OMB No. 0925-0001/0002 (Rev.

08/12 Approved Through 8/31/2015)

BIOGRAPHICAL SKETCH
Provide the following information for the Senior/key personnel and other significant contributors.
Follow this format for each person. DO NOT EXCEED FIVE PAGES.

NAME: GONG, LIANG-WEI

eRA COMMONS USER NAME (credential, e.g., agency login): lwgong

POSITION TITLE: Associate professor

EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing,
include postdoctoral training and residency training if applicable. Add/delete rows as necessary.)
DEGREE Completion
(if Date FIELD OF STUDY
INSTITUTION AND LOCATION
applicable) MM/YYYY

Nanchang University in China B.Sc. 1990 – 1994 Biology

The 4th Military Medical University in China M.S. 1994 – 1997 Neuroscience
Southern Medical University in China Ph.D. 1997 – 2000 Neuroscience
Cornell University Postdoc 2001 – 2003 Biophysics
Yale University Postdoc 2003 – 2008 Cell Biology

A. Personal statement.
Proposal Goal. The goal of this proposal is to identify whether Ca2+ influx via TRPC5 is critical for synaptic
vesicle endocytosis. The experiments in this proposal are expected to provide the first piece of evidence
on the cellular function of TRPC5 in mature neurons after development. Additionally, by studying the
cooperative roles of TRPC5 and TRPM7 in synaptic vesicle endocytosis, the results from this proposal will
provide a definitive answer to the question as to Ca2+ influx routes of synaptic vesicle endocytosis in
mammalian systems.
Relevant Experiences. I have a broad background in biophysics, cell biology and neurobiology, with
specific training and expertise in the key research areas of this application. As a postdoctoral fellow in Dr.
Manfred Lindau’s lab at Cornell University, I had extensive training in biophysics to explore the regulatory
mechanisms for exocytosis. In Dr. Pietro De Camilli’s lab at Yale Cell Biology, I gained expertise in cell
biology and neurobiology while investigating mechanisms for vesicle endocytosis. As an independent PI, I
have established a unique and specific detection of single vesicle endocytosis, allowing us to distinguish
the molecular mechanisms for the kinetics and the number of single endocytic events. My recently
published work found the kinetics of individual endocytic events shows a Ca2+ dependence that requires
Syt1.
In the last 3 years, we have established detection of synaptic vesicle endocytosis using pHluorin-based
live-cell imaging assays. My lab has also set up recombinant DNA cloning and lentiviral packaging
techniques, which are essentially for us to generate and deliver multiple TRPM7 mutants to chromaffin
cells and neurons in primary cultures. Setting up these molecular biology techniques and live-cell imaging
assays in the lab has been a bit slow for us as a lab with electrophysiological expertise. However, our
long-term research will benefit from these techniques, which allows us to study roles of TRPM7 channels
in endocytosis using a combination of biophysical, live-cell imaging, molecular biology and genetic
approaches. I am happy that we have all the techniques that require completing this proposal in the lab.
Leadership Qualification. I have developed several collaborations to enhance my independent research. In
collaboration with Dr. Wayne Sossin at McGill University in Canada, we have identified the importance of
an interaction between synaptotagmin 1 and dynamin 1 in endocytosis. Currently, I am investigating
potential phosphorylation modulations of TRPM7 in neurons during stimulation, which is in collaboration
with Dr. Stephanie Cologna in Department of Chemistry, an expert in mass spectrometry. I am also
 collaborating with Dr. Wonhwa Cho in Department of Chemistry to understand phosphoinositides in
endocytosis in both neurons and non-neuronal cells.

Positions and Employment and Honors.

2001 - 2003 Postdoctoral Associate, Cornell University (Dr. Manfred Lindau)
2003 - 2008 Postdoctoral Associate, Yale University (Dr. Pietro De Camilli)
2008 - 2015 Assistant Professor in the Department of Biological Sciences, University of Illinois at Chicago
2015 - Associate Professor in the Department of Biological Sciences, University of Illinois at Chicago
Professional Memberships
2003 - Member, Biophysical Society
2002 - Member, Society of Neuroscience

