Date Approved

8-2018

Document Type

Dissertation

Degree Name

PhD in Cell & Molecular Biology

Department

Molecular Biology

College

Graduate School of Biomedical Sciences

First Advisor

Eric Moss, PhD

Committee Member 1

Jeremy Francis, PhD

Committee Member 2

Ronald Ellis, PhD

Committee Member 3

Hristo Houbaviy, PhD

Committee Member 4

Nathaniel Hartman, PhD

Subject(s)

RNA-Binding Proteins, Mammals, Neurogenesis, Cell Physiological Phenomena, Neurobiology

Disciplines

Cell Biology | Cellular and Molecular Physiology | Laboratory and Basic Science Research | Life Sciences | Medical Cell Biology | Medical Neurobiology | Medicine and Health Sciences | Molecular and Cellular Neuroscience | Molecular Biology

Abstract

Developmental timing is a key aspect of tissue and organ formation in which distinct cell types are generated through a series of steps from common progenitors. These progenitors undergo specific changes in gene expression that signifies both a distinct progenitor type and developmental time point that thereby specifies a particular cell fate at that stage of development. The nervous system is an important setting for understanding developmental timing because different cell types are produced in a certain order and the switch from stem cells to progenitors requires precise timing and regulation. Notable examples of such regulatory molecules include the RNA-binding protein LIN28, and its downstream target, miRNA let-7. Although LIN28 is known to regulate both cell fate and tissue growth, and at times to promote an undifferentiated state, thus far a unified understanding of LIN28’s biological role at the cellular level has not been attained. Here I address LIN28’s activity in mammalian postnatal neurogenesis. Constitutive expression of LIN28 in cells derived from the subventricular zone of the mouse caused several distinct effects: (1) the number of differentiated neurons was dramatically reduced while the relative abundance of two neuronal sub-types was significantly altered; (2) the population of proliferating neural progenitors in the SVZ was reduced while the proportion of neuroblasts was increased, (3) neuro-blast exit from the SVZ increased, and (4) the number of astrocytes was reduced while occasionally causing them to appear early. Thus, LIN28 acts at a post-stem cell/pre-differentiation step, and its continuous expression caused a precocious, not a reiterative phenotype, as is seen in other experimental systems. I made use of a circular RNA sponge that effectively inhibits let-7 activity to address the degree to which LIN28’s effects are due to its inhibition of let-7. Moreover, since LIN41 contributes to a subset of LIN28’s function in C. elegans, I explored whether LIN41 played a role in mammalian neurogenesis. I found that although LIN28 has a multifaceted role in the number and types of cells produced during postnatal neurogenesis, it appears that its action through let-7 accounts for only a fraction of these effects.

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