From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
The
after
Maturation
In
Vitro
By P. A.
Labeling
YNTHESIS
OF
continues
in
with
were
entire
about
(a)
to
lifespan
the
various
E.
ScHIFFER
be
can
tritiated
about
of
Reticulocytes
G.
CRONKiTE,
Hp.
85
pcr
Amino
Acids
GIAcow
ScHNAPPAUF
cent
of which
is hemoglobin,1’2
blood
reticulocytes
as long
as polyribosomes
are
demonstrated
by autoradiography
after
incubating
amino
acids
(AA).5
The
objectives
of this
study
show
that
tritiated
of the red cell
lifespan
P.
of
Tritiated
AND
PROTEIN,
peripheral
Synthesis
present.3.4
reticulocytes
M.
Fate
with
STRYcKMANS,
L.
S
and
and
amino
(b)
reticulocytes
acids
remained
to investigate
incorporated
some
current
their
to
and
relation
mature
for the
concepts
red
cells
of
sizes.
METHODS
Eleven
male Sprague-Dawley
rats and one dog were studied.
Four rats, 220-245
grams,
put in a low pressure
chamber
at a simulated
altitude
of 18,000
feet. Four rats,
410-565
grams, received
phenylhydrazine
(PHH).
Three
rats, 240-265
grams, were untreated
controls.
For details see Table 1. Two additional
rats were
used
for additional
controls as described
below.
There was no bartonellosis
in the rat colony.
When
maximum
reticulocytosis
was expected,
the animals were anesthetized
with intraperitoneal
thiamyal
sodium
(Surital#{174}), supplemented,
when
necessary,
with
ether
by inhalation.
Four to 8 ml. of blood (10-37
per cent of the calculated
blood volume)
was
drawn
into a heparimzed
syringe
from the jugular
vein. The blood was mixed with 4 tCi/ml.
of 3H-histidine#{176} (1 Ci/mMole)
and 4 Ci/mL
of 3H-leuclne#{176} (1.25 Ci/mMole),
and incubated for 90 minutes
at 37 C. A solution
of nonradioactive
histidine
and leucine
(about
100 times
the concentration
of tritiated
leucine
and histidine)
was
added
to the sample
which
was then centrifuged
at 1,500 rpm for 15 minutes.
The supernatant
was discarded
and the cells resuspended
in another
aliquot
of the nonradioactive
AA. After centrifugation
and resuspension
in the nonradioactive
AA, the cells were injected
into the jugular
vein of
the donor animal. Serial blood samples
were
taken
from
the tail for Giemsa
stained smears,
reticulocyte
stain,
and red cell counts for 70 days. Reticulocyte
preparations
were stained
were
#{176}Schwartz
Bioresearch,
From
Medical
the
Orangeburg,
Research
New
Center,
York.
Brookhaven
National
Laboratory,
Upton,
New
York
11973.
Research
First
supported
by the U. S. Atomic
submitted
January
10, 1967;
A. STRYCKMANS,
M.D.:
Institut
Energy
Commission.
accepted
for
publication
July
5, 1967.
Pixmix
Jules Bordet,
1 Rue Heger-Bordet,
Brussels,
Belgium, Research
Scholar, Eastern Leukemia
Society. EUGENE
P. CRONKITE,
M.D.:
Chairman,
Medical
Department,
Medical
Research
Center,
Brookhaven
National
Laboratory,
Upton,
N. Y. 11973.
GIUSEPPINA
GIACOMELLI,
PH.D.:
Labdratorio
RadloLsotopi
di Fisica,
Via
Irflerio
46, Bologna,
Italy. LEwIs
M. Scmmi,
M.D.:
Acting
Head,
Division
of Hematology,
Medical
Research
Center,
Brookhaven
National
Laboratory,
Upton,
N. Y. 11973.
HANSPETER
SCHNAPPAUF,
D.V.M.:
Veterlnar-PhyslologLsches
Institut,
Justus
Lleblg
Universit#{228}t,63 Ciessen,
Frankfurterstra&se
94, Germany.
BLOOD,
VoL.
