/. Embryol. exp. Morph. Vol. 25, 1, pp. 33-45, 1971
Printed in Great Britain
33
Ontogeny of hemoglobin in the house sparrow
By FRANCIS M. BUSH* AND J. 1YES TOWNSEND 2
From the Department of Anatomy and Department of Genetics,
Medical College of Virginia
SUMMARY
A 'switchover' from synthesis of predominantly embryonic hemoglobin to synthesis of
predominantly adult hemoglobin occurs during differentiation of the house sparrow. This
'switchover' is demonstrated by a change in electrophoretic pattern generally comparable
with that of the chicken, slightly different from those of the duck and man, and, perhaps,
dissimilar to that of the red-wing blackbird.
Differences in electrophoretic mobility show that 10-day embryos possess one hemoglobin
not present in early hatchlings and two hemoglobins not present in adults. Major adult
hemoglobin is synthesized by the tenth embryonic day; and minor adult hemoglobin, between
the tenth embryonic day and the seventh day after hatching.
The major increase in concentration of hemoglobin occurs between 2 and 11 days after
hatching, a time when the two predominant embryonic and two predominant adult hemoglobins are being synthesized. The percentage of packed cells (hematocrit) also increases significantly within this period.
Similarities of peptide maps for the house sparrow and other species, such as the owl, indicate the presence of a-and/?-polypeptide chains in adult house sparrow hemoglobin. The shift
to hemoglobins with faster anodic mobilities and the presence of a higher concentration of
hemoglobin in adults than in early hatchlings suggest that this synthesis is predominantly in
the /?-chains.
INTRODUCTION
Changes in hemoglobin patterns during morphogenesis are documented for
some species of birds. Early embryonic hemoglobins, distinct from hemoglobins
of late embryos (fetuses), hatchlings, and adults, exist in a number of precocial
species, such as turkey, Meleagris gallopavo, and chuckar partridge, Alectoris
graeca (reviewed by Man well, Baker & Betz, 1966), chicken, Gallus gallus
(Manwell et al. 1966; Godet, 1967; Atherton, 1969), and white Pekin duck,
Anas domesticus (Borgese & Bertles, 1965). Little is known of the ontogeny of
hemoglobins in small altricial species; the only published study is limited to an
analysis of embryonic stages in the red-wing blackbird, Agelaiusphoeniceus. In
this species, no time-specific alteration occurs during embryonic stages, for the
hemoglobin patterns of 2-day and 14-day embryos are identical (Manwell,
Baker, Roslansky & Foght, 1963).
1
Author's address: Department of Anatomy, Health Sciences Division, Virginia Commonwealth University, Richmond, Virginia, 23219, U.S.A.
2
Author's address: Department of Genetics, Health Sciences Division, Virginia Commonwealth University, Richmond, Virginia, 23219, U.S.A.
3
EM B 25
34
F. M. BUSH AND J. I. TOWNSEND
Time-specific changes vary for different precocial species. The appearance of
some new adult hemoglobins occurs by the sixth day in chicken (Hashimoto &
Wilt, 1966; Man well et al. 1966; Godet, 1967), by the sixth, seventh and eighth
days, in chuckar partridge, bantam chicken and turkey, respectively (Manwell
et al. 1966), and by the eighth day in Pekin duck (Borgese & Bertles, 1965). The
' switchover' from embryonic to hatchling or to adult hemoglobins is not entirely
perfect because synthesis of the new component can occur before synthesis of
the embryonic component ceases. Ontogenetic changes show that some hemoglobins formed early are transient and others are newly synthesized during either
fetal or hatchling stages, or during both. Starch gel patterns reveal that an acidic
component and a basic component of chick embryos appear by the seventh and
tenth days, respectively, while another component, a major acidic hemoglobin,
disappears by the ninth embryonic day (Godet, 1967). The hemoglobin appearing by the seventh day persists in the adult, and that appearing on the tenth day
disappears during the first month after hatching; on the other hand two other
acidic components develop by the fourth day after hatching, and both persist in
the adult. The 7-day embryonic Pekin duck forms one acidic component that
disappears by the tenth week after hatching (Borgese & Bertles, 1965).
