Selected Publications
Widespread co-release of glutamate and GABA throughout the mouse brain
Cesar C. Ceballos, Lei Ma, Maozhen Qin & Haining Zhong. Communications Biology (2024). https://doi.org/10.1038/s42003-024-07198-y.
"Here, we investigated GABA and glutamate co-release using electrophysiological recordings of miniature postsynaptic currents. We found that biphasic minis involving both AMPA and GABAA receptors were widespread throughout many brain regions.”
Check out paper here.
Locomotion activates PKA through dopamine and adenosine in striatal neurons
Lei Ma, Julian Day-Cooney, Omar Jáidar Benavides, Michael A. Muniak, Maozhen Qin, Jun B. Ding, Tianyi Mao & Haining Zhong. Nature (2022). https://doi.org/10.1038/s41586-022-05407-4
“Our results suggest that acute adenosine accumulation interplays with dopamine release to orchestrate protein kinase A (PKA) activity in striatal spiny projection neurons (SPNs) and proper striatal function during animal locomotion.”
Check out the full article in Nature here.
Sensitive genetically encoded sensors for population and subcellular imaging of cAMP in vivo
Crystian I. Massengill, Landon Bayless-Edwards, Cesar C. Ceballos, Elizabeth R. Cebul, James Cahill, Arpita Bharadwaj, Evan Wilson, Maozhen Qin, Matthew R. Whorton, Isabelle Baconguis, Bing Ye, Tianyi Mao & Haining Zhong (2022) Nat Methods. https://doi.org/10.1038/s41592-022-01646-5
Published in Nature Methods, the Zhong Lab and collaborators have developed a genetically encoded cAMP sensor, cleverly dubbed “cAMPFIRE”. This sensor can detect nanomolar concentrations of cAMP, can be used in vivo, and is compatible with both ratiometric and fluorescence lifetime imaging. The cAMPFIRE sensor will be a valuable tool for the scientific community, and we couldn’t be more excited about it.
Check out the paper here.
Labeling Endogenous Proteins Using CRISPR-mediated Insertion of Exon (CRISPIE)
Wilson, E., Mao, T. and Zhong, H. (2022) Bio-protocol.
This published bio-protocol details the background, materials, and protocol for performing the CRISPIE genome editing methodology developed in Zhong et al. 2021. Check out the protocol here. Bio-protocol is a journal that publishes detailed protocols from previously published papers, with the aim of improving reproducibility and transparency in science.
Genetically encoded sensors towards imaging cAMP and PKA activity in vivo
Crystian I Massengill, Julian Day-Cooney, Tianyi Mao, Haining Zhong (2021). J Neurosci Methods.
This review summarizes genetically encoded cAMP and PKA sensors for in vivo imaging.
Distinct in vivo dynamics of excitatory synapses onto cortical pyramidal neurons and inhibitory interneurons
Melander JB, Nayebi A, Jongbloets BC, Fortin DA, Qin M, Ganguli S, Mao T, Zhong H (2021). Cell Reports.
Here, we fluorescently visualize PSD-95 at endogenous levels in the mouse barrel cortex. Longitudinal in vivo imaging reveals that, while synaptic weights in both pyramidal and PV+ cells are log-normally distributed, synapses onto PV+ neurons are less heterogeneous and more stable. These results reveal that cell type-specific processes govern cortical synaptic strengths and dynamics.
High-fidelity, efficient, and reversible labeling of endogenous proteins using CRISPR-based designer exon insertion
Zhong H, Massengill CI, Muniak MA, Ma L, Qin M, Kaech Petrie S, Mao T. (2020). eLife.
Here, we describe a method called CRISPR-mediated insertion of exon (CRISPIE) that can precisely and reversibly label endogenous proteins using CRISPR/Cas9-based editing.
Myristoylation alone is sufficient for PKA catalytic subunits to fractionally associate with the plasma membrane to regulate neuronal functions
Xiong W-H, Qin M, Zhong H. (2020). PNAS.
Here, we report that the electrically neutral N-terminal fragment of the protein kinase A catalytic subunit (PKA-C), in which myristoylation is the only functional motif, is sufficient for membrane association. This myristoylation can associate a fraction of PKA-C molecules or fluorescent proteins (FPs) to the plasma membrane in neuronal dendrites.