B. Contribution to Science

1) As an independent PI, I have established a unique method to detect clathrin-mediated endocytosis (CME)
of single vesicles in neuroendocrine chromaffin cells, which allows us to dissect out molecular regulations
of the kinetics and the number of unitary CME events, two factors that control the macroscopic rate of
synaptic vesicle endocytosis. My lab has demonstrated that the dynamics of vesicle fission, the last and
arguably most critical step in CME, is Ca2+ dependent but becomes Ca2+ independent in synaptotagmin 1
(Syt1) knock-out cells, indicating the endocytic kinetics of single CME events is regulated through Ca2+ and
Syt1 dependence. In collaboration with Dr. Wayne Sossin in McGill Univerisity, we have shown for the first
time that an interaction between the juxtamembrane region of Syt1 and GTPase dynamin may also be
critical for vesicle fission of CME. Our study supports a model in which actin polymerization may promote
vesicle fission during CME by inducing cholesterol-dependent membrane reorganization, which argues
against the force-generating role of actin polymerization in vesicle fission.
a. Yao LH, Rao Y, Varga K, Wang CY, Xiao P, Lindau M, Gong LW. Synaptotagmin 1 is necessary for the
Ca2+ dependence of clathrin-mediated endocytosis. J Neurosci; 2012; 32:3778-85. PMC23499702
b. Yao LH, Rao Y, Bang C, Kurilova S, Varga K, Wang CY, Weller BD, Cho W, Cheng J, Gong LW. Actin
polymerization does not provide direct mechanical forces for vesicle fission during clathrin-mediated
endocytosis. J Neurosci. 2013; 33(40):15793-8. PMC24089486
c. Varga KT, Jiang Z, Gong LW. Methods for cell-attached capacitance measurements in mouse adrenal
chromaffin cell. J Vis Exp. 2014; 92:e52024. PMC25408421
d. McAdam RL, Varga KT, Jiang Z, Young FB, Blandford V, McPherson PS, Gong LW, Sossin WS. The
juxtamembrane region of synaptotagmin 1 interacts with dynamin 1 and regulates vesicle fission during
compensatory endocytosis in endocrine cells. J Cell Sci. 2015 May 11. pii: jcs.161505. PMC25964652
e. Liu SL, Wang ZG, Hu Y, Xin Y, Singaram I, Gorai S, Zhou X, Shim Y, Min JH, Gong LW, Hay N,
Zhang J, Cho W. Quantitative lipid imaging reveals a new signaling function of
phosphatidylinositol-3,4-bisphophate: isoform- and site-specific activation of Akt. Mol Cell.
2018; 71: 1092-1104. PMC30174291
f. Jiang ZJ, Delaney TL, Zanin MP, Haberberger RV, Pitson SM, Huang J, Alford S, Cologna SM,
Keating DJ, Gong LW. Extracellular and intracellular sphingosine-1-phosphate distinctly
regulate exocytosis in chromaffin cells. J Neurochem 2019 in press.
g. Jiang ZJ, Li WP, Yao, LH, Delaney TL, Grewe BS, McGinley A, Varga K, Alford S, Gong LW.
TRPM7 channel regulates synaptic vesicle endocytosis and couples endocytosis to membrane
depolarization. PLOS Biology (in revision).

2) As a postdoc in Dr. Pietro De Camilli’s lab, I investigated the functions of phosphoinositide metabolism in
vesicle recycling, and I set up the first electro-physiological rig in Dr. De Camilli’s lab to study vesicle
recycling with patch clamping techniques. More specifically, I studied both a kinase and a phosphatase that
regulate PI(4,5)P2 level within the cells: PI(4)P5-kinase type Iγ (PIPK Iγ), and synaptojanin 1. I showed that
PIPK Iγ is crucial for vesicle priming and fusion pore expansion, and that synaptojanin 1 is required for both
constitutive and triggered endocytosis of AMPA receptors postsynaptically. In a close collaboration with Dr.
Derek Toomre at Yale Cell Biology, I also learned live-cell imaging techniques to track protein trafficking
within cells. Also I have applied live-cell imaging methods to examine AMPA receptor endocytosis in
neurons in culture. The experience in live-cell imaging with Dr. Toomre is important for me to set up the
 live-cell imaging method to examine synaptic vesicle endocytosis using pHluorin in neurons in culture in my
own lab.

3) As a postdoc in Dr. Manfred Lindau’s lab (Cornell University), I studied the molecular mechanisms of
exocytosis in chromaffin cells using patch amperometry that allows simultaneous detection of membrane
capacitance and catecholamine release. My studies demonstrated that: (i) the membrane area of a vesicle
is regulated in tandem with its quantal size to keep vesicular neurotransmitter concentration constant; and
(ii) exocytic release of charged transmitters is governed by influx of extracellular Na+ through the fusion
pore, rather than compensated by ion flux through ion channels on vesicle membrane, as is widely believed.
a. Mosharov E, Gong LW, Khanna B, Sulzer D, Lindau M. Intracellular patch electrochemistry, a
technique to measure intracellular metabolites in single cells: regulation of cytosolic catecholamine in
chromaffin cells. J Neurosci 2003; 23: 5834-5845. PMC12843288
b. Dernick G, Gong LW, Tabares L, Alvarez de Toledo G, Lindau M. Patch amperometry: high-resolution
measurements of single-vesicle fusion and release. Nat Methods 2005; 2: 699-708. PMC16118641
c. Gong LW, Hafez I, Alvarez de Toledo G, and Lindau M. Secretory vesicles membrane area is
regulated in tandem with quantal size in chromaffin cells. J Neurosci 2003; 23: 7917-7921.
PMC12944522
d. Gong LW, Alvarez de Toledo G, Lindau M. Exocytotic catecholamine release is not associated with
cation flux through channels in the vesicle membrane but Na+ influx through the fusion pore. Nat Cell
Biol 2007; 9: 915-922. PMC17643118
a. Gong LW, Di Paolo G, Diaz E, Cestra G, Diaz ME, Lindau M, De Camilli P, Toomre D.
Phosphatidylinositol phosphate kinase type I gamma regulates dynamics of large dense-core vesicle
fusion. Proc Natl Acad Sci USA 2005; 102: 5204-5209. PMC15793002
b. Ferguson SM, Brasnjo G, Hayashi M, Wolfel M, Collesi C, Giovedi S, Raimondi A, Gong LW, Paradise
S, O’Toole E, Flavell R, Cremona O, Misesenbock G, Ryan TA, De Camilli P. Synaptic vesicle
recycling requires dynamin 1 during intense synaptic activity. Science 2007; 316: 570-574.
PMC17463283
c. Gong LW & De Camilli P. Regulation of postsynaptic AMPA responses by synaptojanin 1. Proc Natl
Acad Sci USA 2008; 105: 17561-17566. PMC18987319
d. Volpicelli-Daley LA, Lucast L, Gong LW, Liu L, Sasaki J, Sasaki T, Abrams CS, Kanaho Y, De Camilli
P. Phosphatidylinositol-4-phosphate 5-kinases and phosphatidylinositol 4,5-bisphosphate synthesis in
the brain. J Biol Chem 2010; 285: 28708-28714. PMC20622009

