Download Identification of Primary Lysosomes in Human

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Identification
of Primary
Lysosomes
and
By Mary
The
presence
well
documented;
of lysosomal
has
been
the
determine
tion
the
subject
the
of
two
time
platelets,
and
blood
we
examined
was
degree
reaction
product
was
Golgi-associated
were
sometimes
reaction
product
was
LOOD
bodies
to
reactive.
to
vesicles
of
a-granules
storage
are
in
with
one
or
other
a
two
enzymes,
phosphate
arylsulfatase
bone
undergoing
From
chest
the
Berkeley,
nia School
in part
mammals
cavity
directly
Dense
drawn
from
10 ml
into
to be lysocytochemical
Address
ment
San
surgery.
Cells
ofMedicine,
in part
April
reprint
of Pathology,
Francisco,
/4,
under
1981;
requests
University
Francisco,
Calif
94143.
by Grune
& Stratton,
0006-9971/5903-0003$O1.OO/O
the
Berkeley
fixed
with
M
fixation,
detail
the
buffer,
incubation
according
preparations
were
(while
suspended
water
bath
postfixed
was
pipetted
at
Cells
to
either
room
washed
7.4,
method
in 0504;
6.0;
10
of
wash
stained
with
dehydrated
in
before
uranyl
buffer,
pH
ethanols;
Sections
citrate,
were cut with a diamond
and examined
with a Siemens
cells
then
30
mm.
After
in 0.1
AS
are
described
in
for trimetaphosphatase
Oliver.’8
without
mm
When
to cytochemical
4#{176}Cfor
and
incubated
incubated
CaCI2.16
into fixative;
at 22#{176}C,and
7% sucrose.
AcPase
were
Karnovsky’s
paraformalde-
( 1 hr to overnight)
containing
for
of
0.025%
The
Na-cacodylate
M
Prior
at
was
centrifuge;
1 .9%
dripped
to cool
out
in the postfixation
90#{176}Cfor
0.1
temperature.
carried
directly
blood
5- 1 0 vol of fixative.
and
with
was
either
U heparin/
the
or a modification
buffer
Blood
into
clinical
in
glutaraldehyde
the
bones.
used,
into
glutaraldehyde
pH
platelet
of anticoagulants
g on a table-top
procedures
elsewhere.’2”7
(TMPase)
use
ACD
the cells were thoroughly
M Na-cacodylate
The
was
in
primary
during
(50-100
then
at that
are
or a heparinized
1% sucrose,
fixation
participated
they
volunteers
Na-cacodylate
for 3 hr more
incubations,
had
on the
were omitted,
blood
was
30 mm at 37#{176}C,allowed
for
lysosomes
organelles.
normal
250
I .4%
the
enzyme-
Cells
substrate
buffer)
were
in
or
heated
incubation.
acetate
and
knife,
stained
101 electron
control
the
in a
Cells
in veronal
embedded
cells
were
acetate
in
briefly
with
microscope.
Epon.
lead
HL-24577
Contract
accepted
to Dorothy
RESULTS
marrow
Megakaryocytes
nucleated
cells
tures
of MK
Laboratory.
University
of Califor-
Research
W-7405-ENG-48.
October
21, 1981.
F. Bainton,
of California
School
M.D.,
Depart-
of Medicine,
constitute
0.l%-0.5%
of the
of the bone marrow.
The general
feamaturation
in human
bone
marrow
resemble
those described
for
tion in this cell line involves
and AM-10486
and Environmental
Inc.
in 0.2
of
or
of patients
from
Calif
Grants
of Health
ofEnergy
rib fragments
Lawrence
by USPHS
© 1982
472
hyde
disease’4
this conenzymes
7.4,
containing
anticoagulants
were fixed
markers.
In the
evidence
that
two
lysosomal
expressed
of Pathology,
San
by the Office
from
were
Division,
pH
fixative
lysosomes
analyses
was
were
the
platelet
secreted
by pressure
heparin
1.5%
profiles.
reaction.
or without
either
that
that
are
in about
examined
they
in
Golgi
containing
constitute
that
vein
at
found
of platelet
other
believe
fixative
plasma
was
cells
release
10 mm
1 5%
from
antecubital
for
buffer,
conclude
contents
When
fixative.
fixative
We
we
syringe,
platelet-rich
platelets
(ACD)
blood)
centrifuged
and
the
acid-citrate-dextrose
distinct,
also
about
MK
in the
vesicles
in only
no evidence
into
the
were
all
event,
vesicles
METHODS
collected
the Department
the Department
Submitted
AND
was
Biomedical
and
Supported
and
marrow
was
digestive
and
rat, and that these vesicles
are distinct
from the time of their formation.
