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Developmental Biology May 2019Stereotyped left-right asymmetry both in external and internal organization is found in various animals. Left-right symmetry is broken by the neurula rotation in the...
Stereotyped left-right asymmetry both in external and internal organization is found in various animals. Left-right symmetry is broken by the neurula rotation in the ascidian, Halocynthia roretzi. Neurula embryos rotate along the anterior-posterior axis in a counterclockwise direction, and the rotation stops when the left side of the embryo is oriented downwards, resulting in contact of the left-side epidermis with the vitelline membrane at the bottom of perivitelline space. Then, such contact induces the expression of nodal and its downstream Pitx2 gene in the left-side epidermis. Vitelline membrane is required for the promotion of nodal expression. Here, we showed that a chemical signal from the vitelline membrane promotes nodal gene expression, but mechanical stimulus at the point of contact is unnecessary since the treatment of devitellinated neurulae with an extract of the vitelline membrane promoted nodal expression on both sides. The signal molecules are already present in the vitelline membranes of unfertilized eggs. These signal molecules are proteins but not sugars. Specific fractions in gel filtration chromatography had the nodal promoting activity. By mass spectrometry, we selected 48 candidate proteins. Proteins that contain both a zona pellucida (ZP) domain and epidermal growth factor (EGF) repeats were enriched in the candidates of the nodal inducing molecules. Six of the ZP proteins had multiple EGF repeats that are only found in ascidian ZP proteins. These were considered to be the most viable candidates of the nodal-inducing molecules. Signal molecules are anchored to the entire vitelline membrane, and contact sites of signal-receiving cells are spatially and mechanically controlled by the neurula rotation. In this context, ascidians are unusual with respect to mechanisms for specification of the left-right axis. By suppressing formation of epidermis monocilia, we also showed that epidermal cilia drive the neurula rotation but are dispensable for sensing the signal from the vitelline membrane.
Topics: Animals; Body Patterning; Cell Extracts; Cilia; Egg Proteins; Embryo, Nonmammalian; Epidermis; Gene Expression Regulation, Developmental; Glycosylation; Nodal Protein; Protein Domains; Quinazolinones; Rotation; Signal Transduction; Sugars; Urochordata; Vitelline Membrane
PubMed: 30710513
DOI: 10.1016/j.ydbio.2019.01.016 -
Parasites & Vectors Aug 2023Stephanofilaria stilesi is a vector-borne filarioid nematode of cattle in North America that is transmitted via the hematophagous horn fly (Haematobia irritans)...
BACKGROUND
Stephanofilaria stilesi is a vector-borne filarioid nematode of cattle in North America that is transmitted via the hematophagous horn fly (Haematobia irritans) intermediate host. Despite being relatively common, little attention has been given to a thorough description of S. stilesi lesions and the potential integration of pathological and molecular diagnostic findings to confirm infection.
METHODS
To characterize the cutaneous lesions caused by S. stilesi in cattle (Bos taurus taurus and Bos taurus indicus), skin of the ventral abdominal midline was collected from 22 animals during postmortem examination. Skin samples were processed for histology, transmission electron microscopy (TEM), DNA extraction, PCR, and Sanger sequencing targeting molecular markers cytochrome oxidase c subunit 1 (cox1), 12S, 18S rDNA, and 28S rDNA.
RESULTS
Macroscopically, lesions ranged from 5 × 4 cm to 36 × 10 cm, consisting of one large single lesion, or two to four ovoid areas at the ventral abdominal midline, surrounding the umbilicus. Each lesion presented as ulcerative dermatitis with dry, serocellular crusts, or alopecic and lichenified areas. Histologically, eosinophilic, neutrophilic, and ulcerative dermatitis with furunculosis, folliculitis, and epidermal hyperplasia was observed. Cross sections of adult nematodes were identified in ~ 60% of the cases (n = 13) within intact follicles, sebaceous ducts, crusts, and areas of furunculosis. Stephanofilaria first-stage larvae (L1) were observed in five cases within "vitelline membranes" in the superficial dermis and crusts. Ultrastructurally, the L1 cross sections were compounded of smooth multilayered cuticle and somatic cells. The "vitelline membrane" is a tri-layered membrane where L1 are suspended in a matrix. Stephanofilaria stilesi DNA was found in 5 out of the 13 cases in which adults or L1 were histologically observed (38%) and in 1 out of the 9 cases without adults or L1 present (11%). Phylogenetic analyses suggest a closer relationship of the genus Stephanofilaria with Thelazioidea, instead of the family Filariidae (Filarioidea), in which it has been historically allocated.
CONCLUSIONS
Our study improved the characterization of lesions and described ultrastructural findings of S. stilesi and highlights that molecular tools should be utilized in combination with histology for improved diagnostic resolution.
Topics: Animals; Cattle; Phylogeny; Furunculosis; Filarioidea; Dermatitis; Muscidae; DNA, Ribosomal
PubMed: 37573424
DOI: 10.1186/s13071-023-05905-y -
Animals : An Open Access Journal From... Mar 2021The yolk is the principal part of the egg that contains vitamins, minerals, lipids, and proteins which are essential for embryo development and hatching. The egg yolk...