31,
No.
1
(JANUARY),
1968
33
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
34
STRYKMANS
ET
AL.
C/)
-j
-J
w
0
0
0
2
w
P1
z
0
G)
Ui
-J
Ui
>
z
4
-J
30
C)
20
z
I-
z
Ui
0
C
-I
0
I0
Ix
LU
a-
0(1)
-J
L&JJ
>c00
0
Ui
m
080
40> z
x
C)
w Ifl
w<60
30?
z
C)
01-40
20c
I-j
0
z
-I
LLJO
20
I0
Ui0
a-
DAYS
AFTER
INJECTION
OF
LABELED
RETICULOCYTES
Fig.
(0)
1.-Top:
of the red
The per cent
of labeled
cells in one dog versus
same
sample.
Bottom:
The per
and the mean
grain
the same
cent
count
of the
(0)
red
time.
cells
S.D.:
maximal
value
of
in rat 5 versus
time.
(.)
From
and the mean
grain
5 separate
counts
circulating
labeled
S.D.:
5 separate
From
red cells
counts
and
new
(.)
on
sample.
to precipitate
the reticulum,
destained
with methyl
using
NTB2 Kodak
liquid
emulsion
diluted
in an equal
of twice-distilled
water.
Slides were
exposed
for 20 days at 4 C, developed,
then
with Giemsa
in sodium
phosphate-citric
acid buffer at pH6.
with
count
on the
methylene
autoradiographs
blue
made
alcohol,
volume
stained
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
MATURATION
AND
OF
FATE
35
RETICULOCYTES
00
Ls
0
80
60
1W
I--I
W
Ls.
0<
Zj
UJ<
40
#{149}
RAT
7:5%
20
RAT
OF-
co
\
RETICULOCYTES
o RAT 8:8%
6 RAT 10:100/0
RETICULOCYTES
RETICULOCYTES
12:15%
RETICULOCYTES
Li,-
0
0
0
DAYS
Fig.
versus
2.-The
the time
ticulocyte
levels
20
30
AFTER
INJECTION
40
per cent of the maximal
after the autotransfusion
ranging
from
the control
animals,
too few
survival
curves, a limitation
of
lated animals
was counted
using a
were counted
in the small square.
In
Age in
Months
Stimulus
Phenyl-
Rat
No.
2
14
Rat
No.
3
14
Rat
No.
5
14
Rat
No. 16
Rat
No.
7
Hydrazine
4
2
Miller
eyepiece.
Cells with more
2
Pressure
Rat
No. 10
2
Chamber
8
12
12
0.9 mg X 2,8 and
3 days before
exper.
7
12
12
3.5 mg X 1,7 days
before
experiment
4
30
52
3.5 mg X 2,8 and
3 days before
exper.
7
27
32
19
4
Rat
No. 21
4
Rat
No.22
4
Dog
?
4 days
hrs. before
to
‘
6
8
8
5
10
10
4
15
15
5.5
<1
4.5
<1
5.5
<1
250
1.8
12
exper.
From
5 days
to 2
hrs. before
exper.
2
None
Phlebotomy
Peak
Reticulo..
cyte %
1.8 mg X 2,8 and
3 days before
exper.
From
No.
No. of cc Reticulocytes
Blood
at Day of
Labeled
Experiment
From 6 days to 18
hrs. before
exper.
8
Rat
1
Dose/100
gin
Body
Weight
and Time
From 6 days to 18
hrs. before
exper.
No.
RatNo.12
were
seen to obtain
reliable
red
cent of labeled
cells in the stimuOn each slide, at least 1,000 red cells
than 5 grains were counted
as labeled
Low
Rat
70
CELLS
labeled
reticulocytes
the method.
The per
Table
Animal
60
LABELED
value
of total circulating
labeled
red cells
of labeled
reticulocytes
in 4 rats with recent
after exposure
to low oxygen
pressure.
5 to 15 per
cell
50
OF
240
before
cc, 2 days
experiment
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
36
STRYKMANS
ET
AL.
00
Ls.
