Comparison of plasma ACTH and corticosterone levels after embryo bursectomy

Plasma levels of both adrenocorticotropic hormone (ACTH) and corticosterone (B) were determined in embryos (day 15 of incubation), chicks (day 3 after hatch) and young chickens (8 weeks). Experimental animals were bursectomized at 80 hr of incubation, i.e., before any anlage of the bursa of Fabricius could develop. Bursectomized (BFX) animals were compared to sham-operated controls (T), in basal, resting condition and 7 (ACTH) or 14 min (B) after ether stress was delivered for 30 sec. Basal B and ACTH levels seemed not to be significantly modified in BFX embryos, chicks and chickens. Hypophysial and adrenocortical response to stress appeared more precociously in BFX embryos (day 15 of incubation) than in intact ones (day 19). The non stress-responsive period that was observed for one week after hatch of T birds did not appear in 3-day-old BFX chicks whose both B and ACTH stress-induced levels were as high as in intact adults. In contrast, adrenocortical and pituitary corticotropic responses to stress were markedly impaired (by 50%) in adult BFX chickens as compared to intact controls.     

Effect of ACTH injection and graft of bursa of Fabricius on the adrenocortical activity of bursectomized chicks at 80 hours of incubation

Early embryonic bursectomy (BFX) disturbed the adrenocortical functioning. The stress-unresponsive period that occurred in controls, and lasted for 2-3 weeks after hatching, no longer appeared in BFX chicks. In contrast, the magnitude of the stress-induced hypercorticosteronemia was much lower in BFX than in sham-operated 5 week-old chicken. It was assumed that such adrenocortical dysfunction was due to bursal deprivation, since grafting bursal buds onto the chorio-allantoic membrane of BFX embryos restored all the parameters under study, i.e., the post-hatching stress unresponsive period and the high magnitude of stress-induced responses in adults. Factor(s) involved in such interregulation are not known but do not seem to affect directly adrenocortical cells because intramuscular injection of a moderate dose of ACTH resulted in the same hypercorticosteronemia whether 3 day-old and 5 week-old chicks had been bursectomized or sham-operated.     

Developmental changes in vitamin A level and lack of retinyl palmitate in chick lungs

1. Developmental changes in retinol and retinyl palmitate contents in lungs of chick embryos and posthatch chicks were investigated. 2. Remarkable changes in the lung retinol levels were found during development of chicks. Embryonic lungs 5 days prior to hatching contained the highest content of retinol. The level then declined rapidly and was lowest on 1 day before hatching. 3. Its level then rose substantially within 7 days after hatching. 4. No retinyl palmitate in chick lungs was detectable at any of the developmental stages examined, nor even in adult hen. 5. Serum retinol level changed in parallel with the lung retinol. 6. The patterns of changes in liver retinol and retinyl palmitate were remarkably different from that occurring in the lung retinol. In chick embryonic livers, the levels of them were low, followed by a rapid increase after hatching. 7. The high level and its rapid decrease of lung retinol content during development of chick embryos may be functionally connected with retinol action in embryonic lungs for cellular differentiation and maturation.     

A hormonal misunderstanding in Acca sellowiana embryogenesis: levels of zygotic embryogenesis do not match those of somatic embryogenesis

Endogenous levels of IAA, ABA and four types of CKs were analyzed in zygotic and indirect (ISE) and direct somatic embryogenesis of Acca sellowiana. Zygotic and somatic embryos at different developmental stages were sampled for morphological and hormonal analysis. Both embryo types showed substantial asymmetry in hormone levels. Zygotic embryos displayed a conspicuous peak of IAA in early developmental stages. The results outlined the hormonal variations occurring during zygotic and somatic embryogenesis regarding the timing, nature and hormonal status involved in both processes. The short transient pulse of IAA observed on the 3rd day in culture was suggested to be involved with the signaling for the induction of somatic embryogenesis. Fertilized ovule development was associated with increased IAA levels 21?C24?days after pollination, followed by a sharp decrease in the cotyledonary stage, both in zygotic and somatic embryos. There was a prominent increase in ABA levels in cultures which generated ISE 24?C30?days after pollination, a period that corresponds to the heart and torpedo stages. The levels of total CKs (Z, [9R]Z, iP and [9R]iP) were also always higher in zygotic than in somatic embryogenesis. While zygotic embryogenesis was dominated by the presence of zeatin, the somatic process, contrarily, was characterized by a large variation of the other cytokinin forms and amounts studied. The above results, when taken together, could be related to the previously observed high frequency formation of anomalous somatic embryos formed in A. sellowiana, as well as to their low germination ability.     