MATERIALS
Human
there
AS
distinct
enzyme,
small
be demonstrated
in these
are
whose
acid phosphatase
(AcPase)
(with
B-glyceroand trimetaphosphate
as substrates)
and
(AS),
in small vesicles
similar
to those we
reported
in the
from a-granules
vesicles
they
lysosomal
in similar
whereas
enzymes.
lysosomes,
of platelets
from patients
with storage
pool
and the “gray
platelet
syndrome”5
support
clusion
since
these
platelets
have lysosomal
absent
a-granule
we present
cytochemical
and
platelets
contain
containing
both
aggregation
indicate
that
Furthermore,
or
of
MK,
study’2
that
rat megakaryocytes
(MK)
and platelets
store lysosomal
enzymes
in vesicles
distinct
from both
dense
bodies
and a-granules.
Recent
cell fractionation
but
reduced
present
article,
human
MK
found
size,
organelles.’
not
containing
for
Since
it could
profiles,
were
any
localized
Vesicles
a-granules
that
Another
was
platelet
variable
morphologically
studies
of human
platelets
also
and a-granules
are separable.’3
of
negative
and
Golgi
a-granules.
(AS),
In mature
of
and many of their components
have been
Although
a-granules
are often
considered
somes,
we concluded
from
a previous
than
of all stages;
The
Heavy
vesicles;
MK
AcPase
Acid
in those
F. Bainton
smaller
complex.
region
of man and other
a variety
and
confined
lightly
PLATELETS
contain
as
but
25%
marrow
maturation.
coated
limited
to
(MK)
Golgi
Dorothy
arylsulfatase
localiza-
bone
in the
as well
and
order
cytochemistry.
cytoplasmic
usually
cisternae
B
human
present
cisternae
In
subcellular
and
MK,
of
is
megakaryocytes
normal
microscopy
recognizable
considerable
and
in
and
however,
disagreement.
enzymes
(AcPase)
youngest
some
platelets
“lysosome,”
Megakaryocytes
Platelets
E. Bentfeld-Barker
in human
of the
of appearance
by electron
phosphatase
the
of
lysosomal
and
enzymes
identity
in Human
of
ploidy and the development
mic organelles;
electron
cytoplasmic
maturation
other
species.’9
Maturaincreasing
nuclear
poly-
of a unique
microscopy
begins
during
set of cytoplashas shown
that
the ploidization
phase,
but continues
after
nuclear
development
is
complete.
Since the two processes
are neither
synchronous nor sequential,
classification
of the maturational
stages
of MK is difficult.
The nomenclature
used in the
Blood,
Vol. 59.
No. 3 (March),
1982
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MEGAKARYOCYTES
AND
PLATELET
LYSOSOMES
473
Fig. 1 .
Golgi region of an immature
human
MK tested
for AcPase.
Several
stacks of Golgi cisternae
(Gc) are evident.
In most atacks,
reaction
product
is confined
to one cisterns
(arrows)
and nearby coated vesicles
(cv), while the remaining
cisternae
show little or no
reaction.
No typical a-granules
can be identified
in this field; many of the mitochondria
(m) have been disrupted
by cold fixation
or the acid
incubation
medium.
Tissue fixed in 1 .5% glutaraldehyde
for 30 mm at 4’C and incubated
in the Barks-Anderson
medium
for 3 hr at 30”C (n,
nucleus;
nu, nucleolus;
x22,000).
Lower Inset: Higher
magnification
of part of same cell. Reactive
and unreactive
coated vesicles
can be
identified
( x 36,000).
Upper Inset: Light micrograph
of 1 m Epon section
of an immature
MK similar
to the cell shown
in the electron
micrograph.