U
Ij
0-J
Li,-
jF-
tO
a.
20
30
DAYS AFTER
40
50
INJECTION
OF
60
LABELED
70
RETICULOCYTES
Fig.
3.-The
per cent of the maximal
value
of the total circulating
labeled
red
cells
versus
time after autotransfusion
of labeled
reticulocytes
in 4 rats with reticulocyte
levels
ranging
from
12 to 52 per cent after
injecting
various
doses of phenylhydrazin.
(see below).
The absolute
number
of circulating
labeled
red cells was calculated
for each
sample
using a blood volume
estimated
from the body weight
at the time the sample
was
taken.6
One rat with 10 per cent reticulocytes
was injected
intravenously
with 10 LLCi 8H-histidine
and 10 pCi 3H-leucine
diluted
1/100
with the same nonradioactive
AA. Since no red cells
had more than 3 grains
during
the 4 days of this study,
significant
in vivo labeling
by unincorporated
tritiated
AA is unlikely.
Also, the probability
of significant
in vivo reutilization
of labeled
material
seems
remote
because
no rat had
labeled
reticulocytes
with
more
5 grains
after 50 hours
postinjection.
Furthermore,
found
in a rat which
had had 4 ml. of its blood
than
no cells with more than 5 grains
were
(with 50 per cent reticulocytes
after PHH)
labeled
and then autotransfused
after destruction
of the cells by freezing
and thawing.
Since none of the above
control
slides had more than 5 grains
over a cell, we counted
any
cell with 6 or more grains as being labeled.
For one control
animal (No. 22) and two highly
stimulated
animals
(No. 5 and No. 16),
cell images
were projected
on a screen
to an enlargement
of about 2,000 times. The projected
areas of the first 100 labeled
red cells encountered
and of the nonlabeled
red cells
closest to each were measured
with a planimeter
or with a Zeiss particle
size analyzer
TG23.
Distributions
of sizes of the labeled
and unlabeled
red cells were determined
by plotting
the cumulative
percentages
of sizes for each group
on logarithmic
X probability
paper.
Red cells of rat No. 22 were measured
at days 3 and 53 after labeling,
rat No. 5 at days 7
and 53, and rat No. 16 at days 4, 16 and 45.
Table
Rat
Ident.
Number
Reticulocyte
Peak
(per cent)
7,8
5to8
3, 5, 10, 12, 16
#{176}Lifespan of the
ti S.D.
Red
10 to 52
Cells.
2
Mean
Life Span#{176}
(days)
51.2
36.9
j
1
7.2
Mean
Potential
Life Span
(days)
53 ±
48.6
±
i
4.5
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
MATURATION
AND
FATE
OF
j
,I*J_
8
I
I
RAT 22:
-
37
RETICULOCYTES
I
I
I
CONTROL
-
6-
jz;;
:
z
4
-
0
Li
#{149}
WHOLE
>..
;
2
-
3 DAYS
0
co
<
MEDIAN
I
MEDIAN
5
AREA
AREA
lO
3D.
WHlLE
20
CUMULATIVE
POPULATION
OLD
OLD
RED
RED
REDCELLS
30
40
50
PERCENT
RED
CELLS
CELLS
CELLS
09
POPULATION
60
OF
OF
70
-
80
90
MEASURED
00
RED
CELLS
Fig.
4.-The
cumulative
per cent
of the 3-day-old
red cells (0)
to 65-day-old
red cells
(#{149})
in a normal
rat categorized
according
measured
on photomicrographs,
expressed
in arbitrary
units,
plotted
X logarithmic
paper. The bars represent
the range of each area category.
and of the 1to their
area
on probability
RESULTS
Fig.
