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The O2 dissociation curve of blood of the rhesus monkey (Macaca Mulatta)
Res~irut~on Physiology (1967) 2, 168-172; worth-bollard
Publishing Company, Amsterdam
THE O2 DISSOCIA~ON CURVE OF BLOOD OF THE RHESUS MONKEY (MACACA MULATTA)
J. T.
PARERI
Oregon Regionaf Primate Research Center, Beaverton, Oregon, and Heart Research Laboratory, University of Oregon Medical School, Portland, Oregon, USA
Abstract. The 0~ dissociationcurve of blood of rhesus monkeys was constructed
using blood samples from six animals. The mean 02 tension necessary for SOT! saturation of hemoglobin (Tso) was 32.3 mm Hg at 38 “C and plasma pH of 7.40. This is considerably to the right of the previously published curve for this species. Monkey blood Oxygen affinity of blood Oxygen dissociation curve
Materials and methods Oxygen dissociation curves were constructed with venous blood drawn from six rhesus monkeys into syringes containing fluoride in heparin. No hemolysis was evident. The monkeys were anesthetized with 0.3 to 3.0 mg/kg of phencyclidine hydrocholoride IM (Sernylan, Parke Davis and Co., Ann Arbor, Michigan). Aliquots of blood were tonometered at a temperature of 38 “C with gases of known 02, CO, and N, composition (analysed by the method of Scholander). OZ concentration of the blood was then measured by the method of Van Slyke and Neill, and pH was measured with a Radiometer glass electrode at 38 “C. The technique has been described more fully elsewhere (PARER, HOVERSLAND and METCALFE, 1967). Results and discussion Fig. 1 shows the average 0, dissociation curve, together with the measured points converted by means of a Bohr effect factor of -0.5 units to represent their values at pH 7.40. The measured and calculated data from which the curves were constructed are given in table 1. The CO, tensions and contents are also shown. The average O2 tension at 50% saturation (T,,) ranged from 31-33 mm Hg. The mean 0, capacity was 16.6 vol %; none of the subjects was in an abnormal acid-base status or anemic, Accepted for pablicat~on 21 November 1966. 1 Present Address: Department of Obstetrics and Gynecology, of Medicine, Seattle, Washington, 98105, USA. 168
University of Washington
School
169
OXYGEN DISSOCIATION CURVE OF THE MONKEY
a\0 ’ 60 _$ cl : 50 9 3 40
30
20
IOI-
L
IO
20
30
40 50 02 Pressure-mmHg
60
70
80
3
Fig.1.Averageoxygendissociation curveof blood from 6 rhesus monkeys, showing individually measured points. The curve was constructed at a temperature of 38 “C and a pH of 7.40.
both of which affect 0, affinity (ROOTH and CALIGARA, 1961; ROOTH, SOMMERKAMP and BARTELS, 1962; KENNEDY and VALTIS, 1954). This curve is considerably to the right of that reported by BEHRMAN, HELLER, BATTAGLIA and HELLEGERS (1963) for blood from pregnant rhesus monkeys in Puerto Rico. These workers reported a T,, of 27.0 mm Hg at 38 “C and pH 7.40. The reason for the discrepancy is not clear at present. It seems unlikely that pregnancy is the reason for the difference in 0, affinity since in humans the position of the 0, dissociation curve when expressed at a constant pH is similar in pregnant (HELLEGERS and SCHRUEFER, 1961) and nonpregnant subjects (BARTELS and HARMS, 1959).The present group of animals included a male, a female with a retained placenta, and four nonpregnant females. The weight range was 4.8-7.8 kg. Use of a Bohr effect factor of -0.5 instead of -0.6 (BEHRMAN et al., 1963) introduces negligible error, as the pH values of the tonometered blood in the present study were close to 7.40 and distributed on each side of it. Differences in hemoglobin type are associated with differences in O2 affinity in
170
J. T. PARER
TABLE Gas pressures
and contents,
Animal No.
