- Email: [email protected]
Some Aspects of 59Fe Distribution in Chicks and its Incorporation into Hemoglobin
Some Aspects of 59Fe Distribution in Chicks and its Incorporation into Hemoglobin J . GODET AND M . BELHAN1*
Section de Biologie Generate et Appliquee Laboratoire Associe au C.N.R.S., Universite Claude Bernard de Lyon, 43, boulevard du 11 novembre 1918 69621 Villeurbanne, France (Received for publication July 14, 1973)
POULTRY SCIENCE 53: 914-918, 1974
JN VIVO labelling of hemoglobin by 59Fe -t-involves all of the complex mechanisms of storage and exchange of this radioactive isotope in and between the various parts of the organism. These mechanisms are relatively unknown in poultry which have not reached the adult stage. Hence, to study avian hemoglobin synthesis during post-hatching life, it became necessary to define the main pathways of radioiron uptake in the developing chick. 59Fe was thus injected, in different ways, to 3 day old chicks and the subsequent labelling of various organs was followed for a week. Attention was focused mainly on the interactions, between the internal exchange of 59Fe and its incorporation into hemoglobin, which occured during that week. MATERIAL AND METHODS White Leghorn young chicks weighing 35-45 g. received by intraperitoneal or intravitelline injection 50 to 100 ux. of 59Fe (ferric citrate; average specific activity 12 mCi./mg. Fe). The exact amount of radioactivity injected was determined by weighing both the aliquot of the material used as a standard and the amount injected into the
'Present address: Centre Pierre et Marie Curie, Service d'Hematologie, Alger.
animal. At the time of sacrifice, the body weight of each animal was determined and a measured volume of blood was withdrawn from the wing vein. A first fraction of this blood was used to measure, as described elsewhere (Godet et al., 1970 a), blood hemoglobin content and hematocrit. A second fraction was used to measure the radioactivity of known aliquots of whole blood, plasma and red blood cells, after, in the case of red blood cells, five washes with NaCl 0.9%. The rest of the collected blood was used to prepare hemolysates from which hemoglobin was purified by gel filtration on Sephadex G-75 (Godet et al, 1970b). Once killed by decapitation, each animal was carefully dissected. Easily separated organs, like tibia, femur, spleen, liver, etc . . . were placed directly in counting tubes, the large organs being first cut in small pieces. The remaining carcass which includes skeleton, muscles, skin and feathers was first coarsely ground and then distributed in counting tubes. Radioactivity of the various organs, of blood and feces was measured in a Packard Automatic Gamma Counter with less than 5% variation. The results were expressed as percent of injected radioactivity and corrections were made, as far as organs are concerned, for the activity due to their blood content. The organ blood content was calculated from organ weight and total blood volume. The
914
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
ABSTRACT In order to study avian hemoglobin synthesis, 59Fe distribution and exchanges between various organs and erythrocyte components have been examined after injection in 3 day old chicks. Results obtained during the week after injection show that the overall ferrokinetics follow the same trends as those observed in most animals. The only differences noted can be attributed to the rapid growth of the chicks at that age.
59
915
F E DISTRIBUTION
TABLE 1.—Organ radioactivity after intraperitoneal injection of S9Fe
total blood volume was, in turn, calculated from the body weight (Godet et al., 1970a). RESULTS Animals, sacrificed at different times during the 24 hours which follow injection, show
that the amount of 59Fe recovered represents within ± 8% the amount of administered radioactivity (Table 1). After intraperitoneal injection, about 40% of the administered 59 Fe is found, four hours later, in a group of tissues formed by the
% o t injected radioactivity
40
20.
