Serum corticosterone concentrations in developing shell-less and shelled turkey embryos.

1. Turkey embryos grown in shell-less culture (EO) display normal development to day 14 of incubation, after which growth rate is reduced and mortality increases, compared to age-matched in ovo (IO) embryos (Richards 1982. Long term shell-less culture of turkey embryos. Poultry Sci. 61, 2089-2096). 2. In this study serum corticosterone concentrations were monitored in normally incubated embryos and embryos maintained in shell-less culture. 3. On days 14 and 16 of incubation, corticosterone levels were greater (P less than 0.05) in EO than IO embryos, whereas during the later stages corticosterone increased dramatically in the shelled embryos, and remained relatively constant in the EO embryos. 4. EO embryos do not exhibit a pattern of adrenal hormonal secretion that would indicated a stressful condition. 5. The absence of a normal increase in corticosterone in shell-less embryos may contribute to the abnormal embryological development in the later stages of incubation.     

Fine structure of chick embryonic parathyroid glands cultured on media with different concentrations of calcium.

An electron microscopical study of parathyroid glands from 13-day old chick embryos cultured for 2 days on media with different concentrations of calcium was conducted. The same as in non-cultured embryonic glands, the cells in all cultures contained a moderate amount of rough endoplasmic reticulum and a large Golgi complex with many cisternae, vacuoles, vesicles, coated vesicles and small prosecretory granules. In addition, large secretory granules, which are very rare in the non-cultured controls appeared frequently in the cultures and were especially numerous in the glands cultured on high-calcium medium. The fact that the amount of secretory granules varied according to the concentration of calcium in the medium is interpreted as indicating that the rate of parathyroid hormone secretion in the embryo is already responsive to variations in the concentration of calcium. To the extent that in vitro results may be accepted as representative of what happens in vivo these results support the idea that the embryonic gland may be controlled by variations in the calcemia as the adult gland does.     

A diabetic-like condition of turkey embryos maintained in shell-less culture

Serum insulin concentration and pancreatic insulin content were determined for turkey embryos incubated in ovo and in long-term shell-less culture (ex ovo). Insulin was undetectable (less than 10 pg) in serum from 87% of the ex ovo embryos compared with their in ovo counterparts. This was evident at all incubation ages, although insulin was detectable in more of the ex ovo embryos on Day 24. Insulin increased in the embryos incubated in ovo from 122 (Day 15) to levels exceeding 2000 pg/ml at hatching. Total pancreatic insulin content was greater in the cultured embryos on Days 15, 17, and 22 compared with their in ovo counterparts. Serum glucose was significantly greater (P less than 0.05) in the ex ovo embryos at all ages. In response to an infusion of L-arginine, serum insulin increased from 566 to 1256 pg/ml in the in ovo embryos, whereas no change was evident in the ex ovo embryos (233 vs 257 pg/ml). When embryos incubated in ovo were injected with insulin, a significant (P less than 0.05) reduction of serum glucose was observed at 60 min after injection. Serum glucose concentrations remained elevated in the embryos incubated ex ovo despite the insulin injection. Liver glucose 6-phosphatase activity, assessed on Days 15 and 22 of incubation, was found to be significantly (P less than 0.05) lower in the ex ovo embryos. Turkey embryos incubated in shell-less culture exhibited chronic hyperglycemia in concert with extremely low circulating levels of insulin. The pancreatic beta cells of these embryos were not responsive to arginine or elevated glucose. Taken together these findings suggest the occurrence of a diabetic-like condition in the ex ovo embryos. This defect in insulin secretion may, in part, be responsible for some of the developmental abnormalities characteristic of the turkey embryo cultured ex ovo.     

Effect of a low calcium diet on the ultrastructure of the parathyroid gland of chicks

Summary The effect of a low calcium diet on the ultrastructure of the parathyroid gland in the chick was examined. Two-week-old White Leghorn chicks were fed a low calcium diet (calcium content 0.63%) for two weeks. In these chicks, the parathyroid glands are grossly enlarged. Hypertrophy and hyperplasia of the chief cells are evident. The plasma membranes between adjacent cells are relatively straight but interdigitate in some places. Chief cells contain occasional membrane-limited secretory granules (150–350 m in diameter) and with contents of variable electron density. Secretory granules are distributed randomly but some are closely applied to the plasma membrane. There is an increase in the development of the rough-surfaced endoplasmic reticulum. The Golgi complex is enlarged and consists of cisternae arranged in concentric layers, smooth-surfaced and coated vesicles and condensing vacuoles. Dilatations of the cisternae at several points are observed. Mitochondria and filaments are also encountered. These morphological features suggest that low calcium intake stimulates the synthetic activity of the chief cells of the chick parathyroid.     

