Project description:Measurement of blood corticosterone concentrations has been established as an indicator for assessment of acute distress. Therefore, knowledge on physiological fluctuations is required, but previous studies allow little conclusion on daily fluctuations in domestic chickens (Gallus gallus domesticus). To verify the presence of a circadian corticosterone rhythm in socialized chickens, blood samples were taken at four-hour intervals from 12 laying hens kept in groups of four over three days, each. Prior to experiments, hens were adapted to repeated handling for stress reduction. Corticosterone concentration was determined using radioimmunoassay. Blood sampling time and duration were recorded, and audio and video recordings were analyzed to assess the impact of behavior on corticosterone concentrations. Despite individual fluctuations, most hens showed a circadian course with two peaks per day. Statistics revealed a significant peak during the day (between 12:00 p.m. and 04:00 p.m.) and a tendency for a second peak at night (12:00 a.m.). The daily corticosterone peak was not explained by daytime social stress and needs to be seen as an endophenotype. The role of nightly corticosterone production has to be investigated in further studies. There might be a relation between corticosterone and reproduction since the only hen not showing peaks was not laying eggs.
Project description:BackgroundAlthough chicken oviduct is a useful model and target tissue for reproductive biology and transgenesis, little is known because of the highly specific hormonal regulation and the lack of fundamental researches, including lectin-binding activities and glycobiology. Because lectin is attached to secreted glycoproteins, we hypothesized that lectin could be bound to secretory egg-white proteins, and played a crucial role in the generation of egg-white protein in the oviduct. Hence, the purpose of this study was to investigate the structural, histological and lectin-binding characteristics of the chicken oviductal magnum from juvenile and adult hens.MethodsThe oviductal magnums from juvenile and adult hens were prepared for ultrastructural analysis, qRT-PCR and immunostaining. Immunohistochemistry of anti-ovalbumin, anti-ESR1 and anti-PGR, and mRNA expression of egg-white genes and steroid hormone receptor genes were evaluated. Lectin histochemical staining was also conducted in juvenile and adult oviductal magnum tissues.ResultsThe ultrastructural analysis showed that ciliated cells were rarely developed on luminal surface in juvenile magnum, but not tubular gland cells. In adult magnum, two types of epithelium and three types of tubular gland cells were observed. qRT-PCR analysis showed that egg-white genes were highly expressed in adult oviduct compared with the juvenile. However, mRNA expressions of ESR1 and PGR were considerably higher in juvenile oviduct than adult (P < 0.05). The immunohistochemical analysis showed that anti-ovalbumin antibody was detected in adult oviduct not in juvenile, unlikely anti-ESR1 and anti-PGR antibodies that were stained in both oviducts. In histological analysis, Toluidine blue was stained in juvenile and adult oviductal epithelia, and adult tubular glands located in the outer layer of oviductal magnum. In contrast, PAS was positive only in adult oviductal tubular gland. Lectins were selectively bound to oviductal epithelium, stroma, and tubular gland cells. Particularly, lectin-ConA and WGA were bound to electron-dense secretory granules in tubular gland.ConclusionsThe observation of ultrastructural analysis, mRNA expression, immunohistochemistry and lectin staining showed structural and physiological characterization of juvenile and adult oviductal magnum. Consequently, oviduct study could be helped to in vitro culture of chicken oviductal cells, to develop epithelial or tubular gland cell-specific markers, and to understand female reproductive biology and endocrinology.