1 shows
throughout
the
their
constancy
Fig. 2 shows
that
tion-Table
1) did
rats
not
with
10 and 15 per
by day 28. Animals
cent
with
-Table
The
with
lose
5 and
labeled
mean
grain
tT
i’4o
number
of cells
N
=
number
of labeled
was
lifespan,
in the number
represents
estimated
not
as
signfficanfly
lated
animals
(Table
Both
labeled
and
had
levels
lost 17 and 30 per cent respectively
of 12 to 52 per cent
(PHH
stimulus
normal
distributions
tions
overlap
considerably,
larger
at time
surviving
by planimetry.
mean
lifespan
the 50 per cent
different
between
red
but
that
the
of
tritiated
with
of labeled
cells,
the phenomenon
of their
than
(hypoxia
stiinuladays,
whereas
those
cells by 30 days
by:
labeled
cells
2).
unlabeled
have
times
cells
cent reticulocytes
for at least
27
initially
integration
was
performed
show
a significantly
reduced
tential
lation;
red
(Fig.
3).
N#{149}dt
=
decrease
presumably
the
o
N0
minal
paper,
over
count
8 per
cells
reticulocytes
reticulocyte
J
1
T=
is 1.09
the
1) lost 21-55
per cent of the labeled
red
mean
red cell lifespan
(T) was determined
-
The
animals
of
lifespan.
cells
sizes
median
the
(no
strongly
2). The
on this
slightly
longer
area
labeling
a straight
line
of senescence.
4).
nonlabeled
t after
The
(Table
point
the
(Fig.
AA
The
of the
red
stimulated
steep
line
and
or its
heavily
reticulocytes)
sizes
two
labeled
If
extrapostimuon
of the
young,
cell.
ter-
on probability
The
mean
po-
the
day
3
populared
thickness
cell
of
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
38
10
-
8
-
A
-
#{149}
WHOLE
5
1-
I
I
16 AFTER
I
RAT
I
I
OLD
16
DAY
OLD
RED
CELLS
#{176}
45
DAY
OLD
RED
CELLS
RED
CELL
THE
-
45
I
1
1
RED
DAY
I
STIMULATION
AL.
-
CELLS
POPULATION
OLD
RED
AROUND
CELLS
-
10
0
W
I
I
I
I
I
I
I
1
1
1
1
1
I
I
x RAT
RAT
22:
22:
1
OLD
#{149}
RAT
5
53
DAYS
D
RAT
16:
53 DAY
UNLABELED
THE
53 DAY
UNLABELEDCELLS
ERYTHROPOIETIC
UNLABELED
I
I
I
1
LABELED
RED CELLS
RED
CELLS
AROUND
OLD
RED
CELLS
CELLS
1
I
AFTER
STIMULATION
45
DAYS
STIMULATION
ERYTHROPOIETIC
o
I
ERYTHROPOIETIC
DAY
A
w
o
Fr
STRYKMANS
AFTER
5-
Li
<3.
-
-
-
-
*-
-
I
I
5
10
CUMULATIVE
1
20
I
3040
PERCENT
I
1
1
1
506070
OF
MEASURED
1
80
I
90
RED
95
CELLS
Fig. 5.-Top:
The distribution
by size of a labeled
red cell cohort at 4, 16 and
45 days age compared
with the whole normal red cell population
in rat 16.
Bottom:
The cumulative
per cent of different
populations
of red cells categorized
according
to their area.
the young
red cell is increased
by the same factor,
then the ratio of the median
volume
of the labeled
cell on day 3 to the median
volume
of the unlabeled
red
cells is 1.14 (1.16 for the mean volumes).
Fig. 5 (bottom)
suggests
that the labeled
red cells on day 53 are slightly
smaller
than
the whole
red cell population
(control
rat No. 22) and that the
distributions
of the areas of the unlabeled
red cells of rats No. 5 and 16 have
returned
to normal
values by days 53 and 45, respectively.
Fig. 5 (top)
shows
that in rat No. 16 (PHH
stimulated),
the labeled
red
cells on day 4 are considerably
larger
than the whole
population
on day 45,
less so on day 16, and even less so on day 45.
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
MATURATION
AND
FATE
OF
39
RETICULOCYTES
U)
I
I
-J
IIIIlIIIIII’’’’#{149}#{149}
RAT
I
16
4Li
LsJ
>MEDIAN
<MAX.
I-
z
<MEDiIIIIIIIIii
E-
WI0
2
6
10
14
18
22
26
DAYS AFTER
Fig.