4h
12h
24 h
»«»...»„...//.*.... —' // 48h
•
" 6d time after injection
FIG. 1. 59Fe exchanges between blood, bone marrow, liver and digestive tract, over a week period after injection to 3 day old chicks
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
% of injected radioactivity 8 hours 4 hours 24 hours 11.90 4.46 \ } 26.35 5.45 / 9.90 14.45 5.*>">» 13.08 11.40 15.12 6.34 8.11 5.77 4.161 4.60 4.34) 4.00 > 9.05 1.94 4.17 > 9.52 6.80 0.89; 0.26 1.01 ) 0.06 0.38 0.04 0.61 0.73 0.59 0.12 0.06 0.07 3.65 3.11 4.33 47.65 47.07 47.56 1.41 1.15 0.80 92 94 106
Time after injection —Blood removed by bleeding —Blood retained in organs —Leg bones (tibias + femurs) —Liver —Gizzard —Intestine —Caeca —Yolk sac —Heart —Spleen —Feet (tarsals + phalanges) —Residual carcass —Feces Total
916
J . GODET AND M . BELHANI
—during the first day, radioactive iron accumulates in the organs mentioned above. Maximum activity was reached during the sixth hour for the liver, the
eighth for the marrow and the twelfth for the digestive tract; —during the second day, most of the iron which had accumulated, i.e. 29% of the injected radioactivity, left the marrow, the liver and the digestive tract whose radioactivity decreased respectively of 77%, 64% and 80%. During that time, 25% of the injected radioactivity was transferred into the blood stream; —after the second day, the radioactivity of the marrow, of the liver and of the digestive tract decreased only slightly. Blood radioactivity continued to increase but by only 7% per day. As far as the blood is concerned (Table 2), the radioactivity of plasma is always less than 1% of the injected radioactivity after the sixth hour. On the other hand, the radioactivity of hemoglobin increases rapidly, particularly during the first two days, to reach after 6 days 47% of the injected radioactivity. The non-hemoglobin components of the red blood cells take up 4 to 5% of the injected radioactivity, 10% of which is bound to components, present in the hemolysates and eluted first during gel filtration on Sephadex G-75. DISCUSSION There are two possible explanations for the retention of 59 Fe in the yolk sac after intravitelline injection. Either the embryonic
TABLE 2.—Blood distribution of radioactivity over a week period after intraperitoneal injection of
59
Fe
% of injected radioactivity Time after injection 6 hours 12 hours 24 hours 48 hours 6 days
Whole blood 1 10.22 11.86 15.91 39.92 50.99
Plasma 2 0.47 1.75 0.86 0.17 0.13
Red blood cells 3 9.75 10.11 15.05 39.75 50.86
Hemoglobin 4 4.17 5.59 10.52 36.64 47.41
Red blood cell non-hemoglobin components 5 5.58 4.52 4.52 3.11 3.45
1, 2, 3 : determined by direct measure of known aliquots 4 : calculated from the specific activity of purified hemoglobin and blood hemoglobin content 5 : calculated as the difference between red blood cell and purified hemoglobin radioactivity All values are corrected for total blood volume.
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
blood, leg bone marrow (tibias and femurs), liver and digestive tract (gizzard, intestine, coeca). In the above, the erythron alone (blood + bone marrow) contains 60% of the radioactivity, i.e. 24% of the total injected radioactivity. Each one of the other separated organs (heart, spleen, yolk sac) never contains more than 1% of the injected radioactivity, with the exception of the feet (tarsals and phalanges) whose radioactivity represents about 4% of the administered 59 Fe. About 1% of the injected radioactivity is eliminated with feces. Intravitelline injection slows down to a large extent the distribution of 59 Fe in the organs. Almost 90% of the radioiron administered in this way is still present in the yolk sac three to six hours later. Blood, bone marrow, liver and digestive tract contain, at that time, less than 4% of the injected radioactivity. After the fourth hour and during the first six days after injection, approximately 50% of the injected radioactivity stays in the blood, the bone marrow, the liver and the digestive tract (Figure 1). 59 Fe exchanges between these organs occured in three steps as follows:
'FE DISTRIBUTION
Since, in chicks, part of the erythropoietic tissues is found not only in the leg Femes but also in the other long bones and the back bone, it was observed that, soon after the intraperitoneal injection of 59 Fe, most of the radioactivity was divided between the marrow, the blood, the liver and the digestive tract. This distribution is observed in most mammals: various analyses have indeed shown that, for mice and rats, more than 90% of the injected iron is found after six hours in these four organs (Konitzer and Michalke, 1965; Cheney et al., 1967). Further, inhens(Rusov, 1965;Ramsay, 1966)and adult rodents (Konitzer and Michalke, 1965; Cheney et al., 1967) maximum radioactivity is observed after 6 hours in the marrow and 10 days in the blood stream. Hence, no major differences are found in radioiron exchanges between poultry and mammals. There are differences in the variations of the marrow and blood radioactivity between chicks injected at 3 days of age and adult hens. According to Rusov (1965), marrow radioactivity of the hen is lower than blood radioactivity in the first 5 days, while, in
