Microglia,
the
brain's
primary
resident
immune
cell,
exists
in
various
phenotypic
states
depending
on
intrinsic
and
extrinsic
signaling.
Distinguishing
between
these
phenotypes
can
offer
valuable
biological
insights
into
neurodevelopmental
neurodegenerative
processes.
Recent
advances
single-cell
transcriptomic
profiling
have
allowed
for
increased
granularity
better
separation
of
distinct
microglial
states.
While
techniques
such
as
immunofluorescence
RNA
sequencing
(scRNA-seq)
are
available
to
differentiate
functions,
methods
present
notable
limitations,
including
challenging
quantification
methods,
high
cost,
advanced
analytical
techniques.
This
protocol
addresses
limitations
by
presenting
an
optimized
cell
preparation
procedure
that
prevents
ex
vivo
activation
a
flow
cytometry
panel
distinguish
four
from
murine
brain
tissue.
Following
preparation,
fluorescent
antibodies
were
applied
label
1)
homeostatic,
2)
disease-associated
(DAM),
3)
interferon
response
(IRM),
4)
lipid-droplet
accumulating
(LDAM)
microglia,
based
gene
markers
identified
previous
scRNA-Seq
studies.
Stained
cells
analyzed
assess
distribution
function
age
sex.
A
key
advantage
this
is
its
adaptability,
allowing
provided
be
enhanced
using
additional
with
appropriate
analyzer
(i.e.,
Cytek
Aurora
5
laser
spectral
cytometer)
interrogating
different
regions
or
disease
models.
Additionally,
does
not
require
sorting,
resulting
relatively
quick
straightforward
experiment.
Ultimately,
compare
experimental
groups,
state
age,
lower
cost
higher
throughput
than
scRNA-seq.
Key
features
•
Analysis
without
need
imaging,
(LDAM),
(IRM)
microglia
any
region
and/or
model
interest.
modified
incorporate
interest
dyes
when
capable
multiple
color
detections.
Journal of Neuroinflammation,
Год журнала:
2024,
Номер
21(1)
Опубликована: Июнь 5, 2024
Abstract
Microglia,
the
brain’s
resident
macrophages,
maintain
brain
homeostasis
and
respond
to
injury
infection.
During
aging
they
undergo
functional
changes,
but
underlying
mechanisms
their
contributions
neuroprotection
versus
neurodegeneration
are
unclear.
Previous
studies
suggested
that
microglia
sex
dimorphic,
so
we
compared
microglial
in
mice
of
both
sexes.
RNA-sequencing
hippocampal
revealed
more
aging-associated
changes
female
than
male
microglia,
differences
old
young
microglia.
Pathway
analyses
subsequent
validation
assays
a
stronger
AKT-mTOR-HIF1α-driven
shift
glycolysis
among
indicated
C3a
production
detection
was
elevated
especially
females.
Recombinant
induced
AKT-mTOR-HIF1α
signaling
increased
glycolytic
phagocytic
activity
Single
cell
attributed
dimorphism
abundant
disease-associated
(DAM)
mice,
evaluation
an
Alzheimer’s
Disease
mouse
model
metabolic
complement
also
apparent
context
neurodegenerative
disease
strongest
neuroprotective
DAM2
subset.
Collectively,
our
data
implicate
autocrine
C3a-C3aR
reprogramming
DAM
during
aging,
females,
Disease.
Neuron,
Год журнала:
2024,
Номер
112(23), С. 3877 - 3896.e8
Опубликована: Сен. 30, 2024
The
strongest
risk
factors
for
late-onset
sporadic
Alzheimer's
disease
(AD)
include
the
ε4
allele
of
apolipoprotein
E
(APOE),
R47H
variant
triggering
receptor
expressed
on
myeloid
cells
2
(TREM2),
and
female
sex.
Here,
we
combine
APOE4
TREM2
Journal of Neuroinflammation,
Год журнала:
2025,
Номер
22(1)
Опубликована: Янв. 24, 2025
The
brain
presents
various
structural
and
functional
sex
differences,
for
which
multiple
factors
are
attributed:
genetic,
epigenetic,
metabolic,
hormonal.
While
biological
is
determined
by
both
chromosomes
hormones,
little
known
about
how
these
two
interact
to
establish
this
dimorphism.
Sex
differences
in
the
also
affect
its
resident
immune
cells,
microglia,
actively
survey
parenchyma
with
hormones
throughout
life.
However,
microglial
density
distribution,
morphology
ultrastructural
patterns
physiological
conditions
during
adulthood
largely
unknown.