It has scant cytoplasm
and a bibbed
nucleus
with several
large nucleoli
( x2000).
heavy
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BENTFELD-BARKER
474
present
study
is based
on
the
system
proposed
Paulus2#{176}and used in our previous
paper
on rat
Briefly:
(1) “immature
MK”
are small (-15-18
diameter)
cells
that
have
a nucleus:cytoplasm
greater
than
I and few cytoplasmic
organelles
than
MK”
large
are
nucleus,
rough
numbers
of polyribosomes;
heterogeneous
in size,
a well
endoplasmic
lobes
are
fully granulated
Immature
a
“maturing
lobulated
developed
Golgi
complex,
abundant
reticulum
(RER),
and
variable
numbers
of a-granules
(3) “mature
MK”
are
nuclear
(2)
have
by
MK.’2
sm in
ratio
other
and
also
closely
and
cells
demarcation
heterogeneous
apposed,
divided
constitute
into platelet
a minority
is
fields.
of the
total
MK population,2’
and are very difficult
to identify
morphological
criteria,
especially
in thin sections.
youngest
cell we identified
by electron
microscopy
by
The
(not
often
present
is shown
same
localized
cytoplasm.
can
in Fig.
stage
4A.
in vesicles
At higher
be
seen
that
the
MK;
tion
therefore,
for AS
the
(Fig.
Golgi
can
region,
be
is depicted
identified
(-80-120
as
nm)
inset).
that
Reaction
containing
in Fig.
small
are
I
Forming
.
perinuclear
is also
(Figs.
region
cation
a small
cisternae.
cisterna,
2 and
3) have
with accumulating
membranes,
and
is
lysosomes
vesicles
(Fig.
evident
in each
amount
The
mature
variable
vesicles
and
a-granules
may
RER,
unreactive.
a large
well
a-granules,
well developed
were
never
1, lower
probably
reticulum,
Golgi
equivalyso-
be present
including
Maturing
in
the
MK
developed
Golgi
vesicles,
demarRER.
As can be
seen in Fig. 3A and B, they showed
a similar
tion of AcPase.
In favorable
sections,
reaction
was found in vesicles
and a few Golgi cisternae,
RER
to the
coated
stack,
chiefly
in one or two cisternae,
lent to GERL
(Golgi,
endoplasmic
some22),
although
some of the other
nuclear
limited
AcPase-positive
usually
product
more
activity
reactive.
distribuproduct
but the
In
more
cells,
the AcPase-positive
vesicles
were
more
in size ( 1 75-250
nm), suggesting
that smaller
may fuse to form larger
vesicles.
All appeared
homogeneous
boundaries.
was difficult
showing
no cellular
As we had found
to demonstrate
debris
with rat
in mature
within
MK,
cells
their
AcPase
by the
lead-salt
methods
used for electron
microscopy.
Nevertheless,
a few small reactive
vesicles
could usually
be
seen in thin
sections
of such
cells.
At this stage
of
maturation,
Golgi complexes
considerably
in size and are
trol
preparations
incubated
heated
showed
have generally
decreased
difficult
to sample.
Conwithout
substrate
or
no reaction.
In MK ofall
maturational
stages,
from the youngest
recognizable
cells to the most mature,
AS was always
present,
confined
to small
vesicles.
Although
reactive
illustrates
a cell
Reaction
size
we
of the
reflect
AS in
rats,
Part of
mature
at
product
throughout
(inset,
Fig.
structures
is
the
it
4B),
are
distinct
densities
seen in MK after
4B)
are very
probably
unidentified
cell,
AcPase
4B
AS.
reactive
some
larger
showing
region,
from the a-granules,
which
are unreactive
and usually
larger.
Dense
bodies
(illustrated
in Fig. 5, inset)
are
normally
not seen in routine
electron
microscopy
of
region.
also
for
of variable
magnification
for lysosomal
not be completely
without
substrate,
A slightly
Figure
reacted
product
could
bated
but
Golgi
product
in any component
We presume
that this may
shown)
had a bibbed
nucleus
similar
to that of the cell
shown
in the upper
inset of Fig. 1 . It measured
1 8 tm
in diameter
and showed
AcPase
reaction
in the Golgi
lobes,
in the
BAINTON
limited
sampling,
since we previously
observed
the Golgi
region
of MK from thombocytopenic
which
have a higher
proportion
of young
MK.
the highly
demarcated
cytoplasm
of a typical
MK
the cytoplasm
were
never
saw reaction
secretory
apparatus.
the
membranes;
in size, the
and
vesicles
AND
enzyme.