Development of blood testosterone in the embryo and young chick after precocious bursectomy

The effect of bursa of Fabricius on the endocrine function of the chick testes was studied in vivo by comparing plasma testosterone levels from 48 h before hatch to 16 weeks of age in both intact and bursectomized chicken. Early bursectomy was performed at 80 h of incubation. Post surgery survival was low (12% at 1 week). In controls, plasma testosterone levels were found to be low (100-200 pg. ml-1) from 48 h before to 48 h after hatch, then to raise up to a plateau (2,200 pg. ml-1) at 6 weeks. After bursectomy, values were first higher than in intact (210-440 pg. ml-1 from 48 h before/after hatch and 515 vs 300 pg.ml-1 at 3 days) but no difference could be further detected from 1 to 16 weeks of age. It is suggested that, in addition to the effect of androgen on bursa of Fabricius, the later reciprocally influences the gonadotropic axis during the early stage of development.     

The role of polarity in the development of the hydrozoan planula larva

Summary These experiments were done in order to define the role that polarity plays during embryogenesis in hydrozoans.Parts of hydrozoan embryos isolated at different developmental stages from early cleavage to postgastrula will regulate to form normal planulae. During this process, the original anterior-posterior axis of the part is conserved. In normal embryos the posterior pole of the anterior-posterior axis is congruent with the site where the polar bodies are given off and with the site where the first cleavage is initiated. By centrifuging fertilized eggs, it is possible to create embryos in which the first cleavage initiation site does not correspond to the site where the polar bodies are given off. In these embryos the posterior pole of the anterior-posterior axis corresponds to the first cleavage initiation site. When parts of these embryos are isolated at different stages they also regulate to form normal planulae. The axial properties of these planulae are determined by the site of first cleavage initiation.The interactions between regions of the embryo with different axial properties were studied by grafting together parts in such a way as to create embryos with abnormal axial arrangements. Following gastrulation interactions take place between the grafted parts leading to the formation of normal planulae with a new set of axial properties.Blastula stage embryos can be dissociated into single cells and the cells can be reaggregated. These reaggregates form normal planulae. Polarity can be entrained in the reaggregates by grafting a small piece of tissue from any part of an intact blastula to the reaggregate. These cells organize the formation of an axis of symmetry with an appropriate orientation with respect to the graft.     

Chick gonad differentiation following excision of primordial germ cells

The differentiation of embryonic chick gonads lacking germ cells was compared to that of normal chick gonads to determine whether the somatic elements of sterile avian gonads will undergo normal sexual differentiation. Primordial germ cells were removed by surgical excision of anterior germinal crescent from early embryos, Hamburger and Hamilton stages 6–11. Surgically treated and control embryos were sacrificed at 6, 15, and 20 days of incubation, and their gonads were studied histologically. Analysis of differentiation was based on morphological criteria at the cellular, tissue, and organ levels. In both male and female embryos, the somatic elements of the gonads differentiated normally in the absence of germ cells. The significance of these results for understanding the controls of differentiation of both the somatic gonad and the germ cells in birds is discussed and correlated with similar results in mammals.     