4) My early career focused on analgesia, molecular regulations and potential pathological roles of ion
channels in ischemia with their significance in ion channel modulation, analgesia and stroke. For my
Masters degree, I worked on the mechanisms of morphine in analgesia using immunohistochemistry and
electron microscopy. In pursuit of my PhD, I continued in neurobiology, focusing on the regulation of K+
channels and their function in ischemia. Using patch clamp techniques, my work showed that ischemia
induces persistent hyperactivity of large conductance Ca2+-activated potassium channels via oxidation
modulation in CA1 pyramidal neurons.
a. Gong LW, Ding YQ, Wang D, Zheng HX, Qin BZ, Li JS, Kaneko T, Mizuno N. GABAergic synapses on
mu-opioid receptor-expressing neurons in the superficial dorsal horn: an electron microscope study in
the cat spinal cord. Neurosci Lett 1997; 227:33-36. PMC9178852
b. Gong LW, Gao TM, Huang H and Tong ZQ. Redox modulation of large-conductance calcium-
activated potassium channels in pyramidal neurons from adult rat hippocampal CA1 region, Neurosci
Lett 2000; 286: 191-194. PMC10832017
c. Gong LW, Gao TM, Li X, Huang H and Tong ZQ. Augmentation in activities of large-conductance
calcium-activated potassium channels in rat hippocampal CA1 pyramidal neurons after transient
forebrain ischemia. Brain Res 2000; 884: 147-154. PMC11082496
d. Gong LW, Gao TM, Huang H and Tong ZQ. Transient forebrain ischemia induces persistent
hyperactivity of large conductance calcium-activated K+ channels via oxidation modulation in rat
hippocampal CA1 pyramidal neurons, Eur J Neurosci 2002; 15:779-783. PMC11886457

Complete List of Published Work:

http://www.ncbi.nlm.nih.gov/pubmed?term=gong+lw&cmd=DetailsSearch&log$=activity
C. Research Support.

Pending support:
NIH R01NS110533: Gong (PI) 07/01/2019-06/30/2024 ($1,965,165 requested)

Role of vesicular TRP channels in synaptic vesicle endocytosis

(Just submitted the pre-reward JIT to NIH for this proposal and will likely be funded. This proposal has a 13%tile
ranking, which is within the published 16%tile payline of the NINDS for 2019, the primary institute of this proposal)

ONGOING SUPPORT:
NIH R56MH107387 Gong (PI) 09/01/2016-08/31/2017 ($39,3000 total)
-- 1 year of no-cost extension
2+
The role of a Ca -permeable TRP channel in endocytosis
This proposal is examining the physiological role of TRPM7 channels in endocytosis using neuroendocrine cells and
neurons from TRPM7 KO animals.

COMPLETED SUPPORT:
NSF 1145581 Gong (PI) 03/01/2012 – 02/28/2016 ($600,000 total)
National Science Foundation
Studies in clathrin-mediated endocytosis with a millisecond time resolution
This study is examining the role of actin polymerization and cholesterol in vesicle fission during clathrin-mediated
endocytosis, and the role of dynamin 1 in kiss-and-run exocytosis.

BRF SG 2010-06 Gong (PI) 05/01/2010- 04/30/2011 ($40,000 total)

The Brain Research Foundation
The roles of dynamin and actin polymerization in endocytic vesicle biogenesis
This study, using mouse chromaffin cells, examined the role in endocytosis of dynamin GTPase activity and its
inhibition by dynasore and the role of actin polymerization and its disruption by latrunculin B.