Project description:Despite decades of investigation, the function of interphotoreceptor retinoid binding protein (IRBP), the most abundant protein in the interphotoreceptor matrix of vertebrates, remains enigmatic. Roles for IRBP in the visual cycle of rod photoreceptors and in the independent visual cycle of cone photoreceptors have been suggested, yet very little is known of the biology of IRBP in cone-dominant retinas, such as those of diurnal birds. Our aim was to identify and characterize expression of the IRBP of the cone-dominant chicken (Gallus gallus domesticus).Chicken IRBP mRNA was identified by PCR cloning. Primary protein structure, genomic organization, and phylogenies were determined through comparative sequence analyses. Expression of IRBP mRNA was characterized by northern analysis and by in situ hybridization on cryosectioned chicken retina. Expression of the IRBP protein was characterized by western blotting and by indirect immunofluorescence on cryosectioned retina and on retinal whole mounts.The chicken IRBP gene encodes a secreted protein with a predicted 1,252 amino acid length. The gene structure for chicken IRBP resembles that of most other vertebrates, with four homologous, modular repeats and introns within only the fourth module. Each module is more homologous with the corresponding module in other species than it is with the remaining chicken modules. Chicken retinal tissue contains a single IRBP mRNA transcript of approximately 4.8 kb and western analysis of chicken retina shows a single major band of 140 kDa. Chicken IRBP mRNA is expressed exclusively by retinal photoreceptor cells and the intensity of the hybridization signal shows light/dark rhythmicity. The IRBP protein is localized to the interphotoreceptor matrix of the chicken retina and to intracellular regions of photoreceptors, with a spatial distribution indicating an association with cone outer segments.The high degree of conservation of IRBP's primary structure, genomic organization, and cell-specific expression within the retinas of all vertebrates examined to date, including those with cone-dominant retinas, implies a conserved role for IRBP in photoreceptor function and/or health. Expression of chicken IRBP and its mRNA are functionally regulated. This report provides a necessary first step to explore a specific function for IRBP in the cone visual cycle.
Project description:Wild birds modulate wing and whole-body kinematics to adjust their flight patterns and trajectories when wing loading increases flight power requirements. Domestic chickens (Gallus gallus domesticus) in backyards and farms exhibit feather loss, naturally high wing loading, and limited flight capabilities. Yet, housing chickens in aviaries requires birds to navigate three-dimensional spaces to access resources. To understand the impact of feather loss on laying hens' flight capabilities, we symmetrically clipped the primary and secondary feathers before measuring wing and whole-body kinematics during descent from a 1.5 m platform. We expected birds to compensate for increased wing loading by increasing wingbeat frequency, amplitude and angular velocity. Otherwise, we expected to observe an increase in descent velocity and angle and an increase in vertical acceleration. Feather clipping had a significant effect on descent velocity, descent angle and horizontal acceleration. Half-clipped hens had lower descent velocity and angle than full-clipped hens, and unclipped hens had the highest horizontal acceleration. All hens landed with a velocity two to three times greater than in bird species that are adept fliers. Our results suggest that intact laying hens operate at the maximal power output supported by their anatomy and are at the limit of their ability to control flight trajectory.
Project description:The extracellular calcium-sensing receptor (CaSR) and vitamin D receptor (VDR) play important roles in regulating calcium mobilization, calcium absorption, and calcium homeostasis, and they could be potential therapeutic targets to osteoporosis in laying hens. The present study investigated the molecular distribution of CaSR and VDR and the localization of CaSR in the kidney, proventriculus (true stomach), duodenum, jejunum, ileum, colon, cecum, shell gland, and tibia of laying hens at 3 different laying stages (19, 40, and 55 wk). The results showed that the relative mRNA abundance of CaSR in the kidney, ileum, proventriculus, duodenum, and colon was higher (P < 0.05) than the other tissues at 40 and 55 wk. The relative mRNA abundance of CaSR in the tibia was higher (P < 0.05) at 55 wk than at 40 wk. However, there were no significant differences in the relative protein abundance of CaSR among all tested tissues at peak production or in each tissue at the 3 different laying stages (P > 0.05). The relative mRNA abundance of VDR was higher (P < 0.05) in the small intestine (duodenum, jejunum, and ileum) when compared with other tissues at the 3 different laying stages. The relative protein abundance of VDR in the duodenum was higher (P < 0.05) than that in the proventriculus, colon, and cecum. There were no significant differences in the VDR expression among the tested tissues at the 3 different laying stages (P > 0.05). The immunohistochemical results showed that the positive staining was found widely in each tissue. Moreover, different laying stages did not affect the localization of CaSR except for the tibia tissue. In conclusion, similar to VDR, CaSR was widely expressed not only in the gut but also in the tibia and shell gland in laying hens. The expression level of CaSR and VDR in all tested tissues was unchanged at the different laying stages.
