Pituitary luteinizing hormone and prolactin messenger ribonucleic acid levels are inversely related in laying and incubating turkey hens.

The relationships of prolactin (PRL) and LH messenger (m) RNA to serum and pituitary content were determined for turkey hens at different phases of the reproductive cycle. In the nonphotostimulated, reproductively inactive hen, serum and pituitary PRL content and pituitary PRL mRNA levels were low. All three PRL values rose after photostimulation and peaked during the incubation phase. Relative to nonphotostimulated hens, hyperprolactinemic incubating hens showed 220-, 11-, and 57-fold increases in serum PRL, pituitary PRL content, and pituitary PRL mRNA levels, respectively. These peak levels declined 80-, 3-, and 6-fold, respectively, in photorefractory hens. In contrast to PRL levels, serum LH, pituitary LH, and pituitary LH beta-subunit mRNA levels did not change as dramatically. Serum LH showed no significant changes for the different reproductive phases. Pituitary LH peaked after photostimulation and declined to its lowest level in incubating hens. Pituitary LH-beta mRNA abundance was highest in photostimulated and laying hens and lowest in incubating and photorefractory hens. These results demonstrate that the abundance of LH-beta and PRL mRNA shows an inverse relationship in photostimulated/laying and incubating turkey hens.     

Ovarian steroid production in vitro during gonadal regression in the turkey. I. Changes associated with incubation behavior.

The mechanism regulating ovarian regression during incubation behavior in the domestic turkey has not been elucidated. This study was designed to determine whether ovarian steroidogenic potential is depressed during gonadal regression associated with the onset of incubation behavior. Hens were housed in floor pens equipped with trap nests that were checked 7 times per day. Hens were grouped, according to nesting frequency and egg production, into the following classifications: laying (laid an egg every day and trapped in the nest only once/day); transitional (laid an egg every day but trapped in the nest 4 or more times/day); and Day 1, Day 3, and Day 5 incubating (no egg for 2, 4, or 6 days, respectively, while trapped in the nest at least 4 times/day). Follicular atresia was evident in the largest preovulatory follicle (F1) in transitional hens, extensive in F1 through the third largest follicle (F3) in Day 1 incubating hens, and extensive in F1 through F7in Day 3 incubating hens. Levels of circulating LH, progesterone (P), androgen (A), and estradiol (E) decreased in transitional hens relative to concentrations in laying hens and remained low thereafter. In contrast, levels of prolactin were greater in Day 3 and Day 5 incubating hens than in laying, transitional, or Day 1 incubating hens. Basal production of P by F1 granulosa cells was lower from Day 1 incubating hens than from the other groups. Production of P in response to porcine-luteinizing hormone (pLH) was greater by cells from transitional and Day 1 incubating hens than from those of laying hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma prolactin levels in incubating turkey hens during pipping of the eggs and after introduction of poults into the nest

Plasma levels of prolactin (Prl) associated with incubation and maternal behavior were compared in turkey hens allowed to incubate 10 fertile eggs (Group I, n = 9) or 10 infertile eggs (Group II, n = 7) in open nest boxes. At the end of the day that the first egg hatched, all unhatched eggs were removed from Group I hens and each hen was given 10 poults. At the end of the following day, infertile eggs were removed from Group II hens and each hen was given 10 poults. Although pipping of the eggs changed the incubation behavior of Group I hens, it had no effect on plasma Prl. Subsequent hatch of the eggs and/or presence of poults resulted, within 24 h, in a sharp fall in Prl levels, abandonment of the nests, and a shift to maternal behavior. Visual and auditory exposure to Group I poults had no effect on plasma Prl or incubation behavior of Group II hens incubating infertile eggs in adjacent pens. However, within 24 h after the infertile eggs were exchanged for newly hatched poults, Prl levels in Group II hens declined sharply and the hens abandoned the nests and showed maternal behavior similar to that observed in Group I hens. No significant relationships were found in either group between plasma Prl levels and quality of incubation or maternal behavior.     

Non-activated progesterone receptor extracted from nuclei of hen oviduct

When the progesterone receptor was extracted from nuclei of laying hen oviduct with 0.5 M sodium molybdate, a large, 7-8 S, form of the receptor was observed. This receptor form resembled non-activated cytoplasmic receptor not only in displaying the same sedimentation coefficient, but also in rapid dissociation rate of the hormone-receptor complex. This finding suggests that either activation may occur within the nuclear compartment, or that activation may be reversed under certain conditions.     