Visualizing Protein Kinase A activity in head-fixed behaving mice using in vivo two-photon fluorescence lifetime imaging microscopy
Jongbloets BC, Ma L, Mao T, Zhong H. (2019) J. Vis. Exp. (148)
A procedure is presented to visualize protein kinase A activities in head-fixed, behaving mice. Two-photon fluorescence lifetime imaging microscopy is used to visualize PKA activities in vivo during enforced locomotion.
A Highly Sensitive A-Kinase Activity Reporter for Imaging Neuromodulatory Events in Awake Mice
Ma L, Jongbloets BC, Xiong WH, Melander JB, Qin M, Lameyer TJ, Harrison MF, Zemelman BV, Mao T*, Zhong H*. (2018). Neuron 99:665-679.e5.
Here, we developed a PKA sensor, tAKARα, which enables detection of physiologically relevant activity and interrogation of neuromodulation-induced PKA-signaling in awake animals.
Ultrafast neuronal imaging of dopamine dynamics with designed genetically encoded sensors
Patriarchi T, Cho JR, Merten K, Howe MW, Marley A, Xiong WH, Folk RW, Broussard GJ, Liang R, Jang MJ, Zhong H, Dombeck D, von Zastrow M, Nimmerjahn A, Gradinaru V, Williams JT, Tian L. (2018). Science 360:eaat4422.
Patriarchi et al. developed fluorescent protein–based dopamine indicators to visualize spatial and temporal release of dopamine directly with high fidelity and resolution. In the cortex, two-photon imaging with these indicators was used to map dopamine activity at cellular resolution.
Liberated PKA catalytic subunits associate with the membrane via myristoylation to preferentially phosphorylate membrane substrates.
Tillo SE, Xiong WH, Takahashi M, Miao S, Andrade AL, Fortin DA, Yang G, Qin M, Smoody BF, Stork PJS, Zhong H. (2017). Cell Reports 19:617-629.
We report that elevation of cyclic AMP in neuronal dendrites causes a significant percentage of the PKA catalytic subunit (PKA-C) molecules to be released from the regulatory subunit (PKA-R). We propose that activation-dependent association of PKA-C renders the membrane a unique PKA-signaling compartment.
A comprehensive excitatory input map of the striatum reveals novel functional organization
Hunnicutt BJ, Jongbloets BC, Birdsong WT, Gertz KJ, Zhong H, Mao T. (2016) eLife 5:e19103.
This study presents a comprehensive map of the excitatory inputs to the mouse striatum. The input patterns reveal boundaries between the known striatal domains The complete thalamo-cortico-striatal loop is also presented.
Applying Superresolution Localization-Based Microscopy to Neurons
Zhong H. (2015). Synapse. 69:283-294.
Here, we discuss the principles of LBM, its current applications in neuroscience, and the challenges that must be met before its full potential is achieved. We also present the unpublished results of our own experiments to establish a sample preparation procedure for applying LBM to study brain tissue.
Live Imaging of Endogenous PSD-95 Using ENABLED: A Conditional Strategy to Fluorescently Label Endogenous Proteins
Fortin DA, Tillo SE, Yang G, Rah JC, Melander JB, Bai S, Soler-Cedeño O, Qin M, Zemelman BV, Guo C, Mao T*, Zhong H*. (2014). J. Neurosci. 34 (50) 16698-16712
Here, we describe a conditional mouse genetic strategy termed endogenous labeling via exon duplication (ENABLED), which can be used to fluorescently label endogenous proteins with near ideal properties in all neurons, a sparse subset of neurons, or specific neuronal subtypes.
A comprehensive thalamocortical projection map at the mesoscopic level
Hunnicutt BJ, Long BR, Kusefoglu D, Gertz KJ, Zhong H*, Mao T*. (2014). Nat. Neurosci. 17:1276-1285.
We employed a systematic, high-throughput viral approach to visualize thalamocortical axons with high sensitivity. We then developed algorithms to directly compare injection and projection information across animals and constructed a comprehensive map of thalamocortical projections.