The effects of cell age on protein synthesis were examined in the nucleated red blood cells of rainbow trout (Oncorhynchus mykiss). Total DNA content was unaffected by cell age, whereas total RNA content in young red blood cells was roughly ten times as high as that in old red blood cells. The mRNA levels for haemoglobin, carbonic anhydrase and the chloride/bicarbonate (Cl(−)/HCO(3)(−)) exchanger were also approximately tenfold higher in young red blood cells. Although young red blood cells synthesized roughly five times more protein under steady-state conditions, total protein concentration was not affected by cell age. Despite large reductions in mRNA levels with red blood cell ageing, the concentrations and/or activities of the respiratory proteins were largely preserved. In contrast, the ability to mount a heat shock response was greatly reduced in older red blood cells. Young red blood cells produced 13 times more heat shock protein 70 mRNA following heat shock and four times more 70 kDa protein after recovery. They also transcribed much more heat shock cognate 71 and heat shock factor mRNA than did older red blood cells under steady-state conditions.

Becker
J.
,
Craig
E. A.
(
1994
).
Heat-shock proteins as molecular chaperones
.
Eur. J. Biochem
219
,
11
–.
Betke
K.
,
Savelsberg
W.
(
1950
).
Stufenphotometrische hämoglobinstimmung mittels cyanhämoglobin
.
Biochem. Z
320
,
431
–.
Boutilier
R. G.
,
Ferguson
R. A.
(
1989
).
Nucleated red cell function: metabolism and pH regulation
.
Can. J. Zool
67
,
2986
–.
Chomczynski
P.
,
Sacchi
N.
(
1987
).
Single-step method of RNA isolation by acid guanidinium thiocyanate—phenol—chloroform extraction
.
Analyt. Biochem
162
,
156
–.
Cohen
N. S.
,
Ekholm
J. E.
,
Luthra
M. G.
,
Hanahan
D. J.
(
1976
).
Biochemical characterization of density-separated human erythrocytes
.
Biochim. Biophys. Acta
419
,
229
–.
Currie
S.
,
Tufts
B. L.
(
1997
).
Synthesis of stress protein 70 (Hsp70) in rainbow trout (Oncorhynchus mykiss) red blood cells
.
J. Exp. Biol
200
,
607
–.
Currie
S.
,
Tufts
B. L.
,
Moyes
C. D.
(
1999
).
Influence of bioenergetic stress on heat shock protein gene expression in nucleated red blood cells of fish
.
Am. J. Physiol
276
,
990
–.
Denton
J. E.
,
Yousef
M. K.
(
1975
).
Seasonal changes in hematology of rainbow trout, Salmo gairdneri
.
Comp. Biochem. Physiol
51
,
151
–.
Härdig
J.
(
1978
).
Maturation of circulating red blood cells in young Baltic salmon (Salmo salar L.)
.
Acta Physiol. Scand
102
,
290
–.
Hartl
F. U.
(
1996
).
Molecular chaperones in cellular protein folding
.
Nature
381
,
571
–.
Henry
R. P.
,
Tufts
B. L.
,
Boutilier
R. G.
(
1993
).
The distribution of carbonic anhydrase type I and II isozymes in lamprey and trout: possible co-evolution with erythrocyte chloride/bicarbonate exchange
.
J. Comp. Physiol
163
,
380
–.
Hevesy
G.
,
Lockner
D.
,
Sletten
K.
(
1964
).
Iron metabolism and erythrocyte formation in fish
.
Acta Physiol. Scand
60
,
256
–.
Houston
A. H.
,
Murad
A.
(
1991
).
Hematological characterization of goldfish, Carassius auratus L., by image analysis: effects of thermal acclimation and heat shock
.
Can. J. Zool
69
,
2041
–.
Hubner
S.
,
Michel
F.
,
Rudloff
V.
,
Appelhans
H.
(
1992
).
Amino acid sequence of band-3 protein from rainbow trout erythrocytes derived from cDNA
.
Biochem. J
285
,
17
–.
Keen
A. M.
,
Steele
C.
,
Houston
A. H.
(
1989
).
The circulating erythrocytes of rainbow trout
.
Comp. Biochem. Physiol
94
,
699
–.
Laemmli
E. K.
(
1970
).
Cleavage of structural proteins during the assembly of the head of bacteriophage T4
.
Nature
227
,
680
–.
Lane
H.
(
1984
).
