bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 23, 2024
Neurochemical
signals
like
dopamine
(DA)
play
a
crucial
role
in
variety
of
brain
functions
through
intricate
interactions
with
other
neuromodulators
and
intracellular
signaling
pathways.
However,
studying
these
complex
networks
has
been
hindered
by
the
challenge
detecting
multiple
neurochemicals
vivo
simultaneously.
To
overcome
this
limitation,
we
developed
single-protein
chemigenetic
DA
sensor,
HaloDA1.0,
which
combines
cpHaloTag-chemical
dye
approach
G
protein-coupled
receptor
activation-based
(GRAB)
strategy,
providing
high
sensitivity
for
DA,
sub-second
response
kinetics,
an
extensive
spectral
range
from
far-red
to
near-infrared.
When
used
together
existing
green
red
fluorescent
neuromodulator
sensors,
Ca2+
indicators,
cAMP
optogenetic
tools,
HaloDA1.0
provides
versatility
multiplex
imaging
cultured
neurons,
slices,
behaving
animals,
facilitating
in-depth
studies
dynamic
neurochemical
networks.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 12, 2025
The
concentrations
of
extracellular
and
intracellular
signaling
molecules,
such
as
dopamine
cAMP,
change
over
both
fast
slow
timescales
impact
downstream
pathways
in
a
cell-type
specific
manner.
Fluorescence
sensors
currently
used
to
monitor
signals
vivo
are
typically
optimized
detect
fast,
relative
changes
concentration
the
target
molecule.
They
less
well
suited
slowly-changing
rarely
provide
absolute
measurements
either
components.
Here,
we
developed
system
for
fluorescence
lifetime
photometry
at
high
temporal
resolution
(FLIPR)
that
utilizes
frequency-domain
analog
processing
measure
genetically-encoded
speed
but
with
long-term
stability
picosecond
precision
freely
moving
mice.
We
applied
FLIPR
investigate
two
functionally
distinct
regions
striatum,
nucleus
accumbens
core
(NAC)
tail
striatum
(TOS).
observed
higher
tonic
levels
baseline
TOS
compared
NAC
detected
differential
dynamic
responses
phasic
appetitive
aversive
stimuli.
Thus,
enables
simple
monitoring
time-scale
neuronal
units,
revealing
previously
unappreciated
spatial
variation
even
well-studied
systems.
Journal of Cell Science,
Journal Year:
2025,
Volume and Issue:
138(1)
Published: Jan. 1, 2025
ABSTRACT
G
protein-coupled
receptor
(GPCR)
signalling
pathways
underlie
numerous
physiological
processes,
are
implicated
in
many
diseases
and
major
targets
for
therapeutics.
There
more
than
800
GPCRs,
which
together
transduce
a
vast
array
of
extracellular
stimuli
into
variety
intracellular
signals
via
heterotrimeric
protein
activation
multiple
downstream
effectors.
A
key
challenge
cell
biology
research
the
pharmaceutical
industry
is
developing
tools
that
enable
quantitative
investigation
GPCR
to
gain
mechanistic
insights
varied
cellular
functions
pharmacology
GPCRs.
Recent
progress
this
area
has
been
rapid
extensive.
In
Review,
we
provide
critical
overview
these
new,
state-of-the-art
approaches
investigate
pathways.
These
include
novel
sensors,
Förster
or
bioluminescence
resonance
energy
transfer
assays,
libraries
tagged
proteins
transcriptional
reporters.
improved
studies
different
stages
signalling,
including
activation,
second
messenger
(cAMP
Ca2+)
β-arrestin
recruitment
internalisation
trafficking
Chemical Reviews,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 25, 2025
Cell-surface
receptors
are
vital
for
controlling
numerous
cellular
processes
with
their
dysregulation
being
linked
to
disease
states.
Therefore,
it
is
necessary
develop
tools
study
and
the
signaling
pathways
they
control.
This
Review
broadly
describes
molecular
approaches
that
enable
1)
visualization
of
determine
localization
distribution;
2)
sensing
receptor
activation
permanent
readouts
as
well
in
real
time;
3)
perturbing
activity
mimicking
receptor-controlled
learn
more
about
these
processes.
Together,
have
provided
valuable
insight
into
fundamental
biology
helped
characterize
therapeutics
target
receptors.
Journal of Cell Science,
Journal Year:
2024,
Volume and Issue:
137(20)
Published: Oct. 15, 2024
In
2023,
the
ImaBio
consortium
(imabio-cnrs.fr),
an
interdisciplinary
life
microscopy
research
group
at
Centre
National
de
la
Recherche
Scientifique,
celebrated
its
20th
anniversary.
contributes
to
biological
imaging
community
through
organization
of
MiFoBio
conferences,
which
are
conferences
featuring
lectures
and
hands-on
workshops
that
attract
specialists
from
around
world.
provide
with
opportunity
reflect
on
evolution
field,
2023
event
offered
retrospective
talks
discussing
past
20
years
topics
in
microscopy,
including
multicellular
assemblies,
image
analysis,
quantification
molecular
motions
interactions
within
cells,
advancements
fluorescent
labels,
laser
technology
for
multiphoton
label-free
thick
samples.
this
Perspective,
we
compile
summaries
these
presentations
overviewing
a
specific
area
each
concludes
brief
look
towards
future.
The
full
available
YouTube
channel
(youtube.com/@gdrimabio5724).
