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Summary Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b)., Graphical Abstract, Highlights • Generation of cells with uniparental chromosome disomy (UPD) using MADM technology • Isolation of cells with MADM-induced UPD for single-cell downstream applications • Analysis of cortical cells with MADM-induced UPD using scRNA-seq • Analysis of neurons and astrocytes with MADM-induced UPD to probe genomic imprinting, Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments.

The unfolded protein response (UPR) is important for normal cellular homeostasis, development, and physiology. Endoplasmic reticulum stress and the UPR have also been implicated in a variety of diseases and pathologies. Thus the UPR is being studied in many model systems. We describe in this chapter methods for studying the UPR in a mouse model, specifically the cerebral cortex of mouse brain.

Publisher Summary Systematic studies of the developing brain have been made in many species of vertebrates but not on that of the mouse. This chapter is concerned with the part of the multidisciplinary study in which the development of the mouse brain was examined in terms of behavioral observations, alterations of the electrocorticogram, and neurohistological changes of the cerebral cortex. This chapter shows that histological maturation of the mouse cerebral cortex parallels developmental changes in its electrocorticogram and behavior. While Nissl substance is almost invariably present in cortical neurons at birth, they are otherwise quite immature. Only 4 relatively undifferentiated layers may be distinguished in the cortex of the newborn animal. A 5th layer is formed, at 6 days of age, by the separation of small and medium sized pyramidal cells. By this time, also, there is considerable branching of the dendrites of all types of pyramidal cells, and horizontal cells have appeared in the molecular layer. By 15 to 17 days of age, when the ECoG and behavior appear to be mature, the cortical differentiation is likewise complete.