176
2278
2281
102
426
163
oxygen
I saturation
and pH of blood
Total
Total
Po, mm Hg
Pcoz mm Hg
co, vol 0%
cco, vol 0%
HbOz
207.0
45.0
17.9
45.6
100
59.1
46.6
14.2
47.0
81.0
7.320
44.1
46.7
11.2
48.6
64.2
7.320
26.3
47.4
5.7
50.0
32.3
7.330
14.4
46.2
2.3
50.8
12.8
7.350
216.0
48.0
15.1
47.0
71.1
44.9
12.7
46.8
86.3
58.1
45.8
II.5
46.6
78.8
7.320
45.0
47.6
9.0
48.4
61.9
7.305
41.1
49.7
8.2
49.4
55.6
7.300
31.1
46.3
6.6
49.4
45.1
7.323
13.2
46.3
1.5
50.8
9.8
7.335
216.0
48.0
15.6
58.2
71.1
45.0
13.6
56.8
90.0
58.3
45.8
12.9
57.0
85.2
7.415
45.0
47.6
10.5
59.8
69.8
7.395
PHI
%
100
100
7.300 7.315
7.395 7.405
41.2
49.8
9.9
61.0
65.7
7.400
31.2
46.3
6.9
60.0
45.6
7.420
26.7
48.2
6.1
60.8
40.2
7.415
13.4
46.3
I .4
61.6
9.1
7.420
216.0
47.8
18.8
55.1
71.0
44.8
16.6
54.9
100
7.380
90.6
7.420
44.9
47.5
13.8
56.9
75.6
7.395
41.1
49.6
12.8
58.3
70.2
7.400
38.7
45.0
12.3
54.3
67.4
7.420
31.1
46.2
9.2
56.9
50.3
7.420
26.7
48.1
7.0
58.9
41.9
7.410
14.6
46.8
2.5
60.3
13.8
7.425
214.0
47.7
18.0
50.3
44.7
16.3
48.5
100 93.4
7.434
70.7 57.9
45.6
14.9
49.7
85.1
7.375 7.372
44.6
47.3
12.0
52.1
68.4
7.360
40.9
II.5
52.3
66. I
7.355
30.9
49.5 46.1 46.
44.5 13.3
7.375
13.0
I
7.8
52.5
2.3
53.9
7.395
100
214.0
47.7 47.3
18.3 11.7
46.5
44.6 30.9
48.1
66.1
7.340
46.1
8.0
48.3
44.9
7.355
171
OXYGENDISSOCIATIONCURVE OF THE MONKEY
other species, e.g., sheep (HUISMAN, van VLIET and SEBENS, 1958; NAUGHTON et al., 1963; DAWSON and EVANS, 1965), but there was no suggestion of two populations of O2 dissociation curves in either group of monkeys. Hemoglobin of only one electrophoretic mobility was found in the Puerto Rican monkeys with techniques known to separate human hemoglobin types (BEHRMAN and SCHRUEFER, personal communication). TUTTLE et al. (1961) reported only one electrophoretic component in M. mulatta, though BUETTNER-JANUSCH et al. (1961) were able to separate up to 5 components in the same species. No 0, affinity measurements were reported by these workers. Fresh blood from two of the monkeys in this study was subjected to complete hemolysis (by severe bubbling of gas) with a resultant shift of the 0, dissociation curve approximately 5 mm to the left of that of the whole blood at 50 % saturation expressed at a pH of 7.40. The position of the curve of hemolyzed blood was almost identical with that reported by BEHRMAN et al. (1963) for whole blood in their monkeys. Computer analysis of the data obtained in the present study with the same program used by BEHRMAN et al. (1963) to calculate their 0, dissociation curves yielded the following relationship between Po2, pH and percent saturation (S): log Po, = 6.0718-0.6197
pH+0.3792
log
The T,, calculated from this is 31.0 mm Hg, i.e. 1.3 mm less than that obtained by hand-drawing curves and averaging the T,,. The use of the previously published 0, dissociation curve (BEHRMAN et al., 1963) will result in considerable error if applied to rhesus monkey blood with an 0, affinity similar to that reported here. It is suggested that direct measurements of both 0, tension and percent saturation be made in rhesus monkey blood until the discrepancy is clarified. Acknowledgements The author
is grateful
to Mr. R. Coffin and Dr. R. E. Behrman
for the use of their
program and assistance with the computer analyses and to Dr. James Metcalfe of the University of Oregon Medical School for encouragement and the use of facilities. This work is publication No. 142 from the Oregon Regional Primate Research Center and was supported by PHS training grant No. 5T15499, research grant No. HE 06042 from the National Heart Institute, and grant No. FR 00163 of the National Institute of Health.
BARTELS, H. P//tigers
and H. HARMS (1959). Arch.
Ges. Physiol.
Sauerstoffdissoziationskurven
des Blutes von Slugetieren.
268: 334-365.