chicks, the radioactivity in the blood is higher than in the marrow after the sixteenth hour. Similarly, Rusov (1965) shows that, for hens, 3.4% and 60% of the injected radioactivity is found in the blood respectively 24 hours and 6 days after injection, while the present work performed on chicks gives corresponding values of 15 and 50%. Thus, 59 Fe exchanges between marrow and blood occur from four to five times faster in 3 day old chicks than in adult hens. This difference can be accounted for by comparing the fast rate of blood formation in chicks which, from hemoglobin determination (Godet et al, 1970a), reaches 10% per day to the daily red blood cell renewal which, in adult hens, affects only 2% of the total blood. Table 2 shows that, in 3 day old chicks, 90% of the blood total radioactivity is found in hemoglobin 2 days after injection. Thus, 59 Fe exchanges between the various red blood cell compartments occur almost exclusively in order to synthesize hemoglobin. Approximately 4% of the injected radioactivity is bound very rapidly to the insoluble components of the stroma. This stroma-bound radioactivity shows no important variations during the week after injection and could result from an adsorption phenomenon similar to the one described, after in vitro experiments, by Clark (1967) in mature hen erythrocytes. The results presented above tend to show that the main steps in 59 Fe incorporation in 3 day old chicks are similar to those found in other laboratory animals. The differences observed between chicks and adult hens can undoubtedly be attributed to differences in blood formation rate inherent to age. It also appears that, in chicks as in most vertebrates, the newly administered iron is selectively used in preference to body iron already stored. The foregoing results and analysis led to an undertaking of a precise quantitative study of hemoglobin synthesis during chick development (Godet, 1973).
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
rudiment plays no part in iron metabolism after hatching and the iron is slowly absorbed along with other constituents or the yolk sac is still, 3 days after hatching, an important iron reserve which can be mobilized for erythropoiesis and in which the injected iron is diluted. Figure 1 shows that during the two days after intraperitoneal injection, most of the 59 Fe required for erythropoiesis is taken primarily from the liver and also, but for much smaller amounts, from the digestive tract, as the loss of radioactivity through excretion cannot alone justify the radioactivity loss observed in this organ. During the next four days, 10% of the injected radioactivity enters the blood stream which suggests that approximately 2 to 3% of the injected radioactivity is taken daily from organs other than the liver and the digestive tract.
917
918
J . GODET AND M . BELHANI
REFERENCES
Effects of Various Lighting Regimes on Diurnal Rhythms of EEG Components in the Chicken JIRO Y A N O , SHUNZO OSHIMA AND JIRO GOTOH
Department of Animal Physiology, Faculty of Agriculture, Nagoya University, Nagoya,
Japan
(Received for publication July 17, 1973)
ABSTRACT Effects of different photoperiods on the appearance of EEG slow waves were examined in freely-moving chickens by a radio telemetry system. The experiments were performed under 14L10D, 18L6D, 21L3D and 24L. It was clear that the EEG components were strictly synchronized to light-dark cycles. Continuous illumination exerted a dampening effect on the appearance of the slow wave diurnal rhythms. Chickens exposed to light-dark cycles of 14L10D and 18L6D maintained a constant daily level of slow wave activity. These levels are regarded as a normal amount of slow wave activity in male chickens. The daily amount of slow wave activity under 21L3D and 24L is probably regulated in a way different from that under 14L10D and 18L6D. The illumination seems to exert a strong effect on the mechanism controlling the appearance of the EEG in chickens when compared to mammals. POULTRY SCIENCE 53: 918-923, 1974
T
HERE are a number of publications concerning the electroencephalograph (EEG) during sleep and wakefulness in birds (Ookawa and Gotoh, 1964; Ookawa and Kadono, 1968; Ookawa, 1972). The existence of 3 stages of sleep-wakefulness, i.e. wakefulness, slow sleep (SS)and paradoxical sleep (PS), has been established in chickens and Japanese quail as well as mammals. The PS appears periodically between the SS but its proportion to total sleep time does not exceed a few percent in chickens (Gotoh, 1968). This may suggest that the EEG of the chicken can be tentatively classified into two major patterns, fast wave and slow wave.
