bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2022,
Volume and Issue:
unknown
Published: Dec. 27, 2022
Abstract
Drosophila
has
been
a
powerful
model
system
for
biological
studies
due
to
the
wide
range
of
genetic
tools
established
it.
Among
these
tools,
Gal4
is
most
abundant,
offering
unparalleled
tissue-
and
developmental
stage-specificity
gene
manipulation.
In
comparison,
other
reagents
are
far
fewer
in
choices.
Here
we
present
toolkit
converting
strains
into
LexA
Flippase
transgenes
through
simple
crosses
fluorescence
screening.
We
demonstrate
proof-of-principle
by
ten
lines
that
exhibit
diverse
tissue
specificities
examined
activity
patterns
converted
lines.
Gal4-to-LexA
Flp
conversion
fast
convenient
should
greatly
expand
choices
binary
expression
FRT-based
mosaic
analysis,
respectively,
.
Proceedings of the National Academy of Sciences,
Journal Year:
2024,
Volume and Issue:
121(16)
Published: April 8, 2024
The
ability
of
neurons
to
rapidly
remodel
their
synaptic
structure
and
strength
in
response
neuronal
activity
is
highly
conserved
across
species
crucial
for
complex
brain
functions.
However,
mechanisms
required
elicit
coordinate
the
acute,
activity-dependent
structural
changes
synapses
are
not
well
understood,
as
neurodevelopment
plasticity
tightly
linked.
Here,
using
an
RNAi
screen
Drosophila
against
genes
affecting
nervous
system
functions
humans,
we
uncouple
cellular
processes
important
synapse
development.
We
find
mutations
associated
with
neurodegenerative
mental
health
disorders
2-times
more
likely
affect
activity-induced
remodeling
than
report
that
while
both
development
at
fly
NMJ
require
macroautophagy
(hereafter
referred
autophagy),
bifurcation
autophagy
pathway
differentially
impacts
plasticity.
demonstrate
enhances
activation
but
diminishes
degradative
autophagy,
thereby
driving
towards
autophagy-based
secretion.
Presynaptic
knockdown
Snap29,
Sec22,
or
Rab8,
proteins
implicated
secretory
pathway,
sufficient
abolish
remodeling.
This
study
uncovers
a
transsynaptic
signaling
mechanism
modulating
Nature Communications,
Journal Year:
2021,
Volume and Issue:
12(1)
Published: April 23, 2021
Abstract
Lipid
shuttling
between
neurons
and
glia
contributes
to
the
development,
function,
stress
responses
of
nervous
system.
To
understand
how
a
neuron
acquires
its
lipid
supply
from
specific
lipoproteins
their
receptors,
we
perform
combined
genetic,
transcriptome,
biochemical
analyses
in
developing
Drosophila
larval
brain.
Here
report,
astrocyte-derived
secreted
lipocalin
Glial
Lazarillo
(GLaz),
homolog
human
Apolipoprotein
D
(APOD),
neuronal
receptor,
brain-specific
short
isoforms
lipophorin
receptor
1
(LpR1-short),
cooperatively
mediate
neuron-glia
support
dendrite
morphogenesis.
The
isoform
specificity
LpR1
defines
distribution,
binding
partners,
ability
proper
growth
synaptic
connectivity.
By
demonstrating
physical
functional
interactions
GLaz/APOD
LpR1,
elucidate
molecular
pathways
mediating
trafficking
fly
brain,
provide
vivo
evidence
indicating
isoform-specific
expression
lipoprotein
receptors
as
key
mechanism
for
regulating
cell-type
recruitment.
Abstract
Drosophila
has
been
a
powerful
model
system
for
biological
studies
due
to
the
wide
range
of
genetic
tools
established
it.
Among
these
tools,
Gal4
is
most
abundant,
offering
unparalleled
tissue
and
developmental
stage
specificity
gene
manipulation.
In
comparison,
other
reagents
are
far
fewer
in
choices.
Here
we
present
toolkit
converting
strains
into
LexA
Flippase
transgenes
through
simple
crosses
fluorescence
screening.
We
demonstrate
proof-of-principle
by
ten
lines
that
exhibit
diverse
specificities
examined
activity
patterns
converted
lines.
Gal4-to-LexA
Flp
conversion
fast
convenient
should
greatly
expand
choices
binary
expression
FRT-based
mosaic
analysis,
respectively,
Drosophila.
Science Advances,
Journal Year:
2025,
Volume and Issue:
11(11)
Published: March 12, 2025
Lipid
homeostasis
is
critical
to
neuronal
survival.