BEHRMAN,R. E., C. J. HELLER, F. C. BAITAGLIA and A. E. HELLECERS(1963). A comparison oxygen affinity of maternal and fetal blood of the Macaca 258-264.
mulatta.
of the
Q. /. Expfl. Phvsiol.
48:
J. T. PARER
172
BUETTNER-JANUSCH, J., J. B. TWICHELL, B. Y-S. WONG and G. VAN WAGENEN (1961). Multiple haemoglobins
and transferrins in a macaque sibship. Nature 192 : 948-950.
DAWSON, T. J. and J. V. EVANS (1965). Effect of hemoglobin
type on the cardiorespiratory
system cf
sheep. Am. J. Physiol. 209: 593-598. HELLEGERS,A. and J. J. P. SCHRUEFER(1961). Nomograms
and empirical equations relating oxygen
tension, percentage saturation, and pH in maternal and fetal blood. Am. J. Ohsret. Gynec. 81:
377-384. HUISMAN,T. H. J., G. van VLIET.and T. SEBENS (1958). Sheep haemoglobins. KENNEDY,A. C. and D. J. VAL~IS (1954). The oxygen dissociation
Nature 182: 171-172.
curve in anemia of various types.
J. Clin. Invest. 33: 1372-1381. NAUGHTON, M. A., G. MESCHIA, F. C. BATTAGLIA,A. HELLEGERS,H. HOGOPIANand D. H. BARRON (1963). Hemoglobin
characteristics
and the oxygen affinity of the bloods
of Dorset
sheep.
Q. J. Exptl. Physiol. 48: 313-323. PARER,J. T., A. S. HOVERSLANDand J. METCALFE(1967). Some respiratory characteristics of the blood of the adult and young African pygmy goat. J. Appf. Physiol. (in press). ROOTH, G. and F. CALIGARA (1961). The influence of metabolic acid base variation on the oxygen dissocation
curve. Clin. Sci. 21: 393-401.
ROOTH, G., H. SOMMERKAMPand H. BARTELS(1962). The influence of base excess and cation concentration in the red cells on the position of the oxygen dissociation
curve. Clin. Sci. 23: 14.
TUTTLE, A. H., F. E. NEWSOME,C. H. JACKSONand R. R. OVERMAN(1961). Hemoglobin Macaca irus and Macaca mulatta monkeys. Science 133 : 578-579.
types of
Publishing Company, Amsterdam
THE O2 DISSOCIA~ON CURVE OF BLOOD OF THE RHESUS MONKEY (MACACA MULATTA)
J. T.
PARERI
Oregon Regionaf Primate Research Center, Beaverton, Oregon, and Heart Research Laboratory, University of Oregon Medical School, Portland, Oregon, USA
Abstract. The 0~ dissociationcurve of blood of rhesus monkeys was constructed
using blood samples from six animals. The mean 02 tension necessary for SOT! saturation of hemoglobin (Tso) was 32.3 mm Hg at 38 “C and plasma pH of 7.40. This is considerably to the right of the previously published curve for this species. Monkey blood Oxygen affinity of blood Oxygen dissociation curve
Materials and methods Oxygen dissociation curves were constructed with venous blood drawn from six rhesus monkeys into syringes containing fluoride in heparin. No hemolysis was evident. The monkeys were anesthetized with 0.3 to 3.0 mg/kg of phencyclidine hydrocholoride IM (Sernylan, Parke Davis and Co., Ann Arbor, Michigan). Aliquots of blood were tonometered at a temperature of 38 “C with gases of known 02, CO, and N, composition (analysed by the method of Scholander). OZ concentration of the blood was then measured by the method of Van Slyke and Neill, and pH was measured with a Radiometer glass electrode at 38 “C. The technique has been described more fully elsewhere (PARER, HOVERSLAND and METCALFE, 1967). Results and discussion Fig. 1 shows the average 0, dissociation curve, together with the measured points converted by means of a Bohr effect factor of -0.5 units to represent their values at pH 7.40. The measured and calculated data from which the curves were constructed are given in table 1. The CO, tensions and contents are also shown. The average O2 tension at 50% saturation (T,,) ranged from 31-33 mm Hg. The mean 0, capacity was 16.6 vol %; none of the subjects was in an abnormal acid-base status or anemic, Accepted for pablicat~on 21 November 1966. 1 Present Address: Department of Obstetrics and Gynecology, of Medicine, Seattle, Washington, 98105, USA. 168
University of Washington
School
169
OXYGEN DISSOCIATION CURVE OF THE MONKEY
a\0 ’ 60 _$ cl : 50 9 3 40
30
20
IOI-
L
IO
20
30
40 50 02 Pressure-mmHg
60
70
80
3
Fig.1.Averageoxygendissociation curveof blood from 6 rhesus monkeys, showing individually measured points. The curve was constructed at a temperature of 38 “C and a pH of 7.40.