Immunoelectron microscopical demonstration of endogenous avidin in secretory granules of the hen oviduct mucosa: a preliminary study

Summary The localization of avidin in the oviduct of the laying hen was investigated using ultrastructural immunoperoxidase techniques. Endogenous avidin was localized in secretory granules of both tubular gland cells and non-ciliated single epithelial cells in the magnum mucosa. These immunospecific granules were electron-dense and heterogeneous with a patchy core and dense peripheral region, especially in acinar cells. The size varied from small to large in the gland cells (500–2200 nm in diameter) and remained small in the epithelial cells (180–720 nm). Columnar epithelial cells containing avidin granules strongly resembled the protodifferentiated tubular gland cells appearing in the magnum mucosa of chicks artificially pretreated with ovarian hormones. On the other hand, no avidin was observed in either epithelial goblet cells or ciliated cells in adult hens, although both cell types were shown to produce avidin in young chicks when synchronized by the administration of progesterone. The present results parallel those obtained with biotinylated enzyme affinity methods in our previous cytochemical study.Therefore, avidin is one of the proteins produced and stored in the secretory granules of the tubular gland cells and protodifferentiated acinar cells present in the epithelial layer of the laying hen oviduct. It is not present in goblet cells. Although the initiation of a synthesis may be triggered by progesterone, it is still not clear whether different hormone dependent proteins are localized in the same granules in both the adult hen and the immature chick.     

Effect of feed and water deprivation or force-feeding on plasma prolactin concentration in turkey hens

Plasma concentrations of prolactin (Prl), glucose, corticosterone, and D(-)-3-hydroxybutyrate (DBHB) were compared in nonlaying, nonincubating turkey hens subjected to feed and/or water deprivation. Neither Prl nor corticosterone concentrations were significantly (P greater than 0.05) altered by any of the treatments, whereas fasting significantly (P less than 0.05) reduced the concentration of glucose and increased the concentration of DBHB. Plasma levels of Prl in incubating hens were significantly (P less than 0.05) reduced by nest deprivation either in the absence of feed and water or when the hens were force-fed the normal intake for a laying hen. After 48 h of nest deprivation, the hens resumed nesting within 5 min of being returned to the pen although the plasma levels of Prl were low. Neither nest attentiveness nor the concentration of Prl were affected by force-feeding the hens while they were incubating eggs. The concentration of glucose increased in response to force-feeding or nest deprivation, whereas the concentration of corticosterone was increased only by force-feeding. These results suggest that Prl may not be involved in the striking changes in both intermediary and water metabolism which occur during incubation in the turkey hen. Furthermore, since incubation behavior can occur in the presence of low concentrations of Prl, elevated levels of Prl during broodiness appear to be maintained by a stimulus associated with the nest itself or some other aspect(s) of the environment.     

Adrenergic activity of the avian oviduct in vivo

The effect of noradrenaline or isoproterenol, alone or in the presence of an alpha (phenoxybenzamine) or beta (propranolol) adrenergic blocking drugs on the oviducts of anesthetized laying hens was investigated. The results show that both alpha and beta adrenergic activity is present in the avian oviduct with the exception of the uterus which does not appear to have alpha excitatory activity. Norepinephrine induced a strong contraction followed by a brief relaxation period in the infundibulum, magnum and is thmus; administration of phenoxybenzamine blocked this response in all the three segments, indicating the presence of alpha adrenergic receptors. The uterus, however, exhibited an inhibitory response in the majority of the hens and this response was not affected by the administration of phenoxybenzamine. Isoproterenol always induced relaxation in all the four segments of the oviduct. This response was blocked by propranolol, a beta adrenergic blocker, indicating the presence of beta adrenergic receptors. The role of autonomic nerves innervating the reproductive tract in the regulation of reproduction is discussed.     

Immunohistochemical localization of a calcium pump and calbindin-D28k in the oviduct of the laying hen.

The localization of a plasma membrane calcium pump in the oviduct of the laying hen was investigated by immunohistochemical techniques, utilizing a monoclonal antibody (5F10) produced against the human erythrocyte calcium pump. This antibody was shown to react with an epitope of the pump in oviductal tissue, and prominent staining was observed on the microvilli of the tubular gland cells of the hen shell gland (uterus) and the isthmus. The Ca2+ pump was not detectable in the infundibulum or the magnum. Calbindin-D28k, also localized by immunohistochemical means, was observed to be present in the tubular gland cells of the shell gland and the distal isthmus (adjacent to shell gland) but not in either the proximal isthmus (adjacent to the magnum), the magnum or the infundibulum. The localization of the Ca2+ pump in the oviduct corresponds to known sites of mineral deposition during egg shell formation. The distribution of calbindin-D28k differed, co-localizing with the Ca2+ pump in the shell gland and distal isthmus but not in the proximal isthmus. This might reflect a greater rate of active Ca2+ secretion in the distal isthmus and shell gland as compared to the proximal isthmus.     