Project description:BackgroundBroodiness significantly impacts poultry egg production, particularly notable in specific breeds such as the black-boned Silky, characterized by pronounced broodiness. An understanding of the alterations in ovarian signaling is essential for elucidating the mechanisms that influence broodiness. However, comparative research on the characteristics of long non-coding RNAs (lncRNAs) in the ovaries of broody chickens (BC) and high egg-laying chickens (GC) remains scant. In this investigation, we employed RNA sequencing to assess the ovarian transcriptomes, which include both lncRNAs and mRNAs, in eight Taihe Black-Bone Silky Fowls (TBsf), categorized into broody and high egg-laying groups. This study aims to provide a clearer understanding of the genetic underpinnings associated with broodiness and egg production.ResultsWe have identified a total of 16,444 mRNAs and 18,756 lncRNAs, of which 349 mRNAs and 651 lncRNAs exhibited significantly different expression (DE) between the BC and GC groups. Furthermore, we have identified the cis-regulated and trans-regulated target genes of differentially abundant lncRNA transcripts and have constructed an lncRNA-mRNA trans-regulated interaction network linked to ovarian follicle development. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analyses have revealed that DE mRNAs and the target genes of DE lncRNAs are associated with pathways including neuroactive ligand-receptor interaction, CCR6 chemokine receptor binding, G-protein coupled receptor binding, cytokine-cytokine receptor interaction, and ECM-receptor interaction.ConclusionOur research presents a comprehensive compilation of lncRNAs and mRNAs linked to ovarian development. Additionally, it establishes a predictive interaction network involving differentially abundant lncRNAs and differentially expressed genes (DEGs) within TBsf. This significantly contributes to our understanding of the intricate interactions between lncRNAs and genes governing brooding behavior.
Project description:Often referred to as the silent killer, ovarian cancer is the most lethal gynecologic malignancy. This disease rarely shows any physical symptoms until late stages and no known biomarkers are available for early detection. Because ovarian cancer is rarely detected early, the physiology behind the initiation, progression, treatment, and prevention of this disease remains largely unclear. Over the past 2 decades, the laying hen has emerged as a model that naturally develops epithelial ovarian cancer that is both pathologically and histologically similar to that of the human form of the disease. Different molecular signatures found in human ovarian cancer have also been identified in chicken ovarian cancer including increased CA125 and elevated E-cadherin expression, among others. Chemoprevention studies conducted in this model have shown that decreased ovulation and inflammation are associated with decreased incidence of ovarian cancer development. The purpose of this article is to review the major studies performed in laying hen model of ovarian cancer and discuss how these studies shape our current understanding of the pathophysiology, prevention, and treatment of epithelial ovarian cancer.
Project description:Genetic selection for earlier sexual maturation and extended production cycles in laying hens has significantly improved reproductive efficiency. While limited emphasis has been placed on the underlying physiological changes, we hypothesize that modifications in the control of the hypothalamic-pituitary gonadal (HPG) axis have occurred. Thus, three strains of White leghorn derivatives were followed from hatch to 100 weeks of age (woa), including Lohmann LSL-lite (n = 120) as current commercial hens, heritage Shaver White leghorns (n = 100) as 2000s commercial equivalents, and Smoky Joe hens (n = 68) as 1960s commercial equivalents. Body weight (BW) and egg production were monitored, and blood samples were collected throughout to monitor estradiol (E2) concentrations. Tissue samples were collected at 12, 17, 20, 25, 45, 60, 75, and 100 woa to capture changes in mRNA levels of key genes involved in the HPG axis and monitor ovarian follicular pools. All hens, regardless of strain, age or photoperiod laid their first egg within a 64-gram BW window and, as E2 levels increased prior to photostimulation (PS) in Lohmann and Shaver hens, a metabolic trigger likely induced sexual maturation. However, increased levels of Opsin 5 (OPN5) were observed during the maturation period. Although an elevation in gonadotrophin-releasing hormone I (GnRH-I) mRNA levels was associated with early maturation, no changes in gonadotrophin-inhibitory hormone (GnIH) mRNA levels were observed. Nonetheless, a significant shift in pituitary sensitivity to GnRH was associated with maturation. Throughout the trial, Lohmann, Shaver, and Smoky Joe hens laid 515, 417, and 257 eggs, respectively (p < 0.0001). Results show that the extended laying persistency in Lohmann hens was supported by sustained pituitary sensitivity to GnRH-I, recurrent elevations in follicle-stimulating hormone (FSH) mRNA levels, and five cyclical elevations in E2 levels. This was also associated with a consistently higher pool of small white ovarian follicles. In summary, our results demonstrate first that, regardless of photoperiodic cues, meeting a specific narrow body weight threshold is sufficient to initiate sexual maturation in Leghorn chicken derivatives. Furthermore, recurrent increases in E2 and FSH may be the key to sustain extended laying period, allowing modern layers to double their reproductive capacity compared to their 1960s-counterparts.