Mineral metabolism in the developing turkey embryo--I. The effects of developmental age and shell-less culture on trace element contents of selected tissues.

1. Turkey embryos were incubated in ovo or in long-term shell-less culture (ex ovo) for 14, 18, 22 or 26 days. The embryos incubated ex ovo exhibited a progressive decline in the rate of growth and were hypocalcemic and hypoproteinemic compared to their in ovo counterparts from day 18 to day 26 of incubation. 2. The ratio of the concentrations of alpha-fetoprotein and albumin (AFP/A) in serum was determined for both groups of embryos. The AFP/A ratio may be useful as a biochemical index to stage avian embryonic development. Using this index it was concluded that ex ovo embryos exhibited a progressive developmental retardation compared to in ovo embryos. 3. Significant differences were observed in serum trace element concentrations for embryos incubated in ovo vs ex ovo. Most notably, serum copper concentration was significantly lower in ex ovo embryos on days 18 and 22 of incubation and significantly higher on day 26 of incubation compared to serum from embryos incubated in ovo. 4. Livers from embryos incubated ex ovo exhibited significant differences trace element levels compared to those incubated in ovo. By day 26 of incubation the concentration and total amount of zinc and iron were markedly elevated, whereas copper was greatly reduced in the livers of embryos incubated ex ovo compared to the corresponding in ovo levels. 5. Hearts from embryos incubated ex ovo contained less zinc and copper and more iron by day 26 of incubation than those from embryos incubated in ovo.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of melatonin on the ultrastructure of the golden hamster parathyroid gland.

Ultrastructural changes of the parathyroid glands of melatonin-treated golden hamsters were studied. Many chief cells in the parathyroid glands after 1 hour of administration of melatonin contained poorly-developed Golgi complexes associated with a few prosecretory granules and numerous lipid droplets as compared with those of the control animals. The morphology of the parathyroid glands after 5 hours of administration resembled that of the control animals. Many chief cells in the parathyroid glands after 24 hours of administration had well-developed Golgi complexes and cisternae of the granular endoplasmic reticulum, numerous prosecretory granules, a few lipid droplets and many secretory granules in the peripheral cytoplasm as compared with those of the control animals. The ultrastructure of the parathyroid glands after 48 hours of administration was almost similar to that of the control animals. It is considered that melatonin affects the secretory activity of the parathyroid gland.     

Ultrastructural study on the effects of hypophysectomy on the golden hamster parathyroid gland.

The ultrastructure of the parathyroid glands of hypophysectomized golden hamsters was studied. In the parathyroid glands of hypophysectomized animals the Golgi complexes and secretory granules were significantly decreased and large vacuolar bodies were significantly increased compared with those of the control animals. In addition, the chief cells contained a few prosecretory granules in the Golgi areas and a few secretory granules were present in the peripheral cytoplasm. These results suggest that the synthesis and release of parathyroid hormone may be suppressed in the parathyroid glands of the hypophysectomized animals.     

Effects of isoproterenol on the fine structure of the hamster parathyroid gland

Ultrastructural changes of the parathyroid glands of isoproterenol-treated golden hamsters were investigated. Many chief cells in the parathyroid glands after 5 and 10 minutes of administration of isoproterenol contain well-developed Golgi complexes and granular endoplasmic reticulum, numerous prosecretory granules, and many secretory granules in the peripheral cytoplasm as compared with the control animals. Many chief cells in the parathyroid glands after 1, 3, 6 and 12 hours of administration have poorly-developed Golgi complexes, granular endoplasmic reticulum, many secretory granules and numerous lipid droplets as compared with the control animals. The morphology of the parathyroid gland after 30 minutes and 24 hours of administration resembles that of the control animals. It is considered that isoproterenol affects the secretory activity of the parathyroid gland.     