The observed differences between precocial and altricial species and the
paucity of information about small species of feral birds prompted the present
study, one of the continuing series (Bush, 1967; Bush & Seibert, 1968; Bush, Price
& Townsend, 1970) to discover the biochemical properties of blood of the
altricial species, house sparrow, Passer domesticus. This paper reports, correlates,
and discusses in relation to what is known of other species, the quantity, composition and development of major and minor hemoglobins in embryo, hatchling
and adult; the concentration of hemoglobin and measurement of hematocrit in
hatchling and adult; and the mapping of peptides in the adult.
MATERIALS AND METHODS
The sample. The house sparrows, Passer domesticus, in our study were collected
from the population at Richmond, Virginia. In all, 53 adult females and 69 adult
males (older than 1 year), 127 hatchlings and juveniles (less than 50 days old),
60 first-year adults (50-124 days old), and 10 embryos (10-12 days old) were
studied. Specimens were assigned relative ages based on criteria described
elsewhere (Farrar & Bush, 1969).
Collection of blood and determination of hematocrit. Blood was obtained by
capillary pipette after embryonic vitelline vessels were cut, and by syringe from
the hearts of hatchlings and adults. The samples were heparinized. Duplicate
samples of hatchling and adult blood were centrifuged at 14000g for 10 min in a
microhematocrit centrifuge (Clay-Adams), and the mean hematocrit value
determined.
Quantitation of hemoglobin. The modified Wong technique (MacFate, 1964)
Ontogeny of hemoglobin
35
was used to determine the iron content in hemoglobin. This technique involves
detachment of iron from the hemoglobin molecule by treatment with sulfuric
acid and potassium persulfate, precipitation of proteins with sodium tungstate,
and treatment of the supernatant fluid with potassium thiocyanate. Light
transmission at 540 m/t by the supernatant fluid was then measured in a Bausch
and Lomb Spectronic '20' colorimeter at 25 °C. Each per cent transmittance
value was compared with per cent transmittance values obtained from a calibration curve prepared by serially diluting a stock solution of ferrous iron
(0-1 mg iron/ml). For each blood sample, the concentration was:
g hemoglobin =
mg iron/100 ml
3-4
Preparation of hemoglobin. After centrifugation of blood samples at 2000^
for 15 min, plasmas were drawn off. The cells remaining in the centrifuge tubes
were washed three times with 1 % saline solution and then exposed to carbon
monoxide to form the CO-hemoglobin derivative. Cells were hemolyzed in
four volumes of distilled water and one volume of toluene, and then rotated for
2 h at 4 °C. This mixture was centrifuged at lOOOOg for 1 h, and the clear red
hemoglobin layer was filtered to remove any traces of cell debris.
Starch gel electrophoresis. Samples were subjected to vertical starch gel
electrophoresis (Smithies, 1959). Each gel slot contained approximately the
same amount of hemoglobin solution. Gels were prepared with a Tris-EDTAborate buffer, pH 8-6 (Manwell et al. 1966). Electrophoresis was for 7 h at 4 °C,
with power of 280 V, supplied by a Duostat (Beckman). Gels were split longitudinally, and one-half was stained for hemoglobin by a standard peroxidative
procedure using benzidine and hydrogen peroxide in acetate buffer, pH 4-7
(Dessauer, 1966). Reactions were retarded after 5 min by washing gels with
0-5% saline solution. The other half was stained with amido black and
cleared; then both halves were photographed. Quantitative measurement of the
predominant hemoglobins was determined by transmission densitometry of the
photographic negative of starch gels stained with amido black by the technique
of Hecht, Motulsky, Lemire & Shepard (1966). Negatives were scanned with
an automated broad-range microspectrophotodensitometer as described by
Williams, Ruffin & Berry, (1964).