Effects of glucocorticoids on circulating concentrations of thyroxine (T4) and triiodothyronine (T3) and on peripheral monodeiodination in pre- and post-hatching chickens

The administration of either glucocorticoids (dexamethasone or corticosterone) or adrenocorticotropic hormone (ACTH) to chicken embryos was followed by increase in the circulating concentration of triiodothyronine (T3), the T3 to thyroxine (T4) ratio and the activity of liver T4-5' monodeiodinase. No consistent changes in plasma concentrations of T4 or GH were observed. In post-hatching chicks, corticosterone and dexamethasone depressed the circulating concentrations of both T4 and T3. Iopanoc acid, an inhibitor of liver T4-5' monodeiodinase, elevated plasma concentrations of T4 and depressed those of T3 in both chicken embryos and young chicks. It is suggested that glucocorticoids affect circulating concentrations of T4 and T3 both by affecting the activity of the liver T4-5' monodeiodinase and by influencing the hypothalamo-pituitary axis.     

Development of spinal cord bioelectric activity in spinal chick embryos and its behavioral implications

Embryonic behavior of the chick is the product of spontaneous multiunit burst discharges within the ventral spinal cord. The present study describes the ontogeny of spinal cord burst discharges in embryos which were deprived of brain input by removing several neural tube segments of 2-day embryos at cervical or mid-thoracic levels. Characteristics of bioelectric activity present in both intact and chronically transected cords are: (a) the appearance of spike discharges; (b) the organization of unit discharges into synchronized multiunit bursts; (c) the establishment of intracord synchronization of burst discharges over wide expanses of cord tissue; (d) an increase in burst duration and complexity at 7 days due to the appearance of the burst afterdischarge; (e) an increase in the amount of burst activity from 6 to 13 days followed by a decline until hatching at 21 days; (f) a shift from periodic to irregular patterns of burst activity at 13 days; and (g) the existence of the cord burst discharge as a correlate of embryonic movement. Several differences were found between burst activity from chronic spinal and intact embryos: (a) cervical spinal embryos were significantly less active than controls from 15 through 19 days; and (b) long sequences of unusual repetitive burst afterdischarges appeared in chronic spinal embryos by 13 days. The results indicate that the transected embryonic spinal cord is remarkably autogenous in function, although patterns of activity unique to the transected cord appear and increase in prominence during later stages of incubation.     

Creatine phosphokinase activity in dysgenic (mdg/mdg) mouse muscle

The specific activity of creatine phosphokinase (CPK) was measured in the muscle of mdg/mdg and control embryos of 14-18 days' gestation. CPK specific activity values were similar in mutant and normal embryos at the earliest stages examined (14-15 days). However, after 15 1/2 days, the specific activity of the enzyme in the mdg/mdg embryos was approximately 50% lower than in the controls. The dysgenic and normal muscle extracts exhibited comparable stability after storage at -85 C. CPK activity levels in the muscle of adult heterozygotes (+/mdg) and wild-type (+/+) controls were found to be statistically identical. The findings suggest that the mdg mutation does not have a primary or direct effect on CPK activity.     

Regulation and the modification of axial properties in partial embryos of the nemertean, Cerebratulus lacteus

 Embryos acquire axial properties (e.g., the animal-vegetal, dorsoventral and bilateral axes) at various times over the course of their normal developmental programs. In the spiral-cleaving nemertean, Cerebratulus lacteus, lineage tracing studies have shown that the dorsoventral axis is set up prior to the first cleavage division; however, blastomeres isolated at the two-cell stage will regulate to form apparently perfect, miniature pilidium larvae. We have examined the nature of axial specification in this organism by determining whether partial embryos retain the original embryonic/larval axial properties of the intact embryo, or whether new axial relationships are generated as a consequence of the regulatory process. Single blastomeres in two-cell stage embryos were injected with lineage tracer, and were then bisected along the second cleavage plane at the four-cell stage. Thus, the relationship between the plane of the first cleavage division and various developmental axes could be followed throughout development in the ”half-embryos”. While some embryo fragments appear to retain their original animal-vegetal and dorsoventral axes, many fragments generate novel axial properties. These results indicate that axial properties set up and used during normal development in C. lacteus can be completely reorganized during the course of regulation. While certain embryonic axes, such as the animal-vegetal and dorsoventral axes, appear to be set up prior to first cleavage, these axes and associated cell fates are not irreversibly fixed until later stages of development in normal intact embryos. In C. lacteus, the process whereby these properties are ultimately determined is apparently controlled by complex sets of cell-cell interactions. Received: 11 October 1996 / Accepted: 21 February 1997     