It is well known that sleep-wakefulness shows diurnal rhythms synchronized to daynight cycles. Investigations of diurnal rhythm of EEG components have been undertaken with cats under continuous illumination (Sterman et al, 1965; Chase and Sterman, 1967) and with rats (Colvin et al, 1968) and rabbits (Spie et al, 1970) under light-dark cycles. These reports showed diurnal fluctuations of EEG components. In birds, most investigations on the biological rhythm have been carried out from such viewpoints as migration, homing instinct and photoperiodism. Cain and Wilson(1971,1972) reported that the locomotor activities of the
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
Cheney, B. A., K. Lothe, E. H. Morgan, S. K. Sood and C. A. Finch, 1967. Internal iron exchange in the rat. Am. J. Physiol. 212: 376-380. Clark, P., 1967. Uptake of iron by mature erythrocytes. Aust. J. Exp. Biol. Med. Sci. 45: 97-104. Godet, J., 1973. Synthese postnatale d'hemoglobine F chez le poulet. Comp. Rend. Acad. Sci. 276: 1201-1204. Godet, J., D. Schiirch, J. P. Blanchet and V. Nigon, 1970a. Evolution des caracteristiques erythrocytaires au cours du developpement post-embryonnaire du poulet. Exp. Cell Res. 60: 157-165.
Godet, J., D. Schiirch and V. Nigon, 1970b. Caracterisation et evolution des hemoglobines dans le cours du developpement postembryonnaire chez la poule. J. Embry. Exp. Morph. 23: 153-167. Konitzer, K., and K. Michalke, 1965. Der eisenstoffweschel der weissen Mous Ausscheichung und organ verterlung einen Fe59 tracer dosis. Acta Biol. Med. Germ. 14: 489-495. Ramsay, W. N. M., 1966. The incorporation of iron into hemoglobin in the domestic fowl. Quat. J. Exp. Physiol. 51: 221-228. Rusov, C , 1965. Recherches sur la ferrocinetique des volailles normales par Fe59. Med. Landbow. Opzechugs Genet. 30: 787-795.
Section de Biologie Generate et Appliquee Laboratoire Associe au C.N.R.S., Universite Claude Bernard de Lyon, 43, boulevard du 11 novembre 1918 69621 Villeurbanne, France (Received for publication July 14, 1973)
POULTRY SCIENCE 53: 914-918, 1974
JN VIVO labelling of hemoglobin by 59Fe -t-involves all of the complex mechanisms of storage and exchange of this radioactive isotope in and between the various parts of the organism. These mechanisms are relatively unknown in poultry which have not reached the adult stage. Hence, to study avian hemoglobin synthesis during post-hatching life, it became necessary to define the main pathways of radioiron uptake in the developing chick. 59Fe was thus injected, in different ways, to 3 day old chicks and the subsequent labelling of various organs was followed for a week. Attention was focused mainly on the interactions, between the internal exchange of 59Fe and its incorporation into hemoglobin, which occured during that week. MATERIAL AND METHODS White Leghorn young chicks weighing 35-45 g. received by intraperitoneal or intravitelline injection 50 to 100 ux. of 59Fe (ferric citrate; average specific activity 12 mCi./mg. Fe). The exact amount of radioactivity injected was determined by weighing both the aliquot of the material used as a standard and the amount injected into the
'Present address: Centre Pierre et Marie Curie, Service d'Hematologie, Alger.
