ONTOGENY OF CHICKEN HEMOGLOBIN. III. IMMUNOLOGICAL STUDY OF THE HETEROGENEITY OF HEMOGLOBIN IN DEVELOPMENT

By applying the double diffusion technique of Ouchterlony and the immunoelectrophoresis, sequence in the appearance of antigens reactive with antisera against HbCOs from 5-day embryos and adult chickens, and the major component of adult HbCO in the course of chicken development has been studied.
Antigenic components reacting to antiserum against HbCO from 5-day embryos have been detected throughout development. Three globin-like components specific to early embryos are detectable in embryos to 2 days of incubation. Two Hb components are detectable in embryos from 3 to 5 days of incubation; one is apparently specific to embryos, while the other seems to be somewhat different from adult Hb. After 6 days of incubation only one adult Hb component is detectable.
Antigenic components reacting to antiserum against HbCO from adults have also been detectable throughout embryonic life. One globin-like component specific to early embryos can be found in embryos to 2 days of incubation. One or two Hb components which are probably specific to embryos can be detected in embryos from 3 to 5 days of incubation. After 6 days of incubation one adult Hb component is detectable, while one globin-like component specific to adults can be found after 15 days of incubation. Further, the other globin-like component is detectable after 3 days of incubation.
Antigenic components reacting to antiserum against the major component of adult HbCO have been detected throughout development; one which seems to be somewhat different from either adult Hb components can be found in embryos from 2 to 5 days of incubation, while the other which is identical with the major component of adult Hb is detectable after 6 days of incubation.     

Effects of lifestyle and genetic polymorphisms on consumption of coffee or black tea and urinary caffeine levels

To find the cause of individual differences in caffeine intake and its metabolism, we investigated the effects of lifestyle and genetic polymorphisms of caffeine metabolic enzymes on coffee or black tea and urinary caffeine levels among 259 male Japanese. It was seen that cigarette smokers drank more coffee or black tea than non-smokers (p < 0.001). There was an inverse correlation between the amount of coffee or black tea consumed and age or the frequency of alcohol drinking (p < 0.05). Genetic polymorphisms of N -acetyltransferase 2 (NAT2), cytochrome P450 (CYP)1A1 and 2E1 did not significantly affect the habit of drinking coffee or black tea. The frequency of allele 1, the NAT2 allele of rapid acetylators, increased according to coffee or black tea consumption (0.05 < p < 0.1). Among lifestyle factors, two factors, i.e. smoking and the amount of coffee or black tea consumed, were related to urinary caffeine levels (p < 0.05). Geometric means of urinary caffeine levels were higher in the group who consumed higher amounts of coffee or black tea (p < 0.05) and those of smokers were lower than non-smokers- approximately 70% of non-smokers (p < 0.05). The genetic polymorphisms of NAT2, CYP1A1 and CYP2E1 were not significantly associated with the urinary caffeine levels according to each consumption level of coffee or black tea. This study suggests that smoking should be considered for the proper appreciation of individual differences in caffeine intake and urinary caffeine levels.     

Morph‐dependent form of asymmetry in mandibles of the stag beetle Prosopocoilus inclinatus (Coleoptera: Lucanidae)

Abstract 1. The form of asymmetry in bilateral organs usually follows the same pattern within single populations. However, some exceptions may occur when a population consists of different phenotypes that are from different ontogenic backgrounds and under different selective pressures. We investigated the asymmetric patterns of mandibles of larvae, females, and males in the stag beetle Prosopocoilus inclinatus. 2. Larval mandibles exhibited directional asymmetry both in length and cross direction, whereas female mandibles showed directional asymmetry in cross direction. These asymmetric structures might be more effective in cutting wood fibres. 3. For the relation of male mandible length to body size, a model with a switch point showed a better fit to the data than a convex curve model. This shows that the males are dimorphic with two distinct morphs. 4. The form of asymmetry in male mandible length differed between the morphs. The smaller males exhibited left‐biased directional asymmetry in common with larvae, whereas the larger males exhibited fluctuating asymmetry. 5. This is a novel finding of a morph‐dependent asymmetry. The morph‐dependent asymmetry in males may be as a result of different selection on each morph or a developmental constraint from larval mandibles to adult ones.     

Morphological Characterization of Three Phenotypes of the Isopod Armadillidium vulgare

The morphological characteristics and ommochrome quantity in the integument of red, white, and wild type (black-grey) Armadillidium vulgare were studied. The red phenotype was found to possess two kinds of immature ommochrome pigment granules within its pigment cells, in addition to mature pigment granules. The immature granules seemed to contain uniformly distributed fibrilles, or to have an electron-dense central region surrounded by an electron-lucent outer edge. Since these immature pigment granules were typically observed to be distributed along with the mature ones, and were also more easily extractable than the wild type's, it is hypothesized that ommochrome granule maturation in the red phenotype may occur slowly due to a defect in the pigment granule internal process which combines pigments with matrix proteins. Regarding the white phenotype, although its pigment cells were undeveloped, several large-sized vesicles containing a small amount of electron-dense material appeared in the pigment cell cytoplasm. The wild and red type males of A. vulgare were found to have an ommochrome content twice as large as that of the corresponding females, with no ommochrome pigment being detected in the white phenotype. The genetic relationship between the white and red phenotypes was discussed using as a basis the observed pigment granule structure.     

