Abstract:
The ventrolateral medulla (VLM) is a crucial region in the brain for visceral and somatic control, serving as a significant source of synaptic input to the spinal cord. Experimental studies have shown that gene expression in individual VLM neurons is predictive of their function. However, the molecular and cellular organization of the VLM has remained uncertain. This study aimed to create a comprehensive dataset of VLM cells using single-cell RNA sequencing in male and female mice. The dataset was enriched with targeted sequencing of spinally-projecting and adrenergic/noradrenergic VLM neurons. Based on differentially expressed genes, the resulting dataset of 114,805 VLM cells identifies 23 subtypes of neurons, excluding those in the inferior olive, and five subtypes of astrocytes. Spinally-projecting neurons were found to be abundant in seven subtypes of neurons, which were validated through in situ hybridization. These subtypes included adrenergic/noradrenergic neurons, serotonergic neurons, and neurons expressing gene markers associated with premotor neurons in the ventromedial medulla. Further analysis of adrenergic/noradrenergic neurons and serotonergic neurons identified nine and six subtypes, respectively, within each class of monoaminergic neurons. Marker genes that identify the neural network responsible for breathing were concentrated in two subtypes of neurons, delineated from each other by markers for excitatory and inhibitory neurons. These datasets are available for public download and for analysis with a user-friendly interface. Collectively, this study provides a fine-scale molecular identification of cells in the VLM, forming the foundation for a better understanding of the VLM\'s role in vital functions and motor control.
摘要:
腹外侧延髓(VLM)是大脑中内脏和躯体控制的关键区域,作为脊髓突触输入的重要来源。实验研究表明,单个VLM神经元中的基因表达可以预测其功能。然而,VLM的分子和细胞组织仍然不确定。这项研究旨在使用单细胞RNA测序在雄性和雌性小鼠中创建VLM细胞的综合数据集。该数据集富含脊髓投射和肾上腺素能/去甲肾上腺素能VLM神经元的靶向测序。基于差异表达的基因,结果114,805个VLM细胞的数据集识别了23个神经元亚型,不包括那些劣质橄榄,和星形胶质细胞的5个亚型。发现脊髓投射神经元在7种神经元亚型中丰富,通过原位杂交进行了验证。这些亚型包括肾上腺素能/去甲肾上腺素能神经元,血清素能神经元,和表达与腹内侧延髓前运动神经元相关的基因标记的神经元。对肾上腺素能/去甲肾上腺素能神经元和5-羟色胺能神经元的进一步分析确定了9种和6种亚型,分别,在每一类单胺能神经元中。识别负责呼吸的神经网络的标记基因集中在2种神经元亚型中,通过兴奋性和抑制性神经元的标记相互区分。这些数据集可用于公开下载,并通过用户友好的界面进行分析。总的来说,这项研究提供了VLM中细胞的精细分子鉴定,为更好地理解VLM在重要功能和运动控制中的作用奠定了基础。重要性陈述腹外侧延髓(VLM)是大脑的解剖学复杂区域,在调节生命功能中起着至关重要的作用,包括自主和呼吸控制,睡眠-觉醒行为,头颅运动功能,和运动。本研究根据VLM细胞类型和神经元亚型的分子和解剖特征对其进行了全面的分类,通过利用单核RNA测序,RNA荧光原位杂交,和轴突束追踪。我们提供了一个包含114,805个单核转录组的数据集,该数据集鉴定并验证了参与自主和运动系统功能的神经元的精确分子特征。这个公开可用的数据集为全面的实验研究提供了新的机会,以剖析重要的体内平衡功能和身体运动的中央组织。