Fingerprinting. Peptide mapping was done according to the method of Murayama (1964). Samples of purified hemoglobins were heat denatured and digested
with trypsin; the pH of the sample was adjusted to 6-5 with 1 N-HC1; the precipitate was removed by centrifugation; and then the samples were lyophilized.
Samples of hydrolysates dissolved in ethanol were spotted on Whatman 3 MM
filter paper sheets and electrophoresed with a water-acetate-pyridine (287:10:1)
buffer, pH 3-7, at 1100 V for 2 h, on a flat plate electrophoretic unit (Savant).
At right angles to the axis of electrophoresis, the sheets were subjected overnight to ascending chromatography with a solvent system of water-butanol3-2
36
F. M. BUSH AND J. I. TOWNSEND
acetate (5:4:1). After the paper was dried, peptides were localized by being
stained with 0-25 % ninhydrin in ethanol.
RESULTS
Quantitation of hemoglobin concentration shows that there is increasing
synthesis after hatching (Fig. 1). Hatchlings of 2-5 days have approximately
one-fourth the hemoglobin content of adults. The greatest synthesis, approximately one and one-half fold, occurs from 2 to 11 days after hatching.
Although the hemoglobin content increases in each succeeding stage of development, the increase from the tenth day is slightly more than one-half fold. The
mean concentration of hemoglobin of males is slightly, but not significantly
(for /-test, P > 0-05), higher than that of females.
24
200
19
E
o 150
I
27
-r
31
J
J I
JL
f I
UjJ
LjJ
-E, 100
o
E
50
2-5H
6-7
8-10 11-14 15-30 31-49 50-80 81-124
Days
Fig. 1. Concentration of hemoglobin at different developmental stages. Central horizontal line = mean; stippled area = 1 S.D.; vertical line = range; and open area =
2 S.E. The number of individuals sampled in each age group is at the top of each
vertical line.
The measurement of hematocrit reveals that the percentage of packed cells
steadily increases after hatching (Fig. 2). Newly hatched young have less than
one-half of the percentage of packed cells present in the adult. The mean value
for packed cells of males is also slightly, but not significantly (for /-test,
P > 005), higher than that of females.
Typical starch gel electrophoretic resolution of embryo, hatchling, and adult
hemoglobins stained by the peroxidative procedure and by amido black is
shown in Fig. 3. These results show that there is a developmental change, and
that the synthesis of adult hemoglobin occurs by at least the tenth embryonic
day. Designation of predominant hemoglobins follows the same designation
Ontogeny of hemoglobin
37
as applied for predominant hemoglobins of other avian species undergoing
differentiation. Predominant zones of the three patterns are: (1) Ten-day embryos
have an acidic major embryonic hemoglobin which resolves into two components;
the more acidic of the two comprises over one-fourth of the total hemoglobin
(Table 1). The most basic minor embryonic hemoglobin makes up slightly less
than one-fifth of the total. These embryo patterns lack the most acidic compo-
600
69
53
450 -
24
7
10
300
31
n
12
150 -
2-5H
6-10 11-14
15-30 31-49 50-:80 81-124
Days
Fig. 2. The mean hematocrit at different developmental stages. The number of
individuals sampled in each age group is at the top of each bar.
Adult
minor Hb
••i
Embryo
major Hb
Adult
major Hb
Embryo
minor Hb
Adult
7-day
10-day
hatchling
embryo
Adult
7-day
10-day
hatchling
embryo
B
Fig. 3. Electrophoretic patterns of predominant hemoglobins of an adult, a 7-day
hatchling, and a 10-day embryo, stained with benzidine (A) and the comparable patterns stained with amido black (B).
