臺灣博碩士論文加值系統

Echnoid (Ed) is a cell adhesion molecule (CAM), which regulates many function durng Drosophila development. To examine how this cell adhesion molecule exerts its biological functions, several screens have been performed. One major protein was identified to be Receptor of Activated protein C Kinase 1 (RACK1). The molecular interaction between Ed and RACK1 was confirmed both in vivo and in vitro (Hsu, unpublished data). RACK1 thus raised our interests to investigate.
We obsreved that RACK1 mutant cell form rough and concaved compound eye. The RACK1 mutant photoreceptors located basally in third instar larvae eye disc. These mislocalized photoreceptors express the epithelial markers such as aPKC, apical junction proteins (Patj), Disc large (Dlg), and E-cadherin above their nucleus. These proteins are present apical to mislocalized nuclei, but a more basal localization. The apical constriction in RACK1 mutant clones of furrow was normal, and no apical constriction were observed in more posterior region. We found that RACK1 exerts its effects on eye development by regulating the accumulation of F-actin and phosphorylation of MRLC of nonmuscle myosin II. The accumulation of F-actin in ectopic clones were present both anterior and posterior to the furrow, and the excessive F-actin present in RACK1 mutant photoreceptors causes them to adopt a more basal location in the eye disc. Also the level of F-actin and phosphorylated MRLC is increased in RACK1 mutant clones were also found in wing discs. During oogenesis, RACK1 mutant follicle cell sometimes form multiple layer in later stages with decreased F-actin expression.

1. Introduction 1
2. Materials and Methods 5
2.1 Drosophila stock 5
2.2 RACK1 RNA interference 5
2.3 Mosaic Analysis and Ectopic Expression 6
2.4 Cuticle Preparation 6
2.5 Immunohistochemistry 7
3. Results 8
3.1 RACK1 Is a Highly Conserved Protein 8
3.2 Mosaic Analysis on the RACK1 P-element Insertion Line Leads to Rough and Concaved Eye Phenotype 10
3.3 RACK1EY00128 Clones Cause Mislocalization of Photoreceptor Nuclei Basally 10
3.4 RACK1EY00128 Photoreceptors Lose Their Apical Localization but Maintain Their Polarity 11
3.5 RACK1 Mediates Its Effect on Eye Through F-actin Regulation and Myosin Regulatory Light Chain Phosphorylation 13
3.6 RACK1EY00128 Clones Cause Multiple Phenotypes in Notum and Wing Margin Hair 14
3.7 RACK1 Mutant Cell Move Basally in Wing Discs 16
4. Discussion 17
4.1 The RACK1 mutant clone cause the concaved phenotype in eye disc by accumulation of F-actin and phosphprylation of MRLC 17
4.2 Drosophila RACK1 is important for PKC-binding 18
4.3 PKC mediates the phosphorylatopn of MRLC and actin activity 18
4.4 Tissue invagination requires actin meshwork 19
4.5 The RACK1 mutant follicle cell during oogenesis 20
5. Figures 21
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