animal. At the time of sacrifice, the body weight of each animal was determined and a measured volume of blood was withdrawn from the wing vein. A first fraction of this blood was used to measure, as described elsewhere (Godet et al., 1970 a), blood hemoglobin content and hematocrit. A second fraction was used to measure the radioactivity of known aliquots of whole blood, plasma and red blood cells, after, in the case of red blood cells, five washes with NaCl 0.9%. The rest of the collected blood was used to prepare hemolysates from which hemoglobin was purified by gel filtration on Sephadex G-75 (Godet et al, 1970b). Once killed by decapitation, each animal was carefully dissected. Easily separated organs, like tibia, femur, spleen, liver, etc . . . were placed directly in counting tubes, the large organs being first cut in small pieces. The remaining carcass which includes skeleton, muscles, skin and feathers was first coarsely ground and then distributed in counting tubes. Radioactivity of the various organs, of blood and feces was measured in a Packard Automatic Gamma Counter with less than 5% variation. The results were expressed as percent of injected radioactivity and corrections were made, as far as organs are concerned, for the activity due to their blood content. The organ blood content was calculated from organ weight and total blood volume. The
914
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
ABSTRACT In order to study avian hemoglobin synthesis, 59Fe distribution and exchanges between various organs and erythrocyte components have been examined after injection in 3 day old chicks. Results obtained during the week after injection show that the overall ferrokinetics follow the same trends as those observed in most animals. The only differences noted can be attributed to the rapid growth of the chicks at that age.
59
915
F E DISTRIBUTION
TABLE 1.—Organ radioactivity after intraperitoneal injection of S9Fe
total blood volume was, in turn, calculated from the body weight (Godet et al., 1970a). RESULTS Animals, sacrificed at different times during the 24 hours which follow injection, show
that the amount of 59Fe recovered represents within ± 8% the amount of administered radioactivity (Table 1). After intraperitoneal injection, about 40% of the administered 59 Fe is found, four hours later, in a group of tissues formed by the
% o t injected radioactivity
40
20.
4h
12h
24 h
»«»...»„...//.*.... —' // 48h
•
" 6d time after injection
FIG. 1. 59Fe exchanges between blood, bone marrow, liver and digestive tract, over a week period after injection to 3 day old chicks
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
% of injected radioactivity 8 hours 4 hours 24 hours 11.90 4.46 \ } 26.35 5.45 / 9.90 14.45 5.*>">» 13.08 11.40 15.12 6.34 8.11 5.77 4.161 4.60 4.34) 4.00 > 9.05 1.94 4.17 > 9.52 6.80 0.89; 0.26 1.01 ) 0.06 0.38 0.04 0.61 0.73 0.59 0.12 0.06 0.07 3.65 3.11 4.33 47.65 47.07 47.56 1.41 1.15 0.80 92 94 106
Time after injection —Blood removed by bleeding —Blood retained in organs —Leg bones (tibias + femurs) —Liver —Gizzard —Intestine —Caeca —Yolk sac —Heart —Spleen —Feet (tarsals + phalanges) —Residual carcass —Feces Total
916
J . GODET AND M . BELHANI
—during the first day, radioactive iron accumulates in the organs mentioned above. Maximum activity was reached during the sixth hour for the liver, the
eighth for the marrow and the twelfth for the digestive tract; —during the second day, most of the iron which had accumulated, i.e. 29% of the injected radioactivity, left the marrow, the liver and the digestive tract whose radioactivity decreased respectively of 77%, 64% and 80%. During that time, 25% of the injected radioactivity was transferred into the blood stream; —after the second day, the radioactivity of the marrow, of the liver and of the digestive tract decreased only slightly. Blood radioactivity continued to increase but by only 7% per day. As far as the blood is concerned (Table 2), the radioactivity of plasma is always less than 1% of the injected radioactivity after the sixth hour. On the other hand, the radioactivity of hemoglobin increases rapidly, particularly during the first two days, to reach after 6 days 47% of the injected radioactivity. The non-hemoglobin components of the red blood cells take up 4 to 5% of the injected radioactivity, 10% of which is bound to components, present in the hemolysates and eluted first during gel filtration on Sephadex G-75. DISCUSSION There are two possible explanations for the retention of 59 Fe in the yolk sac after intravitelline injection. Either the embryonic
TABLE 2.—Blood distribution of radioactivity over a week period after intraperitoneal injection of
59
Fe
% of injected radioactivity Time after injection 6 hours 12 hours 24 hours 48 hours 6 days
Whole blood 1 10.22 11.86 15.91 39.92 50.99
Plasma 2 0.47 1.75 0.86 0.17 0.13
Red blood cells 3 9.75 10.11 15.05 39.75 50.86
Hemoglobin 4 4.17 5.59 10.52 36.64 47.41
Red blood cell non-hemoglobin components 5 5.58 4.52 4.52 3.11 3.45
1, 2, 3 : determined by direct measure of known aliquots 4 : calculated from the specific activity of purified hemoglobin and blood hemoglobin content 5 : calculated as the difference between red blood cell and purified hemoglobin radioactivity All values are corrected for total blood volume.
