Research
Research Assistant
Dr. Michael Robinson's Lab
October 2022-December 2024
As an Undergraduate Research Assistant at Miami University, I have gained valuable experience in various laboratory techniques, including lens explants and immunohistochemistry. Working in the lab of Dr. Michael Robinson has provided me with the opportunity to learn from him and his team of graduate students. This experience has forced me to work to comprehend extensive and advanced topics far above my actual class level, requiring me to learn new ways to take in and grasp these difficult concepts. I am also certified in research ethics, compliance, and safety training through the CITI program.
We are currently researching:
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the role of FGF receptors in lens development
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the regulation of the lens transcriptome and chromatin architecture by the FOXE3 gene
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the regulation of lens development of mRNAs
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and the characterization of Zbtb8b knockout mice and cryopreserve Pax6 mutant alleles
Doctoral Undergraduate Opportunity Scholarship (DUOS)
Miami University’s DUOS program is a remarkable opportunity for students to bridge the gap between graduate and undergraduate research. As a participant, I worked collaboratively with a graduate student and Dr. Robinson throughout the spring semester on my developed research proposal. This provided me the opportunity to deepen my understanding of the research process and contribute to our team’s important work. I also had the opportunity to develop new skills and drive meaningful results in a scientific publication of our findings in the research journal Cells.
Exploring Mouse Lens Development: Protein and Gene Analysis with Lens Epithelial Explants
My project delved into the critical biological processes of differentiation, apoptosis, and proliferation, which played pivotal roles in the development of all mammals. These processes were essential for the proper growth, function, and shape of organisms. The lens of the mammalian eye served as a specialized model for understanding differentiation, particularly the transition from monolayered epithelial cells to lens fiber cells, which was crucial for maintaining healthy vision. Disruptions in this differentiation process could lead to eye conditions such as cataracts, anterior segment dysgenesis, and microphthalmia.
The proposal highlighted the use of vitreous and culture media to enable the in vitro study of lens epithelial cells, making it a more accessible and cost-effective research approach. It also outlined an experiment that aimed to investigate the effects of culture time and media type on proliferation, differentiation, and apoptosis in lens explants. Furthermore, it sought to evaluate the resemblance of in vitro lens fiber cell differentiation to in vivo developmental stages, using RNAseq data analysis.
This research had the potential to deepen our understanding of cellular dynamics in the lens and its relevance to mammalian eye development, contributing to advancements in eye health and vision care.
07/19/24 - Research article was published in Cells.
Title: A Transcriptomics Analysis of the Regulation of Lens Fiber Cell
Differentiation in the Absence of FGFRs and PTEN
Authors: Anil Upreti, Stephanie L Padula, Jacob M Weaver, Brad D Wagner,
Allison M Kneller, Anthony L Petulla, Salil A Lachke, Michael L. Robinson *
Figure 1. Complete process. First, remove eyes from P8 FVB/N mice and use the lenses to perform lens epithelial explants in culture media for one day, and then split into both culture media and differentiation media. Allow the explants to culture in the CM and DM for varying lengths of time (D0, D1, D5, D10). Perform immunology and DAPI staining on explants (TUNEL, Ki67, or γ-crystalline, aquaporin, and F-actin) and take the numerical mean of the cells in each category. Reference these values to the RNA sequencing data.
Figure 2. A. The portion of the explant process depicts the removal of the lens from the eye itself (Belecky-Adams, Adler, and Beebe 2002). B. The portion of the explant process depicts the monolayer of epithelial cells being pinned to the dish (West-Mays, Pino, and Lovicu 2010).
Undergraduate Research Forum Poster