Vitamin D and chick embryonic yolk calcium mobilization: identification and regulation of expression of vitamin D-dependent Ca2(+)-binding protein, calbindin-D28K, in the yolk sac

The developing chick embryo acquires calcium from two sources. Until about Day 10 of incubation, the yolk is the only source; thereafter, calcium is also mobilized from the eggshell. We have previously shown that during normal chick embryonic development, vitamin D is involved in regulating yolk calcium mobilization, whereas vitamin K is required for eggshell calcium translocation by the chorioallantoic membrane. We have studied here the biochemical action of 1,25-dihydroxy vitamin D3 in the yolk sac by examining the expression and regulation of the cytosolic vitamin D-dependent calcium-binding protein, calbindin-D28K. Two types of embryos are used for this study, normal embryos developing in ovo and embryos maintained in long-term shell-less culture ex ovo, the latter being dependent solely on the yolk as their calcium source. Our findings are (1) calbindin-D28K is expressed in the embryonic yolk sac, detectable at incubation Days 9 and 14; (2) the embryonic yolk sac calbindin-D28K resembles that of the adult duodenum in both molecular weight (Mr 28,000) and isoelectric point, as well as the presence of E-F hand Ca2(+)-binding structural domains; (3) systemic calcium deficiency caused by shell-less culture of chick embryos results in enhanced expression of calbindin-D28K in the yolk sac during late development; (4) yolk sac calbindin-D28K expression is inducible by 1,25-dihydroxy vitamin D3 treatment in vivo and in vitro; and (5) immunohistochemistry revealed that yolk sac calbindin-D28K is localized exclusively to the cytoplasm of the yolk sac endoderm. These findings indicate that the chick embryonic yolk sac is a genuine target tissue of 1,25-dihydroxy vitamin D3.     

Ultrastructural observations on the mechanism of secretion in the rat parathyroid after fluoride ingestion

Summary The secretory mechanism of the parathyroid glands of fluoride treated rats is evaluated ultrastructurally and compared to that of control rats. The principal difference between the two groups of rats concerns the rate of activity of the chief cells of the gland. In the control animals, these cells are predominantly inactive. In the fluoride-treated rats, they exhibit a more active stage of the secretory cycle. The active chief cells in rats treated with fluoride contain increased numbers of secretory granules. These granules are released into the perivascular spaces within cytoplasmic projections suggesting an apocrine-like mechanism for the secretion of parathyroid hormone. Secretory granules are observed free in the perivascular spaces and within the cytoplasm of capillary endothelial cells in the parathyroid glands.     

Mineral metabolism in the developing turkey embryo--II. The role of the yolk sac.

1. Turkey embryos were incubated in ovo or in long-term shell-less culture (ex ovo) for 14, 18, 22 or 26 days, at which time the concentrations of zinc, copper, iron, manganese and calcium in yolk and yolk sac membrane were determined. 2. Yolk manganese and calcium concentrations increased during incubation in ovo while the concentrations of zinc, copper and iron declined. The concentrations of zinc, copper and iron in yolk from ex ovo embryos did not decline. Yolk calcium concentration increased during incubation ex ovo, although to a much lesser extent than that observed in ovo. 3. The concentration of zinc, copper and iron declined in yolk sac tissue during incubation in ovo whereas no decline was observed for yolk sac tissue from ex ovo embryos. Yolk sac calcium and manganese concentrations increased during incubation in ovo and ex ovo, although the increase in calcium concentration for ex ovo yolk sac was much smaller than that observed in ovo. 4. A peak corresponding to metallothionein (MT) which bound both zinc and copper was isolated from yolk sac cytosol on day 14 of incubation in ovo using gel-permeation column chromatography. 5. Further fractionation of the MT peak by anion exchange chromatography revealed three metal-binding peaks designated MT-1, MT-2a and MT-2b. The majority of the zinc was bound to MT-2a and MT-2b whereas most of the copper was bound to a single peak (MT-2b). 6. The concentrations of zinc and copper in yolk sac cytosol reached a maximum on day 14 of incubation in ovo and declined through to day 28 (hatching).(ABSTRACT TRUNCATED AT 250 WORDS)     

Ultrastructural studies on the effects of hypergravity environment on the parathyroid glands in golden hamsters of different ages