The
abolished
suggesting
endogeneous
incubareaction
reaction
product
in controls
incuthe presence
of
substrate.
However,
cells heated
in a water
bath for 10 mm at 90#{176}C
prior to
incubation
in complete
medium
were totally
devoid
of
reaction
product,
indicating
enzyme
inactivation;
morphological
preservation
under
these
conditions
excellent.
Vesicles
reactive
for both enzymes
were also
strable
in circulating
platelets.
Figure
5 shows
let from
blood
dripped
directly
into fixative
anticoagulant.
preserved,
body.*
The disc shape
of the
as are numerous
a-granules
Figures
6, 7, and
AcPase,
enzymes
were
AS,
than
8 show
the a-granules.
We
did
bodies
are
characteristic
similar
light
dense
(Fig.
body
platelets
not test
(see
halo
incubated
MK
All
for
three
smaller
for TMPase.
of lysosomes
in platelets
density
of reaction
prodand the platelet
dense
Fig. 5).
is helpful
5, inset).
demona platewithout
platelet
is well
and one dense
and
TMPase,
respectively.
present
in vesicles
somewhat
Cytochemical
identification
poses problems
because
the
ucts
for lysosomal
enzymes
was
When
present,
in identifying
In preparations
the
the
incubated
for AS, we noted
densities
that
we consider
due to
reaction
product
in about
25%
of platelet
profiles.
In
preparations
processed
with our standard
fixation
and
embedding
techniques,
but not incubated
for enzymatic
activity,
we saw the
represent
serotonin-containing
about
7%
incubated
5In
granules;
density,
contents,7
our
of platelet
profiles.
for AS, even if all
earlier
paper’2
although
other
and
densities
dense
this
components
therefore,
Thus,
in
the dense
we identified
these
amine
contribute
appear
we have
may
to form
dropped
that
we
bodies
preparations
bodies
were
organdIes
this
as serotonin
to
a major
assume
in only
their
portion
designation.
electron
of their
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Fig. 2.
Golgi region of a maturing
MK. Surrounding
a centriole
(cc) in the perinuclear
area are numerous
stacks of Golgi cisternae
(Gc)
and abundant
vesicles
similar to, but more extensive
than those seen early in maturation.
The cytoplasm
is replete
with
a-granules
(a),
demarcation
membranes
(dma) and rough endoplasmic
reticulum
(rer) ( x 1 6,000).
Inset ( x 38,000):
Higher
magnification
of a few Golgi
cisternae
with
numerous
adjacent
vesicles
(ye), some of which
are coated
(cv). Occasionally,
a Golgi cisterna
is swollen
and contains
flocculent
material
(arrow).
Fixed
in glutaraldehyde
at 4’C, postflxed
in 0s0,
and stained
en bloc with uranyl
acetate
(n, nucleus).
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BENTFELD-BARKER
476
Fig. 3.
AND
(A)
maturing
Golgi
MK.
which
complex
AcPase
product
is confined
nae (arrows)
and
(cv).
BAINTON
of a
reaction
to two
cistercoated
vesicles
appear
to
bud
from
a
reactive
cisterna.
Alpha
granules
(a)
are
unreactive.
Tissue
prepared as in Fig. 1 ( x 35,000).
Inset:
Light
micrograph
similar
of
maturation
has increased.
toplasm
ratio
compared
a
cell
stage.
at
Cell
a
size
but the nucleus:cyhas decreased
as
with
the
immature
cell
in Fig. 1 (x1500).
(B) Golgi complex of another
maturing
MK. but
at a more advanced
stage of maturation than Fig. 3A. Distribution
of
reaction
product
lar to that
negative
(arrows)
shown
is simi-
in Fig. 3A.
a-granules
(a).
Note
Tissue
prepared
as in Fig. 1 (x33,000).
Inset: Light micrograph
of a similar
cell.
which
exhibits
a Iobult.ted
nucleus
and a greatly
decreased
nucleus:cytoplasm
ratio ( x 1500).
misclassified
would
as
account
When
lysosomes
(which
for less than
platelets
were
30%
incubated
deposits
similar
to those
were found in only about
is unlikely),
of the
total
for
they
cisterna,
lysosomes.