6.-The
(0)
size
of cohort
of labeled
In Fig.
34
38
42
46
LABELING
red
expressed
cells
as the
cent
per
of its
versus
time.
This
population
is subdivided
mum,
() > median <maximum,
maximum
30
into
6, the whole labeled
the labeled
red
3 groups:
(a)
53 (>
Max.),
(b)
the
3 categories
(0)
according
See text.
<median.
red cell population
cells larger
than
labeled
red
cells
to
in rat No.
(#{149})
> maxi-
size
16
is
any unlabeled
red
than
the median
smaller
divided
into
cell at day
size
of
the
Median),
and (c) the labeled
red cells
smaller
than
(a) but larger
than
(b) (>Median
< Max).
It can be seen
that group
(a) decreases
faster
than the whole
population,
probably
because
of shrinkage
into groups
(b) and (c) which
increase
at the same time.
Fig. 7 shows
the decrease
with
time
of the per cent of labeled
cells still
containing
reticulum.
Reticulocyte
maturation
time varies
directly
with reticulocyte level.
unlabeled
red
cells
at
day
53
(<
U)
-J
-J
Li
#{176}-#{176}
Li-i
#{176}-#{176}
3 RATS
<
2RATS
5-
2 RATS
1% RETICULOCYTES
8#{176}/o
RETICULOCYTES
32-52%
RETICULOCYTES
00
WIjW
II
-J
0<
40
S.D.
‘S
‘S
‘S
I-
‘S
S.
20
‘S
‘S
0
Li
atO
HOURS
Fig.
in hours
7.-The
after
the
per cent
injection
20
AFTER
30
40
INJECTION
of labeled
of labeled
red
OF
cells
reticulocytes.
LABELED
containing
RETICULOCYTES
reticulum
versus
the time
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
40
STRYCKMANS
ET
AL.
cell
ki-
DisCUssIoN
For
this
technique
the
tracer
netics,
cell,
must
a condition
and
its
due
curves
volume.
We
do
affected
only
No
not
feel
that
cells
by
the
the
first
corrections
the
labeled
Cruz
present
at the
by acetyl
PHH.
and Harris10
have
lifespan
of
loss,
cells which
No direct
all
)
etc.
the factors
The
mean
present
work
that
15
glycine11’12
acute
Iron-59,13
erythropoietic
shown
that
by PHH,
produced
marked
stimulation
this
have
observation
hemorrhage
in response
per
cent
in
decrease
have
is
reticulocytes.
than
lifespan
The
15N-glycine
or
have
evidence
red
but
mean
one
observed
preferential
loss
loss
of
of
red
poietic
cytes
stimulation
probably
have
cytes
ours,
can shrink
Killmann
in order
calculated
cent
mately
larger
20
than
per
the
cent
cells
some
red
is due
to
It
cells.
has
dogs
mean
accelerated
been
shown
normal
into
Nitrogen-
produced
after
Stohlman
has
erythropoiesis
The
animals
present
with
demonstrates
loss
that
of cells,
not
by marked
arises,
then,
random
that
study
than
more
the
loss
of
erythrowhether
loss
greater
or
to
a
erythro-
produces
a more
marked
macrocytosis
in which
the
a shorter
survival.’5
One must
know
how
much
macromacro-
to study
their
survival.
Using
the same
assumption
as
that the volume
of the normal
reticulocyte
is 20 per
average
of
the
cohort
of red cells
to have
a more
later.
method
an
with
cells
stimulates
to
belifespan
the cell
( senescence,
mean
lifespan
of
the first
appeared
is due
that
to distnguish
lifespan.
in
blood
reticulocytes,
The
red
hemoglobin
from surviving
cells.
The mean
potential
lifespan
of the red cells
produced
poietic
stimulation
was normal
in this study.