Genotype analyses of Campylobacter isolated from distinct segments of the reproductive tracts of broiler breeder hens

Campylobacter isolated from feces and from the oviduct of six broiler breeder hens were genotyped by using flaA SVR DNA sequence analyses. A diversity of genotypes was observed among fecal and oviduct isolates. Comparison of isolates from the oviducts of individual hens revealed variable results. In three cases (hen 2, hen 3, and hen 6), analyses indicated that isolates from all regions of the individual hen's reproductive tract were closely related; isolates from hen 1 and hen 4 were diverse. Comparison of the Campylobacter isolates between hens revealed that in two cases, hens 1 and 3 and hens 4 and 6, certain isolates possessed identical flaA SVR sequence types. Comparisons of Campylobacter isolates recovered from a distinct region of the oviduct were found to have increased diversity as sampling progressed down the oviduct. This study further demonstrates that Campylobacter is present within the reproductive tract of breeder hens and that this presence may enable vertical transmission of Campylobacter from the breeder hen to the broiler offspring.     

Ultrastructural changes in the oviduct of the laying hen during the laying cycle

The ultrastructural changes occurring in the fully functional oviduct of Isa Brown laying hens were studied during various stages of the laying cycle. Hens were killed at different positions of the egg in the oviduct. The oviduct was lined by ciliated and non-ciliated cells (also referred to as granular cells). The granular cells in the infundibulum contributed to secretion during egg formation, whereas ciliated cells showed little evidence of secretion. Ultrastructural changes were recorded in the granular and glandular cells of the distal infundibulum. In the magnum, the surface ultrastructure revealed glandular openings associated with the ciliated and granular cells. Cyclic changes were recorded in the glandular cells of the magnum. With respect to the three observed types of glands, the structure of gland type A and C cells varied at different egg positions in the oviduct, whereas type B cells represented a different type of gland cell containing amorphous secretory granules. The surface epithelium of the isthmus was also lined by mitochondrial cells. Two types of glandular cell (types 1 and 2) were recorded in the isthmus during the laying cycle. Intracisternal granules were found in type 2 cells of the isthmus. A predominance of glycogen particles occurred in the tubular shell gland. The granular cells in the shell gland contain many vacuoles. During egg formation, these vacuoles regressed following the formation of extensive rough endoplasmic reticulum; the reverse also occurred. The disintegrated material found in the vacuoles may have been derived from the disintegrating granules. The Physiology Teaching Unit, University of New England, provided financial support to K. Chousalkar for this study.     

Avian gonadotropin-releasing hormones I and II in brain and other tissues in turkey hens

1. Radioimmunoassays were developed for measuring avian gonadotropin-releasing hormones I and II (cGnRH I and II) in tissue extracts during the reproductive cycle. 2. Hypothalamic concentrations of cGnRH I and II were qualitatively similar being lowest in non-photostimulated hens, greater in laying hens and greatest in incubating hens. 3. cGnRH II concentrations were similar in paraolfactory lobe and hypothalamic fragments while lesser amounts were found in cerebrum, cerebellum, duodenum, shell gland, and pineal. 4. These results suggest that cGnRH II has unknown functions in turkeys quite distinct from traditional functions associated with GnRHs.     

Changes in ovalbumin and protein synthesis in vivo in the magnum of laying hens during the egg formation cycle.

1. The present study was carried out to investigate whether or not the rate of synthesis of total protein in various oviducal segments and ovalbumin, a major egg white protein, in the magnum fluctuated during the egg formation cycle in laying hens. 2. Synthesis of total protein and ovalbumin was measured in vivo by the incorporation of [15N]methionine after a primed continuous infusion of tracer for 3 hr. 3. Protein and ovalbumin contents in the magnum and the entire oviduct decreased sharply when an ovum moved down from the magnum to the isthmus, probably due to the secretion of egg white proteins. 4. In contrast, total protein and ovalbumin synthesis in the magnum was significantly higher when an ovum was in there than when it was in any other segments. Fluctuations of ovalbumin synthesis and total protein synthesis in the magnum were roughly parallel to those of total protein synthesis in the entire oviduct. 5. It was concluded, therefore, that the changes seen in total protein synthesis in the whole oviduct during the egg formation cycle were mainly attributable to those in magnum protein synthesis, of which a significant portion was accounted for by the synthesis of ovalbumin.     