38
F. M. BUSH AND J. I. TOWNSEND
nent found in early hatchling and adult patterns. (2) Seven-day hatchling
patterns retain the minor embryonic hemoglobin, but they also have the minor
adult hemoglobin. The percentage of major and minor embryonic hemoglobins
decreases after hatching, while the percentages of the major adult hemoglobin
and of the minor adult hemoglobin increase during the same period (Table 1).
These two adult hemoglobins constitute almost one-half of the total quantity
in the 7-day hatchling pattern. (3) Adult house sparrow patterns have at least
two predominant zones. The major adult hemoglobin comprises nearly one-third
of the total quantity, and the minor adult hemoglobin makes up about one-fifth of
the total (Table 1). The major embryonic hemoglobin is still synthesized, but the
minor embryonic hemoglobin is not.
Table 1. Relative proportions of predominant hemoglobins in
embryo, hatchling and adult
Hemoglobin (%)*
A
Age
10-day embryo
7-day hatchling
Adult
Adult
major
Adult
minor
Embryo
major
Embryo
minor
Other
23
27
29
0
20
19
28
19
18
18
14
0
31
20
34
* Transmission densitometry of negative of starch gel stained with amido black.
There are other hemoglobins present in these individuals of different ages
(Table 1). Densitometric tracings confirm that as many as seven different hemoglobins are synthesized during development. The tracings reveal three peaks other
than the major and minor embryonic and adult components. One of these is the
slower acidic component near the gel insertion slot, and the other two are basic
components. One basic component migrates near the slot; the other basic
component is prominent and migrates to a position intermediate between the
major adult and the minor embryonic hemoglobins. The slower acidic component of the major embryonic hemoglobin is indistinguishable from the major
embryonic hemoglobin on tracings of 7-day hatchling and adult patterns.
Electrophoretic patterns of female house sparrow hemoglobins resemble those
of males.
Comparison of the electrophoretic patterns for the developmental stages of
the house sparrow shows that the time of switching off or on synthesis of the
different hemoglobins varies considerably: for example, while synthesis of both
major hemoglobins apparently begins in the embryo and continues throughout
all stages of the life cycle, synthesis of the minor adult hemoglobin is switched
on near the time of hatching, about the time when synthesis of the trace type of
embryonic hemoglobin, which migrates between the major adult and minor
embryonic hemoglobin, is switched off.
Ontogeny of hemoglobin
39
The characteristic fingerprint of purified adult house sparrow hemoglobin is
shown in Fig. 4. The pattern shows the presence of 19 peptides. Four of these
peptides stain lightly with ninhydrin, and 15 stain intensely.
Fig. 4. Schematic composite of tryptic peptide map of adult hemoglobin. Highvoltage electrophoresis is in the horizontal dimension and ascending chromatography,
in the vertical dimension. Arrow indicates point of application of tryptic digest.
Stippling indicates density of staining with ninhydrin.
DISCUSSION
Study of different stages of the life cycle of the house sparrow shows an orderly
sequence of synthesis of different hemoglobins. This is evident in spite of technical problems that have so far prevented study in the earliest embryonic
stages when the amount of hemoglobin present is so limited that purification is
difficult, a difficulty that may soon be overcome by application of microelectrophoretic techniques, such as applied by Matioli & Niewisch (1965) to separate
the hemoglobin of a single erythrocyte.
The sequence of synthesis of hemoglobins in the house sparrow differs from
that in its relative, the red-wing blackbird, which also belongs to the Order
Passeriformes. In the red-wing blackbird, all component hemoglobins persist
throughout the 14 days of embryonic development, that is, no 'switch over' to a
uniquely adult pattern occurs during late embryonic development. Manwell et
al. (1963) note similarities between this persistence of embryonic hemoglobins
and the persistence of fetal hemoglobins in adult human beings who have the
'high Hb F ' trait, which is presumably caused by a mutation in an operator
gene (Neel, 1961; Motulsky, 1962). Such a comparison, although interesting,
40
F. M. BUSH AND J. I. TOWNSEND
may be premature, for, as Manwell et al. (1963) note, hemoglobins of the adult
red-wing blackbird have not been studied and it is not known whether the
embryonic components persist through the hatchling stages. There is variation
between the different hemoglobins of the house sparrow in the time at which
their synthesis is switched on or off. Discovery of some such variation in the
red-wing blackbird might be found if hatchling and adult patterns are studied.
