Proneural transcription factors establish molecular cascades to orchestrate neuronal diversity. One such transcription factor, Atonal homolog 1 (Atoh1), gives rise to cerebellar excitatory neurons and over 30 distinct nuclei in the brainstem critical for hearing, breathing, and balance. Although Atoh1 lineage neurons have been qualitatively described, the transcriptional programs that drive their fate decisions and the full extent of their diversity remain unknown. Here, we analyzed single-cell RNA sequencing and ATOH1 DNA binding in Atoh1 lineage neurons of the developing mouse hindbrain. This high-resolution dataset identified markers for specific brainstem nuclei and demonstrated that transcriptionally heterogeneous progenitors require ATOH1 for proper migration. Moreover, we identified a sizable population of proliferating unipolar brush cell progenitors in the mouse Atoh1 lineage, previously described in humans as the origin of one medulloblastoma subtype. Collectively, our data provide insights into the developing mouse hindbrain and markers for functional assessment of understudied neuronal populations.

前神经转录因子建立分子级联反应以协调神经元多样性。一种这样的转录因子，无调性同源物 1 （Atoh1），在脑干中产生小脑兴奋性神经元和 30 多个不同的细胞核，这对听力、呼吸和平衡至关重要。尽管已经对 Atoh1 谱系神经元进行了定性描述，但驱动其命运决定的转录程序及其多样性的全部程度仍然未知。在这里，我们分析了发育中的小鼠后脑 Atoh1 谱系神经元中的单细胞 RNA 测序和 ATOH1 DNA 结合。这个高分辨率数据集确定了特定脑干核的标志物，并证明转录异质性祖细胞需要 ATOH1 才能正确迁移。此外，我们在小鼠 Atoh1 谱系中鉴定出相当多的增殖单极刷状细胞祖细胞，以前在人类中被描述为一种髓母细胞瘤亚型的起源。总的来说，我们的数据为发育中的小鼠后脑和用于研究不足的神经元群的功能评估标志物提供了见解。