Impact of Malate Dehydrogenase 2 Skin Epithelial Knockout on Basal Keratinocyte Proliferation
Session Number
Project ID: MEDH 12
Advisor(s)
Ayasa Michii
Rui Yi, Northwestern University, Feinberg School of Medicine
Discipline
Medical and Health Sciences
Start Date
17-4-2024 8:55 AM
End Date
17-4-2024 9:10 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 such as 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. Knockout mice expressed a severe impaired phenotype, notably 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 indicate higher levels of proliferation in knockout mice, with future work required concerning application and analysis with other techniques such as single- cell RNA sequencing.
Impact of Malate Dehydrogenase 2 Skin Epithelial Knockout 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 such as 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. Knockout mice expressed a severe impaired phenotype, notably 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 indicate higher levels of proliferation in knockout mice, with future work required concerning application and analysis with other techniques such as single- cell RNA sequencing.