The ultrastructure of the parathyroid glands of infantile, young, adult and senile golden hamsters subjected to a hypergravity environment was studied. In the parathyroid glands of 5-, 10- and 20-day-old, and 1- and 3-month-old golden hamsters exposed to a hypergravity environment, the Golgi complexes were significantly increased, and in 5-, 10- and 20-day-old, and 1- and 8-month-old animals exposed to a hypergravity environment, the cisternae of the granular endoplasmic reticulum appeared to be increased as compared to those of each control group. In addition, in centrifuged animals numerous prosecretory granules were observed in the Golgi areas, and many secretory granules were located in the peripheral cytoplasm. The ultrastructure of the parathyroid glands of 14-month-old centrifuged animals resembled that of 14-month-old control animals. These results suggest that the secretory activity of the parathyroid gland may be stimulated in infantile, young and adult golden hamsters subjected to a hypergravity environment and may not be stimulated in senile animals subjected to a hypergravity environment.     

Electron microscopic study of the parathyroid gland of the calcium-treated hamster subjected to hypergravity environment

The ultrastructure of the parathyroid glands of calcium-treated golden hamsters subjected to 5 gravity environment was studied. In the calcium-treated animals exposed to hypergravity environment, the Golgi complexes and cisternae of the granular endoplasmic reticulum were significantly decreased compared with those of the animals exposed to hypergravity environment only and appeared to increase compared with those of the calcium-treated animals, but were almost similar to those of the control animals. In addition, many chief cells contained some prosecretory granules in the Golgi areas, some secretory granules situated close to the plasma membrane and many lysosomes. The morphology of the parathyroid glands in the calcium-treated animals exposed to hypergravity environment resembled that of the control animals. These results suggest that the parathyroid glands suppressed by treatment of calcium and stimulated in response to hypergravity environment may indicate the secretory activity of the parathyroid glands of the control animals.     

Effects of pinealectomy on the ultrastructure of the golden hamster parathyroid gland

Ultrastructural changes of the parathyroid glands of pinealectomized golden hamsters were investigated. The main changes in the parathyroid glands 1 hour and 1 day after pinealectomy compared with the control and sham-operated groups were an increase of the Golgi complexes, cisternae of the granular endoplasmic reticulum and large vacuolar bodies. In addition, many chief cells contained numerous prosecretory granules in the Golgi areas and many secretory granules in the peripheral cytoplasm. The morphology of the parathyroid glands 7 and 30 days after pinealectomy resembled that of the control parathyroid glands. These results suggest that pinealectomy affects the secretory activity of the parathyroid gland.     

Ultrastructure of the parathyroid gland of the japanese lizard in the spring and summer season

The ultrastructure of the parathyroid glands of adult Japanese lizards (Takydromus tachydromoides) in the spring and summer season was examined. The parenchyma of the gland consists of chief cells arranged in cords or solid masses. Many chief cells contain numerous free ribosomes and mitochondria, well-developed Golgi complexes, a few lysosome-like bodies, some multivesicular bodies and relatively numerous lipid droplets. The endoplasmic reticulum is mainly smooth-surfaced. Cisternae of the rough endoplasmic reticulum are distributed randomly in the cytoplasm. Small coated vesicles of 700-800 Å in diameter are found occasionally in the cytoplasm, especially in the Golgi region. The chief cells contain occasional secretory granules of 150-300 nm in diameter that are distributed randomly in the cytoplasm and lie close to the plasma membrane. Electron dense material similar to the contents of the secretory granules is observed in the enlarged intercellular space. These findings suggest that the secretory granules may be discharged into the intercellular space by an eruptocrine type of secretion. Coated vesicles (invaginations) connected to the plasma membrane and smooth vesicles arranged in a row near the plasma membrane are observed. It is suggested that such coated vesicles may take up extracellular proteins. The accumulation of microfilaments is sometimes recognized. Morphological evidence of synthetic and secretory activities in the chief cells suggests active parathyroid function in the Japanese lizard during the spring and summer season.     