both of which affect 0, affinity (ROOTH and CALIGARA, 1961; ROOTH, SOMMERKAMP and BARTELS, 1962; KENNEDY and VALTIS, 1954). This curve is considerably to the right of that reported by BEHRMAN, HELLER, BATTAGLIA and HELLEGERS (1963) for blood from pregnant rhesus monkeys in Puerto Rico. These workers reported a T,, of 27.0 mm Hg at 38 “C and pH 7.40. The reason for the discrepancy is not clear at present. It seems unlikely that pregnancy is the reason for the difference in 0, affinity since in humans the position of the 0, dissociation curve when expressed at a constant pH is similar in pregnant (HELLEGERS and SCHRUEFER, 1961) and nonpregnant subjects (BARTELS and HARMS, 1959).The present group of animals included a male, a female with a retained placenta, and four nonpregnant females. The weight range was 4.8-7.8 kg. Use of a Bohr effect factor of -0.5 instead of -0.6 (BEHRMAN et al., 1963) introduces negligible error, as the pH values of the tonometered blood in the present study were close to 7.40 and distributed on each side of it. Differences in hemoglobin type are associated with differences in O2 affinity in
170
J. T. PARER
TABLE Gas pressures
and contents,
Animal No.
176
2278
2281
102
426
163
oxygen
I saturation
and pH of blood
Total
Total
Po, mm Hg
Pcoz mm Hg
co, vol 0%
cco, vol 0%
HbOz
207.0
45.0
17.9
45.6
100
59.1
46.6
14.2
47.0
81.0
7.320
44.1
46.7
11.2
48.6
64.2
7.320
26.3
47.4
5.7
50.0
32.3
7.330
14.4
46.2
2.3
50.8
12.8
7.350
216.0
48.0
15.1
47.0
71.1
44.9
12.7
46.8
86.3
58.1
45.8
II.5
46.6
78.8
7.320
45.0
47.6
9.0
48.4
61.9
7.305
41.1
49.7
8.2
49.4
55.6
7.300
31.1
46.3
6.6
49.4
45.1
7.323
13.2
46.3
1.5
50.8
9.8
7.335
216.0
48.0
15.6
58.2
71.1
45.0
13.6
56.8
90.0
58.3
45.8
12.9
57.0
85.2
7.415
45.0
47.6
10.5
59.8
69.8
7.395
PHI
%
100
100
7.300 7.315
7.395 7.405
41.2
49.8
9.9
61.0
65.7
7.400
31.2
46.3
6.9
60.0
45.6
7.420
26.7
48.2
6.1
60.8
40.2
7.415
13.4
46.3
I .4
61.6
9.1
7.420
216.0
47.8
18.8
55.1
71.0
44.8
16.6
54.9
100
7.380
90.6
7.420
44.9
47.5
13.8
56.9
75.6
7.395
41.1
49.6
12.8
58.3
70.2
7.400
38.7
45.0
12.3
54.3
67.4
7.420
31.1
46.2
9.2
56.9
50.3
7.420
26.7
48.1
7.0
58.9
41.9
7.410
14.6
46.8
2.5
60.3
13.8
7.425
214.0
47.7
18.0
50.3
44.7
16.3
48.5
100 93.4
7.434
70.7 57.9
45.6
14.9
49.7
85.1
7.375 7.372
44.6
47.3
12.0
52.1
68.4
7.360
40.9
II.5
52.3
66. I
7.355
30.9
49.5 46.1 46.