Here,
we
investigated
aforementioned
properties
of
microglia
using
Four
Core
Genotypes
(FCG)
model,
allows
an
independent
assessment
gonadal
chromosomal
effects
four
conditions:
FCG
XX
Tg
XY−
(both
ovaries);
XXSry
XYSry
testes).
We
compared
results
XY
wild-type
(WT)
mice.
In
adult
mice,
focused
our
investigation
on
ventral
hippocampus
across
different
layers:
CA1
stratum
radiatum
(Rad)
lacunosum-moleculare
(LMol),
as
well
dentate
gyrus
polymorphic
layer
(PoDG).
Double
immunostaining
Iba1
TMEM119
revealed
that
influenced
hormones.
show
Rad
LMol
denser
however,
were
densest
WT
PoDG,
ovarian
animals
had
increased
testes
animals.
Additionally,
was
modulated
a
complex
interaction
between
chromosomes,
affecting
their
cellular
soma
arborization
hippocampal
layers.
Moreover,
analysis
showed
make
overall
more
contacts
pre-
post-synaptic
elements
than
Lastly,
markers
stress,
including
mitochondrion
elongation,
dilation
endoplasmic
reticulum
Golgi
apparatus,
mostly
chromosomally
driven.
Overall,
characterized
aspects
normal
found
be
shaped
shading
light
onto
immunity
at
steady-state.
Immunological Reviews,
Год журнала:
2024,
Номер
327(1), С. 8 - 32
Опубликована: Окт. 1, 2024
Neuroinflammation,
characterized
by
a
complex
interplay
among
innate
and
adaptive
immune
responses
within
the
central
nervous
system
(CNS),
is
crucial
in
responding
to
infections,
injuries,
disease
pathologies.
However,
dysregulation
of
neuroinflammatory
response
could
significantly
affect
neurons
terms
function
structure,
leading
profound
health
implications.
Although
tremendous
progress
has
been
made
understanding
relationship
between
processes
alterations
neuronal
integrity,
specific
implications
concerning
both
structure
have
not
extensively
covered,
with
exception
perspectives
on
glial
activation
neurodegeneration.
Thus,
this
review
aims
provide
comprehensive
overview
multifaceted
interactions
key
inflammatory
players,
exploring
mechanisms
through
which
inflammation
influences
functionality
structural
integrity
CNS.
Further,
it
will
discuss
how
these
lead
impairment
functions
architecture
highlight
consequences
caused
dysregulated
functions,
such
as
cognitive
dysfunction
mood
disorders.
By
integrating
insights
from
recent
research
findings,
enhance
our
landscape
set
stage
for
future
interventions
that
transform
current
approaches
preserve
CNS-related
conditions.
Alzheimer’s
disease
(AD)
and
other
age-related
disorders
associated
with
demyelination
exhibit
sex
differences.
In
this
work,
we
used
single-nuclei
transcriptomics
to
dissect
the
contributions
of
chromosomes
gonads
in
AD.
a
mouse
model
demyelination,
identified
roles
modifying
microglia
oligodendrocyte
responses
before
after
myelin
loss.
an
AD-related
expressing
APOE4,
XY
heightened
interferon
(IFN)
response
tau-induced
demyelination.
The
X-linked
gene,
Toll-like
receptor
7
(
Tlr7
),
regulated
sex-specific
IFN
myelin.
Deletion
dampened
differences
while
protecting
against
Administering
TLR7
inhibitor
mitigated
motor
impairment
male
mice,
indicating
that
plays
role
male-biased
type
I
Interferon
aging-
Reviews in Medical Virology,
Год журнала:
2024,
Номер
34(4)
Опубликована: Июнь 21, 2024
Abstract
Global
attention
is
riveted
on
neurodegenerative
diseases
due
to
their
unresolved
aetiologies
and
lack
of
efficacious
therapies.
Two
key
factors
implicated
include
mitochondrial
impairment
microglial
ageing.
Several
viral
infections,
including
Herpes
simplex
virus‐1
(HSV‐1),
human
immunodeficiency
virus
(HIV)
Epstein‐Barr
virus,
are
linked
heightened
risk
these
disorders.
Surprisingly,
numerous
studies
indicate
viruses
induce
aforementioned
precipitating
events.
Hepatitis
C
Virus,
HIV,
respiratory
syncytial
HSV‐1,
Japanese
Encephalitis
Zika
Enterovirus
71
specifically
impact
function,
leading
malfunction.
These
vital
organelles
govern
various
cell
activities
and,
under
specific
circumstances,
trigger
This
article
explores
the
role
infections
in
elucidating
pathogenesis
ailments.
Various
instigate
ageing
via
destruction,
causing
senescent
microglia
exhibit
activated
behaviour,
thereby
inducing
neuroinflammation
contributing
neurodegeneration.