Hypothalamo-hypophyseal correlates in the chick embryo: thyrotropic activity of adenohypophyseal grafts from donor embryos of various ages. Effect or pretreatment with TRH

Adenohypophyses from 10 to 18-day-old chick embryonic donors grafted in 3 days chick embryos, thus disconnected from the hypothalamus, had a partially autonomous thyrotrophic activity. However this functional autonomy was greater in grafts from donors aged 10 or 11 days than from older embryos or from 11-day donors pretreated with T.R.H. before grafting. This strongly suggests that hypothalamo-adenohypophysis-thyroidal relationship establish normally between 11 and 12 days of embryonic life.     

Influence of Hydrocortisone on Chick Embryo Retina Development

Treatment of chick embryos in ovo with hydrocortisone-21-phosphate (a single dose of 150 micrograms) caused a marked reduction of retinal thymidine kinase activity 24 h later. The inhibitory effect was highest (65-70%) in 8-10-day-old embryos and declined with age, disappearing after day 15. It was accompanied by a reduction in thickness of the retinal layers. Adrenocorticotropic hormone (ACTH) treatment (10 micrograms daily for 2 days) also produced an age-dependent inhibitory effect on retinal thymidine kinase, whereas treatment with a single dose of 200 micrograms of metopirone, a compound that prevents the 11 beta-hydroxylation of steroid molecules in the adrenal glands, impeded the decrease in thymidine kinase activity that normally occurs in chick embryo retina after day 9 of development. In addition, metopirone prevented the inhibition exerted by ACTH on thymidine kinase activity but had no effect on the action of hydrocortisone.     

Electron-microscopic study of the parathyroid gland of epinephrine-treated golden hamsters subjected to hypergravity environment.

The ultrastructure of the parathyroid glands of golden hamsters subjected to 5-gravity environment after administration of epinephrine was studied. In the epinephrine-treated animals exposed to a hypergravity environment, the Golgi complexes associated with numerous prosecretory granules were significantly increased compared with those of the control, centrifuged and epinephrine-treated animals, as well as the cisternae of the granular endoplasmic reticulum compared with those of the control and centrifuged animals. In addition, many secretory granules were situated close to the the plasma membrane of the chief cells in the epinephrine-treated animals exposed to a hypergravity environment. Those observations suggest that the secretory activity of the parathyroid gland may be markedly stimulated in the epinephrine-treated animals exposed to a hypergravity environment.     

Ultrastructure of the parathyroid gland of fetal and pregnant golden hamsters subjected to hypergravity environment.

The ultrastructure of the parathyroid glands of fetal and pregnant golden hamsters exposed to 5-gravity environment was studied. In the centrifuged fetal animals the Golgi complexes associated with some prosecretory granules were significantly increased compared with those of the control fetal animals, and several secretory granules were located in a peripheral position just beneath the plasma membrane. In the centrifuged pregnant animals the Golgi complexes associated with numerous prosecretory granules were significantly increased, cisternae of the granular endoplasmic reticulum appeared to be increased compared with those of the control pregnant animals, many secretory granules were located in the peripheral cytoplasm and several granules were situated close to the plasma membrane. These findings suggest that in the parathyroid glands of the fetal golden hamsters as well as the pregnant animals the synthesis and release of parathyroid hormone may be stimulated in response to a hypergravity environment.     

Development of chicken epidermis cultured with embryo extract

The epidermis from 11-day-old chick embryo shank skin was cultured with 11-day-old chick embryo extract. The growth and the differentiation of the epidermis in culture were studied histologically, electron microscopically and with polyacrylamide gel electrophoresis of keratin proteins. The epidermis cultured with the chick embryo extract proliferated and stratum structures developed simultaneously with the increase in epidermal cell layers. Finally, a keratinized layer was observed after 10 days in culture. Electron microscopic observations revealed that tonofilaments were produced after 2 days in culture and increased thereafter with culture time, becoming condensed with desmosomes. Keratohyaline granules were observed in 7-day cultures. These keratinization characteristics occurring during culture showed the general characteristics of the alpha stratum observed in the skin of in ovo embryos during the early stages of development. However, the development of peridermal and subperidermal granules was poor and the stratum granulosum, which develops at the late stages between the stratum intermedium and the stratum corneum, was not observed. Polyacrylamide gel electrophoresis of S-carboxymethylated keratin proteins showed that the keratin protein band patterns of the culture differed from those of in ovo skin epidermis.