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Cellular Function and Structure of Primary Cilia

Mohieldin, Ashraf M.
http://rave.ohiolink.edu/etdc/view?acc_num=mco1438784302

Abstract Details

2015, Doctor of Philosophy (PhD), University of Toledo, Medicinal Chemistry.
The dysfunction of primary cilia has been linked to a list of diseases, now known as ciliopathy disorders. As a result, the primary cilia have been the focus of intense research and clinical studies for the past years. Even though many ciliary proteins of primary cilia have been identified, there are still many to be recognized. Therefore, we were interested in studying the primary cilia in depth to fully understand their function and structure. Here we report that primary cilia exhibit a swelling membrane structure, which based on their unique ultrastructure and motility, could be mechanically regulated by fluid-shear stress. In addition, we for the first time reported that monosialodihexosylganglioside (GM3S) and bicaudal-c1 (Bicc1) were members of the primary ciliary proteins localized particularly on the ciliary swelling membrane structure. In doing so, we have developed a new differential side-view imaging technique and called it “The formvar-folding technique” where cells were grown on a flexible substratum. The technique allowed us to record and measure cilioplasm calcium signals, as well as performs ciliary protein localization and immunofluorescence studies. However, the main challenge in studying the ultrastructure of primary cilia is particularly due to the high sensitivity of cilium to chemical fixation, which is required by many imaging techniques. To come over this issue, we have developed other techniques and used the combined freeze fracture transmission electron microscopy (FFTEM) and high pressure freezing (HPF) techniques to study the ultrastructure of primary cilia. We showed that the FFTEM/HPF is an inexpensive procedure that can be used to reveal the ultrastructure of primary cilia without the use of any chemical fixation and allows maintenance of the cilia morphology in its best natural form. To further investigate the ciliary membrane proteins of primary cilia, we used a sensitive multidimensional protein identification technique (MudPIT) to generate a ciliary protein database. In this study, we reported 19 new ciliary proteins. Overall, our studies provided important steps to better understand the importance of primary cilia and how they play a role in ciliopathy disorders.
Surya Nauli, PhD (Advisor)
209 p.
Biochemistry; Biology; Biomedical Research; Cellular Biology; Molecular Biology
Primary Cilia, ADPKD and Primary Cilium, Ciliary Bulb, Formvar folding techniques, Freeze Fracture of Primary cilia and Primary cilia proteome

Recommended Citations

Citations

  • Mohieldin, A. M. (2015). Cellular Function and Structure of Primary Cilia [Doctoral dissertation, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=mco1438784302

    APA Style (7th edition)

  • Mohieldin, Ashraf. Cellular Function and Structure of Primary Cilia. 2015. University of Toledo, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=mco1438784302.

    MLA Style (8th edition)

  • Mohieldin, Ashraf. "Cellular Function and Structure of Primary Cilia." Doctoral dissertation, University of Toledo, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=mco1438784302

    Chicago Manual of Style (17th edition)

mco1438784302
515
© 2015, some rights reserved.Creative Commons License Cellular Function and Structure of Primary Cilia by Ashraf M. Mohieldin is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by University of Toledo Health Science Campus and OhioLINK.