Nucleoside triphosphate changes during the peripheral lifespan of erythrocytes of rainbow trout (Salmo gairdneri)
.
J. Exp. Zool
231
,
57
–.
Leary
S. C.
,
Battersby
B. J.
,
Hansford
R. G.
,
Moyes
C. D.
(
1998
).
Interactions between bioenergetics and mitochondrial biogenesis
.
Biochim. Biophys. Acta
1365
,
522
–.
Lee
Y.-K.
,
Manalo
D.
,
Liu
A. Y.-C.
(
1996
).
Heat shock response, heat shock transcription factor and cell ageing
.
Biol. Signals
5
,
180
–.
Liu
A. Y.-C.
,
Lin
Z.
,
Choi
H.-S.
,
Sorhage
F.
,
Li
B.
(
1989
).
Attenuated induction of heat shock gene expression in ageing diploid fibroblasts
.
J. Biol. Chem
264
,
12037
–.
Mestril
R.
,
Dillman
W. H.
(
1995
).
Heat shock proteins and protection against myocardial ischemia
.
J. Mol. Cell. Cardiol
27
,
45
–.
Morimoto
R. I.
(
1993
).
Cells in stress: transcriptional activation of heat shock genes
.
Science
259
,
1409
–.
Murphy
J.
(
1973
).
Influence of temperature and method of centrifugation on the separation of erythrocytes
.
J. Lab. Clin. Med
82
,
334
–.
Nadeau
K.
,
Das
A.
,
Walsh
C. T.
(
1993
).
Hsp 90 chaperonins possess ATPase activity and bind heat shock transcription factors and peptidyl prolyl isomerases
.
J. Biol. Chem
268
,
1479
–.
Pahlavani
M. A.
,
Harris
M. D.
,
Moore
S. A.
,
Weindruch
R.
,
Richardson
A.
(
1995
).
The expression of heat shock protein 70 decreases with age in lymphocytes from rats and rhesus monkeys
.
Exp. Cell Res
218
,
310
–.
Parsell
D. A.
,
Lindquist
S.
(
1993
).
The function of heat-shock proteins in stress tolerance: degradation and reactivation of damaged proteins
.
Annu. Rev. Genet
27
,
437
–.
Perry
S. F.
(
1986
).
Carbon dioxide excretion in fish
.
Can. J. Zool
64
,
565
–.
Perry
S. F.
,
Gilmour
K.
(
1993
).
An evaluation of factors limiting carbon dioxide excretion by trout red blood cells in vitro
.
J. Exp. Biol
180
,
39
–.
Peterson
R. E.
,
Tu
C.
,
Linser
P. J.
(
1997
).
Isolation and characterization of a carbonic anhydrase homologue from the zebrafish (Danio rerio)
.
J. Mol. Evol
44
,
432
–.
Phillips
M. C. L.
,
Moyes
C. D.
,
Tufts
B. L.
(
2000
).
The effects of cell ageing on metabolism in rainbow trout (Oncorhynchus mykiss) red blood cells
.
J. Exp. Biol
203
,
1039
–.
Rabindran
S. H.
,
Giorgi
G.
,
Clos
J.
,
Wu
C.
(
1991
).
Molecular cloning and expression of a human heat shock factor, HSF 1
.
Proc. Natl. Acad. Sci. USA
88
,
6906
–.
Sekhon
S. S.
,
Beams
H. W.
(
1969
).
Fine structure of the developing trout erythrocytes and thrombocytes with special reference to the marginal band and the cytoplasmic organelles
.
Am. J. Anat
125
,
353
–.
Speckner
W.
,
Schindler
J. F.
,
Albers
C.
(
1989
).
Age-dependent changes in volume and haemoglobin content of erythrocytes in the carp (Cyprinus carpio L.)
.
J. Exp. Biol
141
,
133
–.
Stabeneau
E. K.
,
Vanoye
C. G.
,
Heming
T. A.
(
1991
).
Characteristics of the anion transport system in sea turtle erythrocytes
.
Am. J. Physiol
261
,
1218
–.
Tooze
J.
,
Davies
H. G.
(
1963
).
The occurrence and possible significance of haemoglobin in the chromosomal regions of mature erythrocyte nuclei of the newt Triturus cristanus cristanus
.
J. Exp. Biol
141
,
133
–.
van Kampen
E. J.
,
Zijlstra
W. G.
(
1961
).
Standardization of hemoglobinometry. II. The hemoglobincyanide method
.
Clin. Chim. Acta
6
,
538
–.
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