Fluorescence imaging of Drosophila melanogaster tracheal system : investigating the morphology of LUBEL mutant flies
Benoit, Sabrina (2020)
Benoit, Sabrina
Åbo Akademi
2020
Julkaisu on tekijänoikeussäännösten alainen. Teosta voi lukea ja tulostaa henkilökohtaista käyttöä varten. Käyttö kaupallisiin tarkoituksiin on kielletty.
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi-fe2020070146589
https://urn.fi/URN:NBN:fi-fe2020070146589
Tiivistelmä
Ubiquitination is a post-translational modification that is a key regulator in many pathways in the cell. Mutant LUBEL (lubelMi), linear ubiquitin E3 ligase, flies are unable to form methionine 1 (M1) linked ubiquitin chains in Drosophila and die in hypoxic conditions, whereas wild-type Cantons flies survive. The Drosophila respiratory system is examined to determine if the flies are asphyxiating due to morphological changes in the tracheal epithelial tissue. To image morphological changes in the epithelial tissue, I used chitin-specific and tubulin-specific silicone rhodamine (SiR) probes and Calcofluor White staining in combination with Zeiss LSM 880 with AiryScan microscopy techniques to evaluate the respiratory system in CantonS in basal conditions, and, thus, develop a criterion for comparison with lubelMi flies. I then used STED microscopy to image taenidia within the tracheoles and measure tracheole diameter in CantonS and lubelMi flies. A novel phenotypic abnormality was seen in LUBEL mutant flies, lubelMi and lubelRNAi flies, in comparison to the control group. lubelMi and lubelRNAi flies have discontinuous, abnormal tubulin projecting from the trachea and tracheoles and measurably thicker dorsal trunks in comparison to the control group. This abnormal tubulin phenotype and increased dorsal trunk diameter could be caused by a LUBEL-influenced developmental error during organogenesis in Drosophila melanogaster larvae.
Kokoelmat
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