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
blood, leg bone marrow (tibias and femurs), liver and digestive tract (gizzard, intestine, coeca). In the above, the erythron alone (blood + bone marrow) contains 60% of the radioactivity, i.e. 24% of the total injected radioactivity. Each one of the other separated organs (heart, spleen, yolk sac) never contains more than 1% of the injected radioactivity, with the exception of the feet (tarsals and phalanges) whose radioactivity represents about 4% of the administered 59 Fe. About 1% of the injected radioactivity is eliminated with feces. Intravitelline injection slows down to a large extent the distribution of 59 Fe in the organs. Almost 90% of the radioiron administered in this way is still present in the yolk sac three to six hours later. Blood, bone marrow, liver and digestive tract contain, at that time, less than 4% of the injected radioactivity. After the fourth hour and during the first six days after injection, approximately 50% of the injected radioactivity stays in the blood, the bone marrow, the liver and the digestive tract (Figure 1). 59 Fe exchanges between these organs occured in three steps as follows:
'FE DISTRIBUTION
Since, in chicks, part of the erythropoietic tissues is found not only in the leg Femes but also in the other long bones and the back bone, it was observed that, soon after the intraperitoneal injection of 59 Fe, most of the radioactivity was divided between the marrow, the blood, the liver and the digestive tract. This distribution is observed in most mammals: various analyses have indeed shown that, for mice and rats, more than 90% of the injected iron is found after six hours in these four organs (Konitzer and Michalke, 1965; Cheney et al., 1967). Further, inhens(Rusov, 1965;Ramsay, 1966)and adult rodents (Konitzer and Michalke, 1965; Cheney et al., 1967) maximum radioactivity is observed after 6 hours in the marrow and 10 days in the blood stream. Hence, no major differences are found in radioiron exchanges between poultry and mammals. There are differences in the variations of the marrow and blood radioactivity between chicks injected at 3 days of age and adult hens. According to Rusov (1965), marrow radioactivity of the hen is lower than blood radioactivity in the first 5 days, while, in
chicks, the radioactivity in the blood is higher than in the marrow after the sixteenth hour. Similarly, Rusov (1965) shows that, for hens, 3.4% and 60% of the injected radioactivity is found in the blood respectively 24 hours and 6 days after injection, while the present work performed on chicks gives corresponding values of 15 and 50%. Thus, 59 Fe exchanges between marrow and blood occur from four to five times faster in 3 day old chicks than in adult hens. This difference can be accounted for by comparing the fast rate of blood formation in chicks which, from hemoglobin determination (Godet et al, 1970a), reaches 10% per day to the daily red blood cell renewal which, in adult hens, affects only 2% of the total blood. Table 2 shows that, in 3 day old chicks, 90% of the blood total radioactivity is found in hemoglobin 2 days after injection. Thus, 59 Fe exchanges between the various red blood cell compartments occur almost exclusively in order to synthesize hemoglobin. Approximately 4% of the injected radioactivity is bound very rapidly to the insoluble components of the stroma. This stroma-bound radioactivity shows no important variations during the week after injection and could result from an adsorption phenomenon similar to the one described, after in vitro experiments, by Clark (1967) in mature hen erythrocytes. The results presented above tend to show that the main steps in 59 Fe incorporation in 3 day old chicks are similar to those found in other laboratory animals. The differences observed between chicks and adult hens can undoubtedly be attributed to differences in blood formation rate inherent to age. It also appears that, in chicks as in most vertebrates, the newly administered iron is selectively used in preference to body iron already stored. The foregoing results and analysis led to an undertaking of a precise quantitative study of hemoglobin synthesis during chick development (Godet, 1973).
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
rudiment plays no part in iron metabolism after hatching and the iron is slowly absorbed along with other constituents or the yolk sac is still, 3 days after hatching, an important iron reserve which can be mobilized for erythropoiesis and in which the injected iron is diluted. Figure 1 shows that during the two days after intraperitoneal injection, most of the 59 Fe required for erythropoiesis is taken primarily from the liver and also, but for much smaller amounts, from the digestive tract, as the loss of radioactivity through excretion cannot alone justify the radioactivity loss observed in this organ. During the next four days, 10% of the injected radioactivity enters the blood stream which suggests that approximately 2 to 3% of the injected radioactivity is taken daily from organs other than the liver and the digestive tract.
