validation of the driver lines and phenotypic analysis of the RNAi overexpression phenotype we performed a small scale interaction study to test the usefulness of the wing phenotype. The results showed that the wing model can be used to screen for genetic interaction. We also demonstrated that by feeding Coenzyme A and/or pantethine it is possible to rescue the wing phenotype caused by absence of pantothenate kinase.

Item Type:	Thesis (Thesis)
Supervisor name:	Sibon, O.C.M.
Faculty:	Medical Sciences
Date Deposited:	25 Jun 2020 10:42
Last Modified:	25 Jun 2020 10:42
URI:	https://umcg.studenttheses.ub.rug.nl/id/eprint/384

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