AAV-hChR2 viruses were used for optogenetic activation of excitatory neurons, AAV-hM4D(Gi) virus was used for chemogenetic inhibition experiments. AAV-taCasp3 virus was used for ablating M1 L5 excitatory neurons. PRVs were used for retrograde trans-synaptic tracing experiments.

Calcium sensors	AAV-hSyn-GCaMP6f  AAV-DIO-GCaMP6s
Optogenetic	AAV-CaMKIIα-hChR2 (H134R)-mCherry  AAV-DIO-hChR2(H134R)-mCherry
Control	AAV-hSyn-mCherry  AAV-CaMKIIα-EGFP  AAV-DIO-mCherry
Chemogenetics	AAV-CaMKIIa-hM4D(Gi)-mCherry
CRE Recombinase	AAV-CaMKIIa-Cre-GFP  AAV1-hSyn-Cre  AAV1-CMV-Cre-GFP
Neuron Ablation	AAV-CAG-DIO-taCasp3-TEVp
PRV	PRV-EGFP  PRV-RFP

Jiwei Yao, Quanchao Zhang, Xiang Liao, Qianwei Li, Shanshan Liang, Xianping Li, Yalun Zhang, Xiangning Li, Haoyu Wang, Han Qin, Meng Wang, Jingcheng Li, Jianxiong Zhang, Wenjing He, Wen Zhang4, Tong Li, Fuqiang Xu, Hui Gong, Hongbo Jia, Xiaohong Xu, Junan Yan   and Xiaowei Chen
Pub Date: 2018-10-25,  DOI: 10.1038/s41593-018-0256-4，Email: sales@brainvta.com
Urination (also called micturition) is thought to be regulated by a neural network that is distributed in both subcortical and cortical regions. Previously, urination-related neurons have been identified in subcortical structures such as the pontine micturition center (also known as Barrington’s nucleus). However, the origin of the descending cortical pathway and how it interfaces with this subcortical circuit to permit voluntary initiation of urination remain elusive. Here we identified a small cluster of layer 5 neurons in the primary motor cortex whose activities tightly correlate with the onset of urination in freely behaving mice and increase dramatically during territorial marking. Optogenetically activating these neurons elicits contraction of the bladder and initiates urination, through their projections to the pontine micturition center, while silencing or ablating them impairs urination and causes retention of urine. Together these results reveal a novel cortical component upstream of the pontine micturition center that is critically involved in urination.