However, the difference in the time sequence in the two related species could be
caused by a different (mutated) operator gene or by the same operator gene
acting in different genetic backgrounds; no evidence upon which to base a
choice between these possibilities is available.
The separation of adult sparrow hemoglobin into five bands shows that
greater resolution is obtained by starch gel electrophoresis than is obtained
by paper electrophoresis. Using paper as the supporting medium, Ghosh
(1965) separated two anodally migrating hemoglobin bands in this species; the
faster makes up 72 % of the total hemoglobin, and the slower 28 %. Our greater
separation of hemoglobin in the adult house sparrow shows that the number of
hemoglobin bands in this species is comparable with that found by starch gel
electrophoresis of chicken hemoglobins (Manwell et al. 1966; Godet, 1967).
However, there are species differences in the number of hemoglobin bands. The
number for the house sparrow is greater than that found either in the large
altricial species of great horned owl, Bubo virginianus, or in the duck. In the
adult owl, a single anodally migrating band occurs on either filter paper, starch
gel or agar gel electropherograms (Abercrombie, Maber & Vella, 1969). In the
duck, only two bands separate by acrylamide gel electrophoresis (Borgese &
Bertles, 1965) and two fractions by ion-exchange chromatography (Klein,
1970).
Hemoglobin synthesis in birds is heterogeneous. The minor adult hemoglobin
appears later in the development of the house sparrow than does the major
adult hemoglobin. Photographs and diagrams of gels show a similar pattern of
synthesis in the chicken (Manwell etal. 1963; Manwell etal. 1966; Godet, 1967).
But the pattern is reversed in the duck where the major adult hemoglobin is
synthesized later in development than is the minor adult hemoglobin (Borgese &
Bertles, 1965; Klein, 1970). The discovery of a single hemoglobin band on
electropherograms of the adult great horned owl (Abercrombie et al. 1969)
suggests a simpler pattern of synthesis than in these other species. The apparent
differences beween the patterns of hemoglobin synthesis in these birds, as well as
the absence of a detectable 'switchover' in hemoglobin in the red-wing blackbird embryo (Manwell et al. 1963), make a further study of patterns of hemoglobin synthesis in avian species interesting and important.
A major increase in hemoglobin concentration and inhematocrit accompanies
the shift in electrophoretic properties of hemoglobins after hatching in the house
sparrow. The increase in hemoglobin concentration is greater than onefold and
in hematocrit almost onefold during 2-11 days after hatching. These rapid
Ontogeny of hemoglobin
41
increases result from the simultaneous synthesis of embryonic and adult
hemoglobins.
Concentrations of hemoglobin found during the early hatchling period of
house sparrows in the present study agree with the following values Kalabukhov
& Rodionov (1934) report in pooled samples of the house sparrow and European
tree sparrow, Passer montanus, hatchlings: 4-0 g/100 ml at 1-5 days, 6-8 at
10-14 days, 7-7 at 16-20 days, and 11-4 at 21-30 days. But the concentration in
42-day hatchling chickens, 10-1 g/100 ml (Lucas & Jamroz, 1961), is lower
than that, 13-6 g/100 ml, which we find in house sparrows of the same age.
The mean concentration of hemoglobin (17-0 g/100 ml) in our adult house
sparrows is somewhat greater than the following values reported (Nice, Nice &
Kraft, 1935) for presumably adult birds of seven other species also belonging to
the Order Passeriformes: 13-5 in tufted titmouse, Parus bicolor; 160, brown
thrasher, Toxostoma rufum.; 16-5, bronzed grackle, Quiscalus quiscala; 16-8,
cardinal, Pyrrhyloxia cardinalis; 15-8, towhee, Pipilo erythrophthalamus; 15-8,
junco, Junco hyemalis; and 13-4, white throated sparrow, Zonotrichia albicollis.