AcPase,
size
dense
seen after
incubation
I 5% of platelet
profiles.
for AS
Since
it was
in both
always
MK
and
present
platelets.
in vesicles
of variable
Unfortunately,
we could
not determine
whether
the same organelles
for both enzymes
or whether
there are two
are reactive
populations
more
limited
experiments
were
performed
with
TMPase,
we have
not attempted
to quantitate
our
results.
With our methods,
it is impossible
to determine
whether
the sites of these
different
enzyme
activities
of reactive
organelles.
Nevertheless,
it is clear
that
a-granules
were not reactive
for either
enzyme.
Previous
cytochemical
studies232t
have reported
AcPase
to be present
in what
appeared
to be platelet
agranules;
however,
this demonstration
usually
required
are the same.
The
smaller
percentage
of AcPasereactive
platelets
may reflect
a narrower
distribution
of this enzyme
as compared
with AS or it may be due
enzyme
distribution.
Furthermore,
micrographs
of platelets25
clearly
to technical
latency.
of the enzyme
outside
of these
investigations
problems,
such
as
partial
cytochemical
platelet
activation
identification
DISCUSSION
These
platelets
studies
provide
and MK contain
that can be designated
shown
to be present
immature
and maturing
in platelets
further
evidence
a distinct
class
primary
in the
MK,
Recent
that human
of organelles
also
lysosomes.
AcPase
was
secretory
apparatus
of
as well as in vesicles
of
mature
MK and some platelets.
As we had found in rat
MK,
heaviest
reaction
product
was confined
to one or
two cisternae
and coated
vesicles
in the Golgi
complex.t
Although
AS was never
found
in any Golgi
tin
GERL.
also
always
reported
our
previous
However,
appeared
possible.
in other
paper’2
since
reactive
other
Golgi
in some
Inasmuch
cell
we designated
types,22
cases,
as similar
and
the
cisternae
reactive
of MK
a clear-cut
uncertainties
we have
done
cisternae
in both
distinction
have
no further
as
or
injury
Fig.
that
studies
platelet
platelet
a-granules.
fractionation
studies
presence
product
have
as well, and none
to MK,
where
is less equivocal
of fractionated
lysosomes
are
than
Although
some
of platelets2728
of lysosomal
enzymes
platelets
distinct
from
of the early
demonstrated
in the
“granule
tion,”
new separation
procedures’3
have made
ble to obtain
a-granule
fractions
of much
purity.
Such
fractions
show
little
lysosomal
but
rather
are
rich
in fl-thromboglobulin,
factor
platelet
altered
some
published
show a distribution
4).
biochemical
indicate
may
the a-granules
was extended
of reaction
(see
that
cell
the
frac-
it possigreater
activity,
platelet
4, platelet
growth
factor,
and fibrinogen.’#{176}29 The
lysosomal
enzymes
that
have
been
studied
species
was
now
not
been
cytochem-
ical tests to define
reactive
sites
GERL
in MK more precisely,
as Golgi-associated
cisternae.
we here
refer
to the
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MEGAKARYOCYTES
AND
PLATELET
LYSOSOMES
477
Fig. 4.
(A) Mature
human MK. Note the numerous
a-granules
(a) and demarcation
system (dms) ( x 1 7,000).
Inset: Light micrograph
of
a mature
MK showing
morphology
typical of a thrombogenic
or mature
MK. The nuclear lobes are close together
at one pole of the cell, and
the voluminous
cytoplasm
appears
to be fragmenting
( x 1000).
(B) Mature
human MK processed
for AS. The cytoplasm
is replete
with
channels
of the demarcation
system
(dms). Numerous
a-granules
(a) are also present.
AS is found in vesicles
of variable
size (ly), but
usually somewhat
smaller
than the a-granules,
which are unreactive.
Tissue fixed in paraformaldehyde-glutaraldehyde
for 30 mm at 4’C
and incubated
for 2 hr at 250’C
in Goldfischer’s
medium
for AS. Inset: Higher
magnification
of small portion
of another
MK showing
numerous
unreactive
a-granules
and two lysosomal
vesicles
(ly) containing
AS reaction
product
( x 17,000).