The question
the
not
precursors
that
produced
time,
ob-
adversely
in
reticulocytes
cell
is decreased
with
been
mature
and that
hemolysis
those
in
into
autoradiographic
59Fe
could
good
labeled
whether
or
volume
the
all
a shortened
or erythropoietin14
to blood
loss
blood
alterations
lifespan.
shown
valid
disap-
using
except
senescence.
all reticulocytes
mature
animals.
and
others
shortening
of lifespan
that the mean
red cell
shows
10
or
count
cells
for
the survival
of
lifespan
is the
suggests
of the
grain
cells
it is important
potential
processes
suggests
normal
red cells in mildly
stimulated
By labeling
a cohort
of reticulocytes
mean
labeled
of injection,
out,
mean
the
uncorrected
moment
affecting
potential
all destructive
exists
that
to
presented
pointed
the
.
escaped
evidence
The
and
span
known
blood
volume
or for possible
changes
due to
technique.
The fluctuations
of
or PHH8
labeling
al.9
red
life
calculated
used
reticulocytes
et
and
last
red
were
were
due
of
the
labeled
weight
be
complete
until
of circulating
the
reticulocyte
the
constancy
day
in body
3 may
because
mean
cells.
for
tension7
during
2 and
PHH
is a result
red
Tribukait.6
Figs.
destroyed
As Beicher
random
increase
red
the
to study
number
with
in
mature
tool
incorporated
from
to low oxygen
and reinjection
by
tween
cells
the total
by
changes
withdrawal
the
red
variation
servations
remain
demonstrated
over the labeled
pear (Fig. 1).
In calculating
are
to be a valid
the
red
initial
cell,
suggesting
volume
occurs
that
a shrinkage
normally.16
of approxiIn
the
present
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
AND
MATURATION
study,
the
on day
labeling
3 still
lation.
does
not
normal,15
but
and maintain
obtained,
The
study
Stohlman
for
for
appeared.
By
cent
decrease
in
out
No.
the
cells,
4 days,
the
median
16,
whole
then
day
4 to 45.
5, top)
shows
red
cell
the
was
cell
age
cell
were
due
of 28 per
cent
the
other
per
cent
from
red
shrinkage
Our
red
day
cell
4 to day
with
age;
judging
findings
are
observed
by
of very
large
produced
soon
circulation.
duction
in
size
on the survival
Since
the
the
after
the
Our
results
during
the
day
only
to a decrease
45,
probably
closer
per
cent.
changes
with
per
200
stimulation
normal
macrocytes
lifespan
of labeled
macrocytes
in normal
methods
used,
autoradiography
in
the
perhaps
maximum
because
maturation
maturation
the
when
time
reticulocytes
erythropoiesis
is
decrease
until
6 or more
hours
high
autoradiographic
threshold
ture
reticulocytes
protein.
The
with
linear
all reticulocytes
only
decrease
enter
the
of the
leave
the
after
the autotransfusion,
used
eliminated
from
a few
after
blood
polyribosomes
6 hours,
at the
same
and have a uniform
maturation
exact
age of the labeled
cells
stant
grain
fragmentation
count
is a mechanism
seen
in
Fig.
of death
still
1.
that
volume)
from
a slow
with
are
animal.
rats was apof reticulowas
greater,
at an earlier
stage
reticulocytes
possibly
consideration
did
because
very
incorporating
AA
as Seip
pointed
out,18
suggests
stage,
remain
in the
blood
not
by senescence.
support
re-
a study
mensuration,
time in the blood.
is known,
one can
1 does
Fig.
removed
reticulocytes
marrow
Labeled
stimulated.
maturation,
Since
the
mean
blood
bone
to
in
conclusion
show
cell
due
normal
agreement
rats.’7
and
red
be
of macrocytic
is rapidly
some
that
of the
only
shown
the
from
shrinkage
their
cent
of the
after
release
has an aver-
in MCHC
and
than
size
population.
The
with
time,
21
can
counts
132
If this
to reality,
as the
grain
disto
occurred
red cell
decreased
same
have
already
down
very time-consuming,
the red cell volume
study
was limited
to one
The maximum
maturation
time
of the reticulocytes
in normal
proximately
33 hours
(Fig.