The value for the adult house sparrow is also greater than the 13-5 found
(Lucas & Jamroz, 1961) in the rooster, Callus gallus, Order Galliformes, and the
14-3 and 15-4 found (Riddle & Braucher, 1934) respectively in the dove (species ?)
and pigeon, Columba livia, Order Columbiformes; but it is approximately equal
to the 17-1 found (McFarland, 1963) in the western gull, Lams occidentalis,
Order Charadriiformes.
The slightly higher concentration of hemoglobin in the adult male house
sparrow (17-2 g/100 ml) than in the female (16-7) is consistent with sexual
differences found in the dove and the pigeon: male doves and male pigeons
have 14-6 and 160, respectively, and female doves and pigeons 140 and 14-7,
respectively (Riddle & Braucher, 1934).
The mean hematocrit in the house sparrow is greater than the reported value
for the corresponding stage of the chicken or western gull: 42-day hatchling—
chick 30-9% (Lucas & Jamroz, 1961), sparrow 40-5%; adult—rooster 40-0%
(Lucas & Jamroz, 1961), sparrow male, 51-6 %; and adult—gull 48-7 % (McFarland, 1963), sparrow 50-8%.
The differences in hemoglobin concentration and hematocrit reported between
house sparrows and other birds may be partially attributable to differences in the
techniques used and perhaps also to age differences, since often the ages of the
specimens sampled are not reported. We believe, however, that standardization
of techniques and ages would doubtlessly reveal species differences, although
perhaps of different magnitudes and even directions. These caveats by no means
denigrate the validity of the increase in hemoglobin concentration and hematocrit observed during development within the species.
While variations in hematocrit and concentration of hemoglobin occur during
the course of development of the house sparrow, variations in some other properties of hemoglobin, namely, alkali resistance, oxygen affinity and Bohr effect,
42
F. M. BUSH AND J. I. TOWNSEND
are known to occur during development of several vertebrate species. The hemoglobin of the embryonic chick has a higher oxygen affinity and exhibits a lower
Bohr effect than that of the adult chicken (Manwell et al. 1963). Hemoglobin of
newly hatched chicks and ducks is more alkali resistant than the adult hemoglobin of the same species; thus 13 % of the hemoglobin of newly hatched ducks
is denatured by alkali as is 27% of the hemoglobin of adults (Roberiro &
Villela, 1956). Adult house sparrow hemoglobin is less alkali resistant than is
adult chicken hemoglobin; some 66% is denatured in 1 min, and 92% of the
most anodally migrating fraction in 15 min (Ghosh, 1965). In man, fetal hemoglobin (a2y2) is resistant to alkali denaturation; in adult humans heterozygous
for Hb Rainier the hemoglobin has high oxygen affinity and is resistant to
alkali; and substitution of histidine in the /?-chain for H23 tyrosine prevents
denaturation (Stamatoyannopoulos, Yoshida, Adamson & Heinenberg, 1968).
The predominance of embryonic hemoglobin, low Bohr effect, and high oxygen
affinity are considered physiologically significant during some stages of development, for example in the chicken before vascularization of the egg surface and
air sac (Manwell et al. 1966). Presumably, in the early development of the house
sparrow, these same properties compensate for the low concentration of available hemoglobin and for the proportionately few erythrocytes.
Tryptic hydrolysates show that the hemoglobin of the adult house sparrow
has at least 19 peptides. This is in close agreement with the 20 peptides discovered for the Japanese quail, Coturnix coturnix (Manwell, 1963; Manwell et al.