Thyroid hormone response to thyrotropin releasing hormone after cold treatment during pre- and postnatal development in the domestic fowl

The purpose of the present study was to compare the effect of periodic cooling during the establishment of a functional pituitary-thyroid axis at days 11-14 of incubation and at other developmental stages, on the subsequent thyroid hormone response to thyrotropin releasing hormone (TRH). In the first and second experiment chick embryos were cooled for 6 hr/day to 30 degrees C from day 11 till 14 and from day 15 till 18 respectively, whereas control groups were incubated throughout at 37.8 degrees C. In both experiments the thyroxine (T4) response upon TRH in 19 day-old embryos was higher in the previously cold treated embryos, according to the percentages of increase. However, the higher T4 response in the cold treated animals disappeared in 1 or 7 day-old chicks hatched from the 2nd experiment, but remained present in chicks of the same ages in the 1st experiment. In a third experiment the T4 response to TRH injection immediately and 3 and 8 days after a temperature treatment (25 degrees C or 12 degrees C) for one week on four weeks old broiler chickens was found to be similar in both temperature groups. In all experiments there was a concomitant triiodothyronine (T3) increase after TRH injection, but differences between experimental groups were observed at days 15 and 19 of incubation and immediately after the postnatal temperature treatment. As an overall conclusion the results indicate that cold treatment only during the establishment of the hypothalamo-hypophysial control of thyroid function can have a long lasting effect by enhancing the T4 response to TRH injection.     

The adrenocortical stress-response of Black-legged Kittiwake chicks in relation to dietary restrictions

In this study we examined hormonal responses of Black-legged Kittiwake (Rissatridactyla) chicks to experimental variations in energy content and nutritional quality (low or high lipid to protein ratio, LPR) of their food. Starting at the age of 10 days, chicks were fed either high or low LPR fish at 30, 50, 70 and 100% of ad libitum energy intake. After 20 days of treatment, chicks were exposed to a standardized acute handling and restraint stress protocol, where a baseline sample was taken immediately after taking a chick from the nest, and three additional blood samples were taken at intervals up to 50 min. Testosterone and corticosterone titres in plasma were measured via radioimmunoassay. We found that baseline testosterone levels were not significantly affected by the experimental treatments. Food-restricted chicks had elevated baseline and acute stress-induced levels of corticosterone compared to chicks fed ad libitum. An elevation of circulating levels of corticosterone in energetically stressed individuals was further magnified by low nutritional quality of food. Baseline and acute stress-induced corticosterone levels of chicks were negatively correlated with their fat reserves. We conclude that the physiological condition of Black-legged Kittiwake chicks can be assessed reliably by measuring circulating levels of corticosterone. We discuss short- and long-term effects of elevated corticosterone secretion in food-stressed nest-bound chicks. Accepted: 13 April 1999     

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.     

Hormonal regulation of fatty acid synthetase in chick embryo liver.

Fatty acid synthetase activity in chick embryonic liver is negligible compared to that in newly hatched, fed chicks. The enzyme activity is prematurely induced 5–50-fold in 20-day-old embryos and in newly hatched chicks by the administration of insulin, hydrocortisone, growth hormone, glucagon or dibutyryl cyclic AMP. The induction of the enzyme activity is blocked by the administration of cycloheximide, indicating that new protein synthesis is required. Immunochemical titrations of different enzyme preparations from 5-day-old chicks, adult chicken and various inducer-treated embryos gave an identical equivalence point, indicating that the changes in synthetase activity after hormonal induction in embryos are related entirely to changes in content of enzyme. The increase in liver synthetase content after administration of insulin, glucagon or dibutyryl cyclic AMP is directly related to an increase in the rate of synthetase synthesis. The induction of the synthetase activity by suboptimal doses of glucagon or cyclic AMP is potentiated by the phosphodiesterase inhibitory theophylline. There is a very rapid decay of synthetase activity, with a half-life of about 4 h after elevation to higher levels following administration of insulin, glucagon or dibutyryl cyclic AMP. Glucagon and dibutyryl cyclic AMP induction of the synthetase activity is observed early in the embryonic development, whereas insulin induction is noted 2 days before hatching. Insulin, glucagon and cyclic AMP are potentially capable of altering the levels of glycolytic intermediates which may be involved in the induction of synthetase.     