7). In animals
with
increased
numbers
cytes,
4 still
of the
4 to 45 days
mean
that
almost
114
volume
normal
has
Stohlman’5
indicate
we
1.59 larger
red cells
in
was
original
the
erythropoietic
an
coincidentally
(Fig.
1)
had
cells
by
slight
(more
normal
remains
on day
probably
and
from
of
and
cells
cells
red
of the
decrease
Brecher
to
removed
in the early
days
population
at 4 days still
constancy
red
cells
than
the
population
with
red
labeled
approximately
this
compatible
a cohort
the
cells
by
45,
that
extreme,
per cent larger
of this labeled
popu-
MCHC
occur
count
cells
cell
macrocytes
because
red
of
population
volume
considers
volume
145
cell vioume
normal
of
red
red
red cell and a volume
on a cohort
of labeled
labeled
volume
that
average
of hemoglobin.
average
begun
the biggest
red cells are preferentially
from the bone
marrow,
then the labeled
median
shrinkage
loss
largest
a shrinkage
If one
shows
of the
disappearance
possible
16 (Fig.
day
median
red
that
early
animal
cent
and
loss of hemoglobin
might
MCHC.
The
stable
mean
grain
rules
of the
control
114 per
felt
their
area
1.36 larger
than
average.
Since
sizing
circulation
per
and
on rat
of one
volume
account
shrinkage
a normal
41
RETICULOCYTES
a median
however,
a median
than
the
OF
of reticulocytes
have
Brecher
sizes
FATE
the
say
that
hypothesis
of
not
the
mainto
that
during
the
conthat
From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
42
STRYcKMANS
ET
AL.
Suiy
1. Tritiated
histidine
and leucine
are
incorporated
into hemoglobin
by rat
dog reticulocytes
and fulfill the conditions
of a good
red cell label for
autoradiographic
studies.
2. After mild erythropoietic
stimulation,
all reticulocytes
appear
to mature
into normal
red cells with expected
lifespans.
3. After marked
erythropoietic
stimulation,
the reticulocytes
mature
into red
cells with normal
potential
lifespans
but decreased
mean lifespans.
4. In one animal
studied,
macrocytes
produced
by increased
erythropoietic
stimulation
were found to have the same magnitude
of shrinkage
as do normal
red cells.
5. The normal
maturation
time of the rat reticulocyte
in the blood is about
and
33 hours
in vivo and is increased
following
SUMMARIO
1. Histidina
e leucina
a tritium
erythropoietic
stimulation.
IN INTERLINGUA
es incorporate
ad
in le hemoglobina
de reticulocytos
ratios e canes e satisface
le conditiones
pro un bon marca cellular
in
objectivos
de studios autoradiographic.
2. Post leve grados de stimulation
erythropoietic,
omne le reticulocytos
in normal erythrocytos
con le expectate
tempores
de superviventia.
3. Post
normal
marcate
grados
periodos
viventia.
4. In un
erythropoietic
potential
del
animales
manifestava
de
de
stimulation,
le reticulocytos
superviventia
sed
con
matura
reducite
studiate,
macrocytos
producite
le mesme
magnitude
de
ad
pare
in
maturar
ad
erythrocytos
periodos
de
le
pro
erythrocytos
medie
con
de
super-
per un augmento
del stimulation
restringimento
como
eyrthrocytos
normal.
normal
de maturation
del
33 horas. hUb es augmentate
5. Le tempore
proximativemente
ratios
intra
eyrthropoietic.
reticulocytos
de
post stimulation
le sanguine
es ap-
ACKNOWLEDGMENTS
We wish
Mr. Emil
to acknowledge
the skillful
technical
assistance
of Mrs.
Katherine
Conkling
and
Adaniik.
REFERENCES
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Microsomal
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Giowacki,
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From www.bloodjournal.org by guest on January 21, 2015. For personal use only.
1968 31: 33-43
The Maturation and Fate of Reticulocytes after In Vitro Labeling with
Tritiated Amino Acids
P. A. STRYCKMANS, E. P. CRONKITE, G. GIACOMELLI, L. M. SCHIFFER and HP. SCHNAPPAUF
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