1963), and also with the 23 found in the globin of the great horned owl (Abercrombie et al. 1969); but it is considerably fewer than the 33 peptides identified
in hemoglobin of the chicken and hemoglobin of the turkey (Manwell, 1963;
Manwell et al. 1963). In the great horned owl, the globin of the 3-week hatchling
has a- and ^-polypeptide chains, while the globin of the adult has a- and /?polypeptide chains (Abercrombie et al. 1969). The fingerprint pattern of the
adult house sparrow resembles the fingerprint pattern of the adult great horned
owl closely enough to suggest that the hemoglobin of the adult house sparrow
also has two different pairs of polypeptide chains. The electrophoretic shift from
predominantly basic to predominantly acidic hemoglobins in the developing
house sparrow implies a shift in predominant polypeptide chains. If, as in man,
cathodic migration means a predominance of a-chains (Huehns & Shooter,
1965), then these chains are predominant in embryonic house sparrow hemoglobin. The appearance of a different form of hemoglobin after hatching in the
house sparrow is similar to that of several species, including chicken (Manwell
et al. 1966), duck (Borgese & Bertles, 1965), great horned owl (Abercrombie et al.
1969), and man (Huehns & Shooter, 1965), where the hemoglobin synthesized
after hatching or following birth is predominantly of the /?-chain type.
It appears to us highly desirable to make further studies of hemoglobins in
house sparrows. Studies should be made of the electrophoretic properties,
alkali resistance, Bohr effect and oxygen equilibrium of hemoglobins, especially
Ontogeny of hemoglobin
43
in very early embryos; the globins should be isolated and their respective polypeptide chains identified; and the predominant peptides should be mapped.
These studies may disclose the presence either of earlier formed hemoglobins or
variant hemoglobins in the adult house sparrow population and disclose in this
species other developmental changes in hemoglobin similar to those in other
species. Localization of peptides and subsequent identification of their predominant amino acids would be expected to show differences in primary structure that contribute to the observed time-sequence for this species.
RESUME
Ontogenie de 1'hemoglobine chez le Moineau domestique
Pendant le developpement du moineau domestique on observe un passage de la synthese
predominate d'hemoglobine de type foetal a la synthese predominanted'hemoglobine adulte.
Ce passage est revele par un changement dans les electrophoregrammes; cechangement est
comparable a celui observe chez le poulet, semblable a celui observe chez le canard et chez
l'homme, mais semble different de celui observe chez le merle.
Des differences dans les mobilites electrophoretiques montrent que l'embryon de 10 jours
possede une hemoglobine qui n'est plus presente chez l'individu juste apres l'eclosion, et
deux hemoglobines qui sont absentes chez Padulte. De 1'hemoglobine adulte majeure est
synthetisee vers le lOeme jour de la vie embryonnaire; 1'hemoglobine adulte mineure, entre
le lOeme jour de la vie embryonnaire et vers le 7eme jour apres l'eclosion.
L'augmentation majeure de la concentration de 1'hemoglobine a lieu entre les 2eme et
I leme jours apres l'eclosion; a ce stade les deux hemoglobines embryonnaires predominates
et deux hemoglobines adultes predominates sont synthetisees. La valeur de Phematocrite
augmente egalement de facon significative pendant cette periode.
Des similarites entre les cartes peptidiques du moineau domestique et celles d'autres
especes, dont le hibou, indiquent la presence de chaines polypeptidiques alpha et beta dans
1'hemoglobine du moineau domestique adulte. Le passage a des hemoglobines migrant
plus rapidement vers l'anode, et la presence d'une concentration plus forte en hemoglobine
chez l'adulte que chez l'individu juste apres l'eclosion, suggerent que cette synthese concerne
essentiellement les chaines beta.
We are grateful to W. W. Farrar, Department of Biochemistry, Virginia Polytechnic
Institute and State University, for assistance with collection of these data, and to Dr Edward
S. Kline, Department of Biochemistry, Medical College of Virginia, for making the densitometric tracings.
This investigation was supported partly by NIH Grant GM 13649-02, to F. M. B.
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{Manuscript received 22 April 1970)