Involvement of Pteridines in the Body Coloration of the Isopod Armadillidium vulgare

The pteridine content was measured as a function of age in Armadillidium vulgare, and the fine structure of the pteridine-containing granules in the integument was examined in relation to pteridine content. Yellow chromatophores are an essential component of the cream-markings, which are a defining feature of the female A. vulgare. Four kinds of pteridines in the integument including a yellow pigment (sepiapterin) were determined by HPLC. The body color of the red phenotype of A. vulgare varies from dark red to yellowish red and was formerly thought to be due to the quality and quantity of ommochrome pigment. Our analysis of the pteridine content in the integument of this phenotype revealed a significant change in sepiapterin content per body weight with age. Sepiapterin content per body weight decreased gradually with age, while that of biopterin tended to increase with age. Ultrastructural observations of the pigment granules in the yellow chromatophores revealed a corresponding change in the fine structure of pigment granules. In the older adults, some of the electron-dense fibrous materials in the pteridine-containing granules was concentrically arranged, and in the younger adults, most of pteridine-containing granules were electron-lucent. The role of pteridine quality in determining the structure of pteridine-containing granules is discussed.     

Coexistence of β1- and β2-Adrenoceptors in the Melanophore of the Goby Tridentiger obscurus*

The effects of β-adrenergic agonists and antagonists on the pigmentary state of denervated melanophores in isolated, split, caudal fins of the goby Tridentiger obscurus were examined to investigate the function and the subtype of the β-adrenoceptors of the melanophores. Salbutamol, terbutaline, and dobutamine partially inhibited the pigment-aggregating response of melanophores to norepinephrine. The effects of these β-agonists were inhibited by propranolol. It was confirmed that the melanophores possess both α-and β-adrenoceptors, and that the activation of the β-adrenoceptors induces the dispersion of pigment in the melanophores. Norepinephrine, epinephrine, isoproterenol, dobutamine, salbutamol, and terbutaline evoked the dispersion of pigment in the melanophores in which pigment had previously been aggregated by treatment with verapamil in the presence of phentolamine. The pigment-dispersing effects of two β₁-selective agonists, norepinephrine and dobutamine, were effectively inhibited by metoprolol, a selective antagonist of β₁-receptors. By contrast, the pigment-dispersing effects of two β₂-selective agonists, salbutamol and terbutaline, were not inhibited by metoprolol. Both the effects of nonselective agonists, epinephrine and isoproterenol, were partially inhibited by metoprolol. The actions of all of the β-agonists used were effectively inhibited by propranolol, and they were partially inhibited by butoxamine. These results suggest coexistence of β₁- and β₂-adrenoceptors in the melanophores. The relative numbers of β₁- and β₂-adrenoreceptors as a percentage of the total population of β-adrenoceptors were estimated to be 18.6% and 81.4%, respectively, from analyses of Hofstee plots of the effects of the β-agonists on the melanophores in the presence of butoxamine or metoprolol.     

Action Spectra for Step-Up Photophobic Response in Blepharisma

ABSTRACT The cells of Blepharisma which possess red pigment (blepharismin) show step-up photophobic response (temporal ciliary reversal induced by a sudden increase in light intensity). Bleaching of the cells by cold shock raised a threshold light intensity for the response, Oxidation of red pigment that produced blue pigment did not raise the threshold for the response. The action spectrum for the step-up photophobic response of the cells which possess normal red pigment had peaks at about 580, 540 and 490 nm, a value which coincided with peaks of an absorption spectrum of the red pigment. The absorption spectrum of oxidized pigment (blue pigment) shifted 20 nm toward infrared light. The action spectrum for the response of the cells which possess blue pigment also shifted 20 nm toward infrared light. Results suggest that red pigment might be involved in the step-up photophobic response. Key words. Blepharismin, ciliary reversal, photoreceptors, photoresponse.     

Variation of Abscisic Acid and Batatasin Content in Yam Bulbils--Effects of Stratification and Light Exposure

Batatasin content of yam (Dioscorea batatas Decne.) bulbilsdecreased slowly during 5 month stratification at 4°C, whileit remained virtually unchanged when stored at 23°C. Abscisicacid increased considerably during the 1st month at 23°Cbut only slightly at 4°C and thereafter remained constantup to the end of the 5-month storage period. On transfer to sprouting beds at 23°C the batatasin contentof stratified bulbils decreased sharply but that of non-stratifiedbulbils showed only slight variation. Light exposure duringsprouting tests suppressed the sprouting of the non-stratifiedbulbils but promoted that of the stratified ones. The changesin batatasin contents were found when the bulbils were exposedto light and this is discussed in relation to sprouting responsesto light. A scheme for batatasin economy in bulbil dormancyis proposed.     

ONTOGENY OF CHICKEN HEMOGLOBIN. I. ELECTROPHORETIC STUDY OF THE HETEROGENEITY OF HEMOGLOBIN IN DEVELOPMENT

Separation of different molecular species of hemoglobin from developing chickens by starch gel electrophoresis has revealed the appearance of early embryonic (embryonic), late embryonic (fetal) and adult hemoglobin (Hb) type during development. In 5-day embryos, there are 3 or 4 forms of embryonic Hb type. They begin to decrease in 6-day embryos and cannot be detected in embryos after 10 days of incubation. In 6-day embryos, two forms of adult Hb type appear, and one of them, which is a major form in adults, becomes t o be a major one in 7-day embryos. One or two forms of fetal Hb type first appear in 10-day embryos and are still present in 5-day posthatching chickens.
Ultracentrifugation of carbonmonoxyhemoglobins from embryos at early and at later stages (fetuses), from newly hatched and from adult chickens has shown that they have a single monodisperse peak. Some heterogeneity, however, has been detected after starch gel electrophoresis, probably owing to aggregation or polymerization.
Subunit analysis of embryonic, fetal and adult Hb type by starch gel electrophoresis in formate buffer at pH 1.9 has indicated that embryonic Hb type contains total 5 subunits, C, D, E, F and G; fetal Hb type, total 2, A and H; and adult Hb type, total 3, B, F and H.