The Wang Lab employs multidisciplinary approaches, including but not limited to electrophysiology, real-time imaging, optogenetics, neuronal tracing, single-cell RNA sequencing (RNA-seq), mass spectrometry (mass-spec), and animal behavior analysis, to study hypothalamic neural circuits. Additionally, we have developed novel technologies to facilitate our research:

Anterograde Trans-synaptic Tracing

The brain coordinates animal physiology and behavior through interconnected neurons. To understand the structural basis of signal transduction within neural networks, it is essential to delineate the synaptic connectivity between neurons. Trans-synaptic tracers serve as powerful tools for this purpose. While retrograde tracers are widely used in circuit mapping, there have been limited anterograde tracing tools to complement their retrograde counterparts. Here, we developed an anterograde trans-synaptic tracing toolkit, named WTR, that delivers recombinase from genetically defined starter neurons to their downstream targets. Accompanied by postsynaptic expression of Cre/Flpo-dependent payloads such as GCaMP7s or ChR2, the WTR enables labeling, recording, or manipulation of downstream neurons. This toolkit will facilitate systematic research into the architecture and function of neural circuits.

Single-cell RNA-seq

The heterogeneity of neurons in the brain poses one of the major challenges in studying neuronal functions associated with diverse physiological and behavioral processes. Single-cell RNA sequencing (RNA-seq), since its initial application in neuroscience research, has facilitated neural circuit mapping by providing genetic access to neuronal subpopulation of interest. The Wang Lab optimized the SMART-seq3 protocol and established a high-quality, low-cost pipeline, which yields sequencing libraries of > 10,000 genes per cell (mature neurons), covering full-length transcriptomes. By utilizing automated liquid handlers, this pipeline achieves a throughput suitable for research projects involving thousands of cells. This method has become a powerful tool in neural circuit research and beyond.