4-14C-progesterone conversion by the fetal rat adrenal gland in vitro.

The adrenal glands of rat fetuses with activated or inhibited pituitary adrenocorticotropic activity between the 15th and 22nd day of intrauterine development were incubated with 4-14C-progesterone for 3hr. Fetuses of intact mothers were used as controls. Conversion of progesterone into adrenal steroids was found increased on the 18th day of intrauterine development, i.e., at the time when fetal adrenocorticotropic activity begins. In comparison to controls, conversion of progesterone into fetal adrenal corticosteroids was the smallest in the fetuses of mothers with inhibited pituitary ACTH and the greatest in the adrenals of fetuses of mothers with activated pituitary adrenocorticotropic activity.     

Activation of the serotonergic system in chick spinal cord during hatching.

The objectives of the present study were to determine the levels of serotonin (5-HT), its major catabolic metabolite, 5-hydroxyindoleacetic acid (5-HIAA), and norepinephrine (NE) in chick spinal cord before, during, and after hatching and also to determine if changes in the levels of these chemicals are directly related to the hatching behavior. The levels of 5-HT, 5-HIAA, and NE were measured by high performance liquid chromatography with electrochemical detection in whole spinal cords of 20-day-old "pre-hatching" embryos, 21-day-old "normal hatching" embryos, 0-day-old "post-hatching" chicks, and 0-day-old "glass egg hatching" chicks. NE was measured but no significant differences were found in NE levels among experimental groups. The concentration of 5-HT was elevated in chick embryo spinal cords during normal hatching compared to pre-hatching embryos and post-hatching chicks. The concentration of 5-HIAA was elevated during and after normal hatching compared to pre-hatching embryos. However, neither 5-HT nor 5-HIAA levels were found to be elevated in chick spinal cords during glass egg hatching compared to pre-hatching embryos or post-hatching chicks. Therefore, there appears to be an activation of the serotonergic system in chick spinal cord related to the specific event of hatching but this activation is not directly related to the movements common to both hatching and glass egg hatching.     

Hormonal basis of hysterectomy-induced maternal behavior during pregnancy in the rat.

Hysterectomy during the last half of pregnancy (i.e., Day 10–19) induces a rapid onset of maternal behavior; ovariectomy in addition to hysterectomy, prevents this effect. Estradiol and progesterone were tested for their ability to restore short-latency maternal behavior in hysterectomized-ovariectomized (HO) females operated on the 10th, 13th, 16th and 19th days of pregnancy. A single injection of either 20 μg/kg or 100 μg/kg estradiol benzoate (EB) immediately following HO either alone or followed by 0.5 mg progesterone (P) 44 hr later restored short-latency maternal behavior similar to that observed following hysterectomy only. The lower dose of EB was found to be equally effective at all stages of pregnancy and P was unnecessary to induce maternal behavior. The effectiveness of EB in inducing maternal behavior was discussed in relation to the hormonal changes which follow hysterectomy during pregnancy and to those which are associated with the normal onset of maternal behavior around parturition.     

ABA associated biochemical changes during somatic embryo development in Camellia sinensis (L.) O. Kuntze

The effect of ABA on reserve accumulation in maturing somatic embryos of tea was compared with and without ABA treatment. Changes in the levels of starch, total soluble sugars (TSS), proteins, and phenols were studied in the somatic embryos at different stages of development (globular, heart, torpedo and germinating embryos) in order to investigate whether ABA could trigger accumulation of storage reserves and thereby overcome the problem of poor germination. After ABA treatment (5.0 mg l(-1)) for 14 days, the starch and protein contents that were negligible in the untreated embryos increased by several fold with a simultaneous increase in TSS. When ABA treatment occurred at the heart stage, the germination of the embryos also improved, relative to untreated controls, after ABA treatment. ABA treatment prior to or after heart stage did not improve somatic embryo germination.