Access Type
Open Access Dissertation
Date of Award
January 2023
Degree Type
Dissertation
Degree Name
Ph.D.
Department
Physiology
First Advisor
Zhibing Zhang
Abstract
Cilia are broadly categorized into primary and motile cilia. The intraflagellar transport(IFT) system, composed of two large multimeric IFT complexes, is universally present in all cilia and facilitates bidirectional cargo transport. While the role of IFT in primary cilia and eukaryotic motile cilia has been demonstrated, the association between IFT and mammalian motile ciliogenesis remains largely unknown. Aim 1 of the present study is to uncover the role of IFT in mammalian motile ciliogenesis. By using a motile cilia-specific Ift140 knock-out mice, the present study demonstrates the significance of IFT140 in the formation and maintenance of cilia in the airways, fertility,and overall survival rate. Sperm flagella is a form of modified motile cilia. The interaction between mouse meiosis-expressed gene 1 (MEIG1) and Parkin co-regulated gene (PACRG) is essential for sperm formation and male fertility. PACRG recruits MEIG1 to the manchette, potentially through dynein axonemal light intermediate polypeptide 1 (DNALI1), for normal spermiogenesis. Aim 2 of the study is to target the MEIG1/PACRG/DNALI1 complex for the development of male contraceptives. In this study, it was discovered that H121 of PACRG is a crucial amino acid mediating the interaction between PACRG and MEIG1, which is essential for downstream cargo transport involved in sperm flagella formation. DNALI1 was 165 recognized to play a multi-faceted role in spermiogenesis, including being an upstream molecule of MEIG1/PACRG for manchette microtubule association during intra-manchette transport. Lead compounds with the potential to inhibit MEIG1/PACRG interaction were identified through a luciferase complementation assay. Aim 3 of the present study is to generate fluorescent knock-in mouse models to study the mechanism of IFT trafficking in male germ cells. The successful generation of MEIG1-mCherry and IFT20-mNeonGreen mice is described in this study, highlighting their characteristics as tools to study real-time IFT trafficking.
Recommended Citation
Yap, Yi Tian, "Regulation Of Motile Ciliogenesis And The Targeting Of Meig1/pacrg/dnali1 Complex For The Development Of Male Contraceptive" (2023). Wayne State University Dissertations. 3899.
https://digitalcommons.wayne.edu/oa_dissertations/3899