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
BENTFELD-BARKER
478
AND
BAINTON
Fig. 5.
a-granules
Human
platelet.
The
(a) vary
in size and
density,
and
some
contain
nucleoids
(small arrows).
The core of
the dense body (d) nearly fills the
membrane-bound
vesicle.
A much
more prominent
halo is frequently
seen
(inset).
Fixation
at 37’C
in
1 .5% glutaraldehyde
without
anticoagulant
(m. mitochondrion;
mt,
microtubules)
( x24,000;
inset,
x 52.000).
Fig. 6.
Human
platelet
incubated for AS. In this specimen
the
a-granules
(a)
are
clearly
seen
with
their characteristic
nucleoids
(small arrows).
Two densities
(ly)
probably
represent
AS reaction
product;
the third (d) a dense body.
Tissue
fixed
hyde
Fig.
and
48
in
I .5%
further
glutaralde-
processed
as in
(x24,000).
Fig. 7.
Human
platelet
incubated
for AcPase.
Two
densities
are seen: one (Iy) is considered
to
be due to lead
phosphate
and
hence
represents
a lysosome.
while
the other
(d) is probably
a
dense body
with its characteristic
clear
halo
(a,a-granule).
Tissue
prepared
as in Fig. 1 ( x24,000).
Fig. 8.
Human
platelet
incubated
for
demonstration
of
TMPase.
This enzyme,
which
prosumably
also.”
(ly)
demonstrates
is localized
identical
lysosomes
in small
to
those
vesicles
showing
AcPase and AS activity.
Fixation
as
in Fig. 48; incubation
for 90 mm at
30CC
sediment
in fractions
of lower
shown
by electron
microscopy
of mitochondria
and
membrane
that
these
vesicles
probably
enzymes
found
in that fraction.
try has
tions.
not
In clinical
yet
been
performed
investigations
pool
deficiencies,
described
whose
dense
bodies,
but
density,
which
have been
to be composed
mostly
vesicles.’3
contain
However,
on
the
of patients
We
the
believe
lysosomal
cytochemis-
various
with
frac-
storage
at least
one
patient
has
been
platelets
lack
both
a-granules
and
contain
normal
amounts
of lysosmal
enzymes.’4
described
other
both
The
“gray
platelet
in I 97 1 and since then
patients,’5’3’
platelets
all, while
(x24,000).
is another
syndrome,”
first
identified
in several
disorder
in
which
the
contain
either
very few a-granules
or none
the platelet
lysosomal
enzymes
are normal
biochemical32
and cytochemical
“experiments
of nature”
also point
somal
organdIes
in platelets.
criteria.’5
to distinct
at
by
These
lyso-
Studies
of secretion
kinetics
indicate
that thrombin,
but not ADP,
causes
platelets
to secrete
a certain
amount
of each
lysosomal
enzyme,
generally
about
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
MEGAKARYOCYTES
AND
PLATELET
50%.
acid
3-glycerophosphatase
Although
be distributed
other enzymes
Accordingly,
two
in the
tested,
same
only
Fukami
classes
and
of platelet
glycerophosphatase,
This
could
of $-glycerophosphatase
Unlike
form
a
granules,34
similar
vesicles
to those
of variable
subcellular
about
10%
Salganicoff’
the other
is certainly
also reflect
to the
the primary
population
MK and
of
479
appears
lysosomes:
and
somal
enzymes.
secretion
kinetics
LYSOSOMES
one
the
marker
subsequent
studies
immunocytochemical
have
in our
postulated
containing
remaining
filyso-
but the
binding
membrane.
lysosomes
of leukocytes,
which
of morphologically
identifiable
platelet
lysosomes
appear
to be
most
other
cell types.
They
are
size, which
apparently
are pinched
off from certain
Golgi cisternae,
perhaps
GERL
membranes.35
Compared
with ct-granules,
they constitute
a
minor
proportion
of the MK or platelet
organelles,
and
they
probably
are not recognizable
without
specific
cytochemical
stains.
