Impact of the Skin Epithelial Knockout of Malate Dehydrogenase 2 on Basal Keratinocyte Proliferation
Session Number
Project ID: MEDH 21
Advisor(s)
Dr. Rui Yi; Northwestern University, Feinberg School of Medicine
Ayasa Michii; Northwestern University, Feinberg School of Medicine
Discipline
Medical and Health Sciences
Start Date
19-4-2023 10:05 AM
End Date
19-4-2023 10:20 AM
Abstract
As a barrier to the external environment, the skin serves as the body’s primary mechanism for physical and chemical defense, thermoregulation, and fluid retention. In both tissue homeostasis and wound repair, layers of skin form as cells migrate upward following proliferation and differentiation in the basal layer. Metabolism maintains physiological activity, with inhibited function in diseases including diabetes leading to conditions like chronic wound healing. Malate dehydrogenase is the final enzyme in the citric acid cycle that converts malate to oxaloacetate, restarting the cycle. Utilizing Cre-Lox recombination, conditional knockout mice lacking epidermis expression of Malate Dehydrogenase 2 were compared to wild type mice for 30 days, with the knockout phenotype notably having a smaller size, wrinkly skin, and damaged hair formation. Mouse back skin slide samples from P5, P7, P14, P20, and P30 were prepared in OCT compound for immunofluorescence utilizing EdU, Ki-67, and Phospho-histone H3 markers costained with Keratin 5 to investigate how impaired metabolism may affect stages of the cell cycle in the basal layer over time. Initial findings show higher activity in DNA replication and mitosis for knockout mice, with future work concerning quantitative analysis and application to other techniques such as Single-cell RNA sequencing data analysis.
Impact of the Skin Epithelial Knockout of Malate Dehydrogenase 2 on Basal Keratinocyte Proliferation
As a barrier to the external environment, the skin serves as the body’s primary mechanism for physical and chemical defense, thermoregulation, and fluid retention. In both tissue homeostasis and wound repair, layers of skin form as cells migrate upward following proliferation and differentiation in the basal layer. Metabolism maintains physiological activity, with inhibited function in diseases including diabetes leading to conditions like chronic wound healing. Malate dehydrogenase is the final enzyme in the citric acid cycle that converts malate to oxaloacetate, restarting the cycle. Utilizing Cre-Lox recombination, conditional knockout mice lacking epidermis expression of Malate Dehydrogenase 2 were compared to wild type mice for 30 days, with the knockout phenotype notably having a smaller size, wrinkly skin, and damaged hair formation. Mouse back skin slide samples from P5, P7, P14, P20, and P30 were prepared in OCT compound for immunofluorescence utilizing EdU, Ki-67, and Phospho-histone H3 markers costained with Keratin 5 to investigate how impaired metabolism may affect stages of the cell cycle in the basal layer over time. Initial findings show higher activity in DNA replication and mitosis for knockout mice, with future work concerning quantitative analysis and application to other techniques such as Single-cell RNA sequencing data analysis.