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Description of Two Groups of Dopaminergic Cells Isolated from the Mouse Olfactory Bulb

Angela Pignatelli
Primo
;Fabio Casciano;Nicoletta Bianchi;Simona Capsoni
2023
2023
adult neurogenesis a significant portion are dopaminergic (DA) interneurons located in the glomerular layer (GL). By using transgenic mice expressing eGFP under the control of thetyrosine hydroxylase (TH) promoter, we were able to identify TH-eGFP + cells that were found not only in the GL layer but also in the mitral and external plexiform layer. We also observeda different fluorescence intensity of the TH-eGFP + cells in the two layers. We supposed that these cells could be adult-generated neurons committed to becoming DA but not yet fullydifferentiated. The different levels of fluorescence could possibly reflect varying degrees of maturation of TH-eGFP + cells. Our hypothesis comes from the electrophysiological analysis ofDA neurons located in the glomerular layer (GL) that display full degree of neuronal maturation, whereas eGFP + cells in the mitral layer (ML) exhibit characteristics of immature cells. Toinvestigate whether this difference in maturation could be confirmed at the gene expression level, we used a fluorescence-activated cell sorting technique (FACS) on enzymaticallydissociated cells from the OB. We split the cells into two groups based on their fluorescence level, possibly corresponding to immature ML cells and fully mature DA neurons from the GL.Semiquantitative real-time PCR was performed to detect the expression level of genes associated with the degree of maturation of DA neurons. We demonstrated that cells expressinglow eGFP fluorescence are indeed immature neurons. Our method can be not only utilized to explore the differences between these two groups of DA neurons but also to engineer newneurons for therapeutic approaches such as transplantation into specific areas of the brain affected by Parkinson's disease.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/2613370
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