44.5 13.3
7.375
13.0
I
7.8
52.5
2.3
53.9
7.395
100
214.0
47.7 47.3
18.3 11.7
46.5
44.6 30.9
48.1
66.1
7.340
46.1
8.0
48.3
44.9
7.355
171
OXYGENDISSOCIATIONCURVE OF THE MONKEY
other species, e.g., sheep (HUISMAN, van VLIET and SEBENS, 1958; NAUGHTON et al., 1963; DAWSON and EVANS, 1965), but there was no suggestion of two populations of O2 dissociation curves in either group of monkeys. Hemoglobin of only one electrophoretic mobility was found in the Puerto Rican monkeys with techniques known to separate human hemoglobin types (BEHRMAN and SCHRUEFER, personal communication). TUTTLE et al. (1961) reported only one electrophoretic component in M. mulatta, though BUETTNER-JANUSCH et al. (1961) were able to separate up to 5 components in the same species. No 0, affinity measurements were reported by these workers. Fresh blood from two of the monkeys in this study was subjected to complete hemolysis (by severe bubbling of gas) with a resultant shift of the 0, dissociation curve approximately 5 mm to the left of that of the whole blood at 50 % saturation expressed at a pH of 7.40. The position of the curve of hemolyzed blood was almost identical with that reported by BEHRMAN et al. (1963) for whole blood in their monkeys. Computer analysis of the data obtained in the present study with the same program used by BEHRMAN et al. (1963) to calculate their 0, dissociation curves yielded the following relationship between Po2, pH and percent saturation (S): log Po, = 6.0718-0.6197
pH+0.3792
log
The T,, calculated from this is 31.0 mm Hg, i.e. 1.3 mm less than that obtained by hand-drawing curves and averaging the T,,. The use of the previously published 0, dissociation curve (BEHRMAN et al., 1963) will result in considerable error if applied to rhesus monkey blood with an 0, affinity similar to that reported here. It is suggested that direct measurements of both 0, tension and percent saturation be made in rhesus monkey blood until the discrepancy is clarified. Acknowledgements The author
is grateful
to Mr. R. Coffin and Dr. R. E. Behrman
for the use of their
program and assistance with the computer analyses and to Dr. James Metcalfe of the University of Oregon Medical School for encouragement and the use of facilities. This work is publication No. 142 from the Oregon Regional Primate Research Center and was supported by PHS training grant No. 5T15499, research grant No. HE 06042 from the National Heart Institute, and grant No. FR 00163 of the National Institute of Health.
BARTELS, H. P//tigers
and H. HARMS (1959). Arch.
Ges. Physiol.
Sauerstoffdissoziationskurven
des Blutes von Slugetieren.
268: 334-365.
BEHRMAN,R. E., C. J. HELLER, F. C. BAITAGLIA and A. E. HELLECERS(1963). A comparison oxygen affinity of maternal and fetal blood of the Macaca 258-264.
mulatta.
of the
Q. /. Expfl. Phvsiol.
48:
J. T. PARER
172
BUETTNER-JANUSCH, J., J. B. TWICHELL, B. Y-S. WONG and G. VAN WAGENEN (1961). Multiple haemoglobins
and transferrins in a macaque sibship. Nature 192 : 948-950.
DAWSON, T. J. and J. V. EVANS (1965). Effect of hemoglobin
type on the cardiorespiratory
system cf
sheep. Am. J. Physiol. 209: 593-598. HELLEGERS,A. and J. J. P. SCHRUEFER(1961). Nomograms
and empirical equations relating oxygen
tension, percentage saturation, and pH in maternal and fetal blood. Am. J. Ohsret. Gynec. 81:
377-384. HUISMAN,T. H. J., G. van VLIET.and T. SEBENS (1958). Sheep haemoglobins. KENNEDY,A. C. and D. J. VAL~IS (1954). The oxygen dissociation
Nature 182: 171-172.
curve in anemia of various types.
J. Clin. Invest. 33: 1372-1381. NAUGHTON, M. A., G. MESCHIA, F. C. BATTAGLIA,A. HELLEGERS,H. HOGOPIANand D. H. BARRON (1963). Hemoglobin
characteristics
and the oxygen affinity of the bloods
of Dorset
sheep.
Q. J. Exptl. Physiol. 48: 313-323. PARER,J. T., A. S. HOVERSLANDand J. METCALFE(1967). Some respiratory characteristics of the blood of the adult and young African pygmy goat. J. Appf. Physiol. (in press). ROOTH, G. and F. CALIGARA (1961). The influence of metabolic acid base variation on the oxygen dissocation
curve. Clin. Sci. 21: 393-401.
ROOTH, G., H. SOMMERKAMPand H. BARTELS(1962). The influence of base excess and cation concentration in the red cells on the position of the oxygen dissociation
curve. Clin. Sci. 23: 14.
TUTTLE, A. H., F. E. NEWSOME,C. H. JACKSONand R. R. OVERMAN(1961). Hemoglobin Macaca irus and Macaca mulatta monkeys. Science 133 : 578-579.
types of