MYT1L deficiency impairs excitatory neuron trajectory during cortical development [P1]
Mutations that reduce the function of MYT1L, a neuron-specific transcription factor, are associated with a syndromic neurodevelopmental disorder. Furthermore, MYT1L is routinely used as a pro-neural factor in fibroblast-to-neuron transdifferentiation. MYT1L has been hypothesized to play a role in the trajectory of neuronal specification and subtype specific maturation, but this hypothesis has not been directly tested, nor is it clear which neuron types are most impacted by MYT1L loss. In this study, we profiled 412,132 nuclei from the forebrains of wild-type and MYT1L-deficient mice at three developmental stages: E14 at the peak of neurogenesis, P1 when neurons in the six cortical layers have been born, and P21 when neurogenesis is complete and neurons are maturing, to examine the role of MYT1L levels in the trajectory of neuronal development. We found that MYT1L deficiency significantly disrupted the relative proportions of cortical excitatory neurons. Changes in gene expression were concentrated in excitatory neurons, suggesting that transcriptional effects of MYT1L deficiency are largely due to disruption of neuronal maturation programs. Most effects on gene expression were cell autonomous and persistent through development. In addition, while MYT1L can both activate and repress gene expression, the repressive effects were most sensitive to haploinsufficiency, and thus more likely mediate MYT1L syndrome. These findings illuminate the intricate role of MYT1L in orchestrating gene expression dynamics during neuronal development, providing insights into the molecular underpinnings of MYT1L syndrome. Overall design: Samples for snRNAseq were collected at three key developmental timepoints: E14, P1, and P21. At E14, forebrains were dissected from MYT1L WT, Het, and KO mice, with three replicates per genotype. These E14 samples included a mix of males and females for each genotype. For P1 samples, forebrains were harvested from 8 WT mice (5 male, 3 female) and 4 Het mice (3 male, 1 female). At P21, cortices were colleced from 6 WT animals (3 male, 3 female) and 6 Het animals (3 male, 3 female). Across all time points, nuclei were isolated from the dissected tissues. Library preparation was carried out using the Scale Biosciences Single Cell RNA Kit, with samples batched by age.