Effect of protein starvation on protein turnover in liver, oviduct and whole body of laying hens

1. Whole body protein turnover rates in White Leghorn laying hens were reduced by protein starvation for 7 days, followed by complete restoration by protein repletion for 7 days. 2. Protein starvation considerably reduced fractional and absolute rates of protein synthesis both in the liver and, to a greater extent, in the oviduct. 3. It was suggested that a considerable portion of the reduced whole body protein synthesis could be accounted for by the reduced protein synthesis in these organs when laying hens were subjected to protein starvation.     

Expression of vasoactive intestinal peptide receptor messenger RNA in the hypothalamus and pituitary throughout the turkey reproductive cycle

Vasoactive intestinal peptide (VIP) has been implicated in the regulation of avian reproductive activity and appears to act at the level of the hypothalamus and pituitary. This in situ hybridization histochemistry study describes the distribution of VIP receptor mRNA expression in the hypothalamus and the pituitary of reproductively active (laying) and quiescent (nonphotostimulated, incubating, and photorefractory) female turkeys and characterizes the differences observed in VIP receptor gene expression. VIP receptor mRNA, while expressed throughout the hypothalamus, was specifically expressed in areas known to contain GnRH-I neurons in the chicken, i.e., the lateral septum, medial preoptic area, anterior hypothalamus, and paraventricular nucleus. Significant differences in VIP receptor mRNA expression between different reproductive states was observed only within the infundibular nuclear complex. VIP receptor mRNA was markedly less in nonphotostimulated and photorefractory hens as compared with laying and incubating hens. The most dense VIP receptor mRNA was found in the anterior pituitary, where it was 2.4- and 3.0-fold greater in laying and incubating hens, respectively, as compared with that in nonphotostimulated ones. Hens that stopped incubating and became photorefractory displayed pituitary VIP receptor mRNA levels similar to those of nonphotostimulated birds. The changes in pituitary VIP receptor mRNA expression were positively correlated with known changes in pituitary prolactin (PRL) mRNA expression and PRL content and release. These findings indicate that the variations in PRL secretion observed across the turkey reproductive cycle are, in part, regulated by changes in VIP receptors at the pituitary level.     

Protection of steroid hormone receptors by protease inhibitors

Steroid hormone receptors are proteolyzed by different types of enzymes present in target tissues. Effective protease inhibitors protecting steroid hormone receptors in various target tissues were investigated. Progesterone receptor (PR) in hen oviduct and estrogen receptor (ER) in cow uterus were specifically protected by relatively low concentrations (0.5 mM) of leupeptin or antipain (inhibitors of serine and thiol proteases). It was indicated that two different types of enzymes which modify native glucocorticoid receptor (GR) are present in rat liver. One was inhibited by 1 mM leupeptin or 1 mM antipain, while the other was inhibited by 1 mM phosphoramidon (inhibitor of thermolysin like proteases) or 10 mM sodium molybdate. Native PR, ER, and GR were shown to have similar Stokes radii (44 Å).     

Effects of early prenatal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on postnatal reproduction in the laying hen (Gallus gallus)

This study demonstrates the long-term effects of very early embryonic exposure to a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (0, 10 and 20 ng/egg), administered before the beginning of embryonic development, on growth and reproductive performance in laying hens. Hatchability and body weight gain from 11 weeks onwards were significantly depressed in 20 ng treated hens. All hens started laying egg at around the same age and the laying performance of TCDD-treated hens was normal. No disturbances in the age-related pattern and concentrations of oestradiol, LH or FSH in plasma could be found but mean progesterone concentrations were significantly lower in 20 ng treated hens. Moreover, follicular distribution was changed with less small white follicles and smaller yellow follicles, which probably resulted in the lower egg weight of the 20 ng treated hens. At 43 weeks of age, hens treated in ovo with TCDD showed a retained right oviduct, mostly filled with clear fluid. From these results, it seems that in ovo exposure to TCDD interferes in the right oviduct regression during embryonic development and induces some changes in follicular distribution but without impairment of reproductive performance in the adult laying hen.     

Oestrogen receptors in chick oviduct. Characterization and subcellular distribution.

