Single-nucleus transcriptome sequencing reveals the identity of neurons that respond to the panic attack.

In nature, moderate innate defensive behaviors (e.g. escaping and freezing) are commonly triggered by imminent threats in human and animal, accompanied with series of physiological responses related with activation of autonomic nervous system (e.g. tachycardia，increased blood pressure, tachypnea). According to many prior researches, posterior hypothalamic nucleus (PHN) with highly cell diversity is recognized to be connected tightly with panic attack. However, the neural substrate of sub-clusters of this important hypothalamic nucleus in panic attack-related defensive behavior has not yet elucidated. In order to justify the hypothesis, we adopted snDNA-seq to screen out a main population, Cbln2+ neurons, manipulating panic attack behaviors in PHN. Subsequently, we illustrated the neural coding mechanism of Cbln2+ PHN neurons for threats stimulus, and indicated the neurobiological functions of neural network of PHN neurons in defensive behaviors. In conclusion, our data suggested that Cbln2+ PHN neurons is sufficient and necessary to regulate panic attack related defensive behaviors in mice. FosTRAP2+/-::Ai9 system was used to label the neurons responded to panic attack specifically. The mice were hit by ‘battle robot’ 30 min continuously, providing controlled, quantitative and multisensory threat stimuli, to induce panic attack related defensive behaviors right after intraperitoneal injection of 50 mg/kg 4-OH Tamoxifen. snRNA-Seq was performed 2 weeks later.