917
918
J . GODET AND M . BELHANI
REFERENCES
Effects of Various Lighting Regimes on Diurnal Rhythms of EEG Components in the Chicken JIRO Y A N O , SHUNZO OSHIMA AND JIRO GOTOH
Department of Animal Physiology, Faculty of Agriculture, Nagoya University, Nagoya,
Japan
(Received for publication July 17, 1973)
ABSTRACT Effects of different photoperiods on the appearance of EEG slow waves were examined in freely-moving chickens by a radio telemetry system. The experiments were performed under 14L10D, 18L6D, 21L3D and 24L. It was clear that the EEG components were strictly synchronized to light-dark cycles. Continuous illumination exerted a dampening effect on the appearance of the slow wave diurnal rhythms. Chickens exposed to light-dark cycles of 14L10D and 18L6D maintained a constant daily level of slow wave activity. These levels are regarded as a normal amount of slow wave activity in male chickens. The daily amount of slow wave activity under 21L3D and 24L is probably regulated in a way different from that under 14L10D and 18L6D. The illumination seems to exert a strong effect on the mechanism controlling the appearance of the EEG in chickens when compared to mammals. POULTRY SCIENCE 53: 918-923, 1974
T
HERE are a number of publications concerning the electroencephalograph (EEG) during sleep and wakefulness in birds (Ookawa and Gotoh, 1964; Ookawa and Kadono, 1968; Ookawa, 1972). The existence of 3 stages of sleep-wakefulness, i.e. wakefulness, slow sleep (SS)and paradoxical sleep (PS), has been established in chickens and Japanese quail as well as mammals. The PS appears periodically between the SS but its proportion to total sleep time does not exceed a few percent in chickens (Gotoh, 1968). This may suggest that the EEG of the chicken can be tentatively classified into two major patterns, fast wave and slow wave.
It is well known that sleep-wakefulness shows diurnal rhythms synchronized to daynight cycles. Investigations of diurnal rhythm of EEG components have been undertaken with cats under continuous illumination (Sterman et al, 1965; Chase and Sterman, 1967) and with rats (Colvin et al, 1968) and rabbits (Spie et al, 1970) under light-dark cycles. These reports showed diurnal fluctuations of EEG components. In birds, most investigations on the biological rhythm have been carried out from such viewpoints as migration, homing instinct and photoperiodism. Cain and Wilson(1971,1972) reported that the locomotor activities of the
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on June 7, 2015
Cheney, B. A., K. Lothe, E. H. Morgan, S. K. Sood and C. A. Finch, 1967. Internal iron exchange in the rat. Am. J. Physiol. 212: 376-380. Clark, P., 1967. Uptake of iron by mature erythrocytes. Aust. J. Exp. Biol. Med. Sci. 45: 97-104. Godet, J., 1973. Synthese postnatale d'hemoglobine F chez le poulet. Comp. Rend. Acad. Sci. 276: 1201-1204. Godet, J., D. Schiirch, J. P. Blanchet and V. Nigon, 1970a. Evolution des caracteristiques erythrocytaires au cours du developpement post-embryonnaire du poulet. Exp. Cell Res. 60: 157-165.
Godet, J., D. Schiirch and V. Nigon, 1970b. Caracterisation et evolution des hemoglobines dans le cours du developpement postembryonnaire chez la poule. J. Embry. Exp. Morph. 23: 153-167. Konitzer, K., and K. Michalke, 1965. Der eisenstoffweschel der weissen Mous Ausscheichung und organ verterlung einen Fe59 tracer dosis. Acta Biol. Med. Germ. 14: 489-495. Ramsay, W. N. M., 1966. The incorporation of iron into hemoglobin in the domestic fowl. Quat. J. Exp. Physiol. 51: 221-228. Rusov, C , 1965. Recherches sur la ferrocinetique des volailles normales par Fe59. Med. Landbow. Opzechugs Genet. 30: 787-795.