Their
contents
(without
the cytochemical
stains)
may have an electron
density
similar
to that of a-granule
contents;
however,
especially
as
seen
in MK,
they
are in no way
identical
to a-
On
is known
about
the basis
of
osmium-fixed
in the Golgi
tissue,
complex.
the formation
morphological
Jones36
proposed
However,
since
ofa-granules
observations
clear
that
maturation.
immature
been
found,
(Halverson
results
of
We hope that
being developed
et al.37)
formation
relationship
the
will
soon
make
ofa-granules
and
to lysosomes.
lysosomes
are present
very early
A few a-granules
may also be
MK, but they become
more prominent
it
thus
It is
in MK
seen
in
later
in maturation.
The
lysosomes
platelets
digestive
of both
rat
show none of the debris
activity
that
is normally
lysosomes.
We therefore
believe
lysosomes
in both cells. Although
have attempted
to demonstrate
lets,38#{176} most
appear
to be
canalicular
platelets
tion
of
has
have
taken
system.
never
platelet
be extracellular)
current
diagrammed
Cytochemically,
Dense
body
concluded
up are
human
enzymes
is still
unknown.4’
of the
they
lysosomes
are
small
and
of
that they are primary
several
investigators
phagocytosis
in platesubstances
retained
and
(which
in
that
the
material
by
the funcappears
organelles
by ultrastructural
the diagram
in
labeled
MK
or other
evidence
seen in secondary
that
actually
lysosomal
concept
and
and
Digestion
ofexogenous
been demonstrated,
platelets,
as revealed
is summarized
in
in
in
that they arise
no cytochemical
yet
have been equivocal.
procedures
now
to follow the
clarify
their
Our
Little
has
laboratory
possible
further
granules.
MK.
for them
fractions
as the
of it is secreted.
possible,
differential
lysosome
to
to
of human
cytochemistry,
Fig. 9. We have
as distinct
entities.
acid-hydrolase-con-
I
Surface
connected
Canalicular
System
tubular
IX
membrane
Fig.
9.
Diagram
of
a human
platelet
showing
components
visible
by
electron
microscopy
and
cytochemistry.
In addition
to
membranous
components.
mitochondria,
microtubules.
and glycogen.
we have illustrated
four types
of storage
organdIes:
a-granules,
dense
bodies,
lysosomes,
and peroxisomes.
While
the first two
can
be Identified
morphologically,
their
nomenclature
has been
inconsistent
and confusing.
Of the original
terminology
adopted
for platelet
organeIIes,
only “a-granulomere,”
modified
to “a-granule”
has survived
and is now the almost universally
accepted
designation
for the most prominent
platelet
granule.
Likewise,
dense bodies
have
been called
various
names,
none of which,
including
the recently
proposed
“-granule,”4
is indicative
of their chemical
nature.
In view
of
the widespread
use of the term
“dense
body,”
we suggest
it be retained
until a more definitive
name is devised.
Peroxisom.s
and
Iysosomes
are recognizable
only by cytochemical
stains.
We have included
peroxisomes.
although
thus far only one typical
peroxisome
component.
catalase,
has been demonstrated
in MK and platelets
of normal
individuals.TM
Platelct
lysosomes
are small vesicles
without
a
recognizable
“granule”
morphology.
Although
the term “X-granule”4
emphasizes
their individuality,
we consider
it superfluous;
platelet
lysosomes
are similar
to primary
lysosomes
of other
cells, for which
no other
name has proven
necessary.
From www.bloodjournal.org by guest on June 16, 2017. For personal use only.
BENTFELD-BARKER
480
taming
somes
vesicles
of most
AND BAINTON
ACKNOWLEDGMENT
that
appear
similar
to primary
lysoother
cell types.3435
On the basis of the
of evidence
discussed
above,
we believe
We
several
lines
that they should
be recognized
as separate
organelles
and not be considered
a subpopulation
of a-granules.
wish
technical
for
to thank
Yvonne
assistance,
preparation
of
prepared
by Bob
19.
Breton-Gorius
Jacques
as well
the
as
and
Barbara
Ivy
Hsieh
for
and
Susan
Turner
(Fig.
9)
Poetter
manuscript.
The
diagram
excellent
was
Stevens.
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1982 59: 472-481
Identification of primary lysosomes in human megakaryocytes and
platelets
ME Bentfeld-Barker and DF Bainton
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