Macromolecular components with properties of oestrogen receptors have been identified in the 0.5 M KCl nuclear soluble, the nuclear insoluble and the cytosol fractions of laying hen and immature (2--4 weeks, untreated by hormone) chicken oviduct. 7n the 0.5 M KCl extract of laying hen oviduct nuclei, a receptor, of protein nature according to the effects of enzymic treatments, has been identified. It exhibits high affinity for oestradiol with an apparent equilibrium association constant KA = 4 - 109 M-1 at 4 degrees C. The binding of [3H] oestradiol is abolished by 1 muM oestriol, oestrone and diethylstilboestrol, but not by the same concentration of progesterone, testosterone, and cortisol. Sucrose gradient ultracentrifugation studies in the presence of 0.5 M KCl indicate a sedimentation coefficient of 4.3 S, and there is partial aggregation in low-ionic-strength medium. The estimated number of binding sites per nucleus is about 5000, as calculated from DNA content of chick diploid genome. Most of the binding sites were found to be occupied by endogenous oestrogen(s). Oestradiol dissociates from the receptor according to an apparent two-step mechanism. The half-life time for the faster dissociation step is 18 h at 0 degrees C, 25 min at 20 degrees C and 10 min at 30 degrees C, and for the slower one is 180 h, 115 min and 60 min, respectively. In the 0.5 M KCl extract of immature chicken oviduct nuclei, there are approximately 500 receptor sites per nucleus; their affinity for oestradiol is the same as in the case of laying hen soluble nuclear receptor. After repeated extractions of nuclei with 0.5 M KCl medium, a substantial quantity of oestrogen binding sites remains in the residual fraction. Binding characteristics of this insoluble nuclear receptor resemble those of the soluble nuclear receptor: high affinity for oestradiol (KA = 7 - 10(8) M-1 at 37 degrees C) and specificity for oestrogens. The estimated number of binding sites are approximately 2000/cell for laying hen, and approximately 1000/cell for immature chicken. In the high-speed supernatant fraction of laying hen oviduct homogenates, an oestrogen receptor is also present, but its concentration is low (less than or equal to 100 sites/cell) and at the limits of sensitivity of the methods used. In the cytosol of immature chicken oviduct, there are approximately 2500 oestradiol receptor sites per cell.     

Genotype Analyses of Campylobacter Isolated from Distinct Segments of the Reproductive Tracts of Broiler Breeder Hens

Campylobacter isolated from feces and from the oviduct of six broiler breeder hens were genotyped by using flaA SVR DNA sequence analyses. A diversity of genotypes was observed among fecal and oviduct isolates. Comparison of isolates from the oviducts of individual hens revealed variable results. In three cases (hen 2, hen 3, and hen 6), analyses indicated that isolates from all regions of the individual hen's reproductive tract were closely related; isolates from hen 1 and hen 4 were diverse. Comparison of the Campylobacter isolates between hens revealed that in two cases, hens 1 and 3 and hens 4 and 6, certain isolates possessed identical flaA SVR sequence types. Comparisons of Campylobacter isolates recovered from a distinct region of the oviduct were found to have increased diversity as sampling progressed down the oviduct. This study further demonstrates that Campylobacter is present within the reproductive tract of breeder hens and that this presence may enable vertical transmission of Campylobacter from the breeder hen to the broiler offspring. Received: 11 January 2002 / Accepted: 13 March 2002     

Immunohistochemical localization of a calcium pump and calbindin-D28k in the oviduct of the laying hen

Summary The localization of a plasma membrane calcium pump in the oviduct of the laying hen was investigated by immunohistochemical techniques, utilizing a monoclonal antibody (5F10) produced against the human erythrocyte calcium pump. This antibody was shown to react with an epitope of the pump in oviductal tissue, and prominent staining was observed on the microvilli of the tubular gland cells of the hen shell gland (uterus) and the isthmus. The Ca²⁺ pump was not detectable in the infundibulum or the magnum. Calbindin-D_28k, also localized by immunohistochemical means, was observed to be present in the tubular gland cells of the shell gland and the distal isthmus (adjacent to shell gland) but not in either the proximal isthmus (adjacent to the magnum), the magnum or the infundibulum. The localization of the Ca²⁺ pump in the oviduct corresponds to known sites of mineral deposition during egg shell formation. The distribution of calbindin-D_28k differed, co-localizing with the Ca²⁺ pump in the shell gland and distal isthmus but not in the proximal isthmus. This might reflect a greater rate of active Ca²⁺ secretion in the distal isthmus and shell gland as compared to the proximal isthmus.