ATP-binding
cassette
A
(ABCA)
proteins
are
lipid
transporters
associated
with
neurodegenerative
diseases.
How
ABCA
regulate
in
neurodegeneration
an
outstanding
question.
Here
we
report
that
the
Drosophila
protein
engulfment
ABC
transporter
ovary
(Eato)
regulates
phagocytosis-dependent
by
playing
opposing
roles
neurons
and
phagocytes:
In
neurons,
Eato
prevents
dendrites
axons
from
being
attacked
neighboring
phagocytes;
phagocytes,
sensitizes
cell
for
detecting
as
targets.
Thus,
deficiency
alone
causes
neurite
degeneration,
but
additional
loss
phagocytes
suppresses
degeneration.
Mechanistically,
functions
removing
eat-me
signal
phosphatidylserine
surface
both
phagocytes.
Multiple
human
worm
homologs
can
rescue
not
suggesting
conserved
type–specific
activities
of
proteins.
These
results
imply
possible
mechanisms
neuron-phagocyte
interactions
Learning & Memory,
Journal Year:
2024,
Volume and Issue:
31(5), P. a053863 - a053863
Published: May 1, 2024
How
does
the
brain
translate
sensory
information
into
complex
behaviors?
With
relatively
small
neuronal
numbers,
readable
behavioral
outputs,
and
an
unparalleled
genetic
toolkit,
Drosophila
mushroom
body
(MB)
offers
excellent
model
to
address
this
question
in
context
of
associative
learning
memory.
Recent
technological
breakthroughs,
such
as
freshly
completed
full-brain
connectome,
multiomics
approaches,
CRISPR-mediated
gene
editing,
machine
techniques,
led
major
advancements
our
understanding
MB
circuit
at
molecular,
structural,
physiological,
functional
levels.
Despite
significant
progress
individual
areas,
field
still
faces
fundamental
challenge
resolving
how
these
different
levels
combine
interact
ultimately
control
behavior
fly.
In
review,
we
discuss
various
aspects
research,
with
a
focus
on
current
knowledge
gaps,
outlook
future
methodological
developments
required
reach
overall
view
neurobiological
basis
Methods in molecular biology,
Journal Year:
2022,
Volume and Issue:
unknown, P. 157 - 176
Published: Jan. 1, 2022
Abstract
Over
the
last
century
research
in
Drosophila
has
resulted
many
fundamental
contributions
to
our
understanding
of
biology
multicellular
organisms.
Many
these
breakthroughs
have
been
based
on
identification
novel
gene
functions
large-scale
genetic
screens.
However,
conventional
forward-genetic
screens
limited
by
random
nature
mutagenesis
and
difficulties
mapping
causal
mutations,
while
reverse-genetic
RNAi
suffer
from
incomplete
knockdown
expression.
Recently
developed
CRISPR-Cas9
libraries
promise
address
limitations
allowing
induction
targeted
mutations
genes
with
spatial
temporal
control.
Here,
we
provide
a
guide
for
tissue-specific
CRISPR
screening
,
including
characterization
Gal4
UAS-Cas9
lines,
selection
sgRNA
libraries,
various
quality
control
measures.
We
also
discuss
confounding
factors
that
can
give
rise
false-positive
false-negative
results
such
experiments
suggest
strategies
how
detect
avoid
them.
Conditional
represents
an
exciting
new
approach
functional
genomics
vivo
is
set
further
expand
knowledge
molecular
underpinning
development,
homeostasis,
disease.
Developmental Cell,
Journal Year:
2024,
Volume and Issue:
59(19), P. 2672 - 2686.e5
Published: July 5, 2024
CRISPR-Cas
greatly
facilitated
the
integration
of
exogenous
sequences
into
specific
loci.
However,
knockin
generation
in
multicellular
animals
remains
challenging,
partially
due
to
complexity
insertion
screening.
Here,
we
describe
SEED/Harvest,
a
method
generate
knockins
Drosophila,
based
on
and
single-strand
annealing
(SSA)
repair
pathway.
In
SEED
(from
"scarless
editing
by
element
deletion"),
switchable
cassette
is
first
integrated
target
locus.
subsequent
CRISPR-triggered
event,
resolved
SSA,
seamlessly
removed.
Germline
excision
cassettes
allows
for
fast
robust
both
fluorescent
proteins
short
protein
tags
tandem.
Tissue-specific
expression
Cas9
results
somatic
excision,
conferring
spatiotemporal
control
labeling
conditional
rescue
mutants.
Finally,
achieve
manipulation
tag
knockins,
developed
genetic
toolbox
functionalizing
ALFA
nanobody.