The yolk is the principal part of the egg that contains vitamins, minerals, lipids, and proteins which are essential for embryo development and hatching. The egg yolk contains significant amounts of lipoproteins, triacylglycerides, and cholesterol, whose dynamics are indistinct during embryogenesis. The effects of cholesterol on the yolk protein abundance, intensity, and function are ill-defined during embryonic development. Using two-dimensional gel electrophoresis, eggs with respective high and low cholesterol protein abundance were investigated after 0, 2, 6, and 13 days of embryogenesis and further analyzed by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. The results revealed that the vitellogenin proteins are the most abundant egg yolk protein that showed proximity and a high degree of variation in isoelectric point and molecular weight. The results demonstrated increased expression of vitellogenin-1 and vitellogenin-3 at two days and vitellogenin-2 protein at 13 days of embryogenesis in both egg types. The ovoinhibitor, immunoglobulin lambda light chain precursor, Ig-gamma (clone-36 chicken), and beta-2-glycoprotein-1 precursor proteins were significantly expressed in high cholesterol eggs while haptoglobin protein PIT-54 and vitelline membrane outer layer proteins intensities were significant in low cholesterol eggs at two days of embryogenesis. The high cholesterol eggs showed a modest increase in egg weight, yolk weight, albumen height, yolk color, and egg strength relative to the low cholesterol eggs. The gene ontology enrichment analysis revealed that the differentially expressed proteins such as vitellogenin proteins were involved in lipid transport and lipid localization biological processes and showed nutrient reservoir activity function. The ovotransferrin regulated the biological processes of plasminogen activation and extracellular matrix disassembly and characterized the anchored component of the plasma membrane. The ovoinhibitor protein was involved in response to mineralocorticoid and corticosterone biological processes whereas the vitellin membrane outer layer protein constituted the extracellular exosome, extracellular organelle, and membrane-bounded vesicle cellular components. Collectively, our study revealed yolk protein abundance, molecular function, cellular components, and biological processes and concluded that yolk protein intensities were significantly altered by cholesterol concentration.
PubMed: 33803097
DOI: 10.3390/ani11030744 -
Poultry Science Dec 2023The study aimed to analyze the hatching egg and physiochemical features of eggshells, thick albumen, amniotic fluid, and yolk during the incubation of Ross 308 chicken...
The study aimed to analyze the hatching egg and physiochemical features of eggshells, thick albumen, amniotic fluid, and yolk during the incubation of Ross 308 chicken eggs. Eggs (n = 755) were incubated for 21 d. Quality analysis of fresh eggs was performed. Eggshells, albumen, and yolk were collected from fresh eggs and incubation d 1, 7, and 14. Eggshell thickness and strength, pH, vitelline membrane strength, fatty acid (FA) in the yolk, pH, viscosity, lysozyme activity, and crude protein content in thick albumen and amniotic fluid were analyzed. Hatching parameters were calculated. Egg weight loss was constant (8.04% overall). Lower egg surface temperature was found on d 7 compared to d 4, 14, and 18. A lower thickness of posthatch eggshells was found. The strength of the vitelline membrane significantly decreased within 24 h (by over 58%). During incubation, there was a decrease in thick albumen/amniotic fluid pH; an opposite trend was found in yolk pH. The vitelline membrane strength was negatively correlated with the albumen pH. Lysozyme activity was higher in fresh thick albumen and up to 2 wk of incubation. On d 7, the lowest activity was found in the amniotic fluid. On d 14, lysozyme activity increased in amniotic fluid. The higher viscosity of the thick albumen was demonstrated on d 7 and 14 of incubation. The lowest viscosity in amniotic fluid was found on the same days. Crude protein content was higher in thick albumen (d 7 and 14) and lowest in amniotic fluid on d 7. The FA content changed between d 0 and 14. The results indicate different use of FA, where PUFA decreased. Eggshell is used in the last week of incubation. The thick albumen is reduced, while the biological value of amniotic fluid is increasing. Lysozyme activity, viscosity, and crude protein content may be interdependent. It may indicate the flow of substances and the transfer of functions from the thick albumen to the amniotic fluid during chicken embryogenesis.
Topics: Animals; Chickens; Egg Shell; Muramidase; Amniotic Fluid; Ovum; Albumins; Fatty Acids; Embryonic Development; Egg Yolk; Eggs
PubMed: 37832191
DOI: 10.1016/j.psj.2023.103119 -
Changes in physicochemical parameters of duck eggs and extra-embryonic structures during incubation.Animal : An International Journal of... Dec 2023Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the...
Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the embryo to hazards. The study aimed to analyse the physicochemical properties of eggshell, yolk, thick albumen (TA), and amniotic fluid (AF) of incubated hatching eggs from 52-week-old Cherry Valley ducks. The morphological features of 18 fresh eggs were analysed. Over 28 days, a total of 800 eggs underwent incubation. Eggshell surface temperature and egg weight loss were measured on days 1, 4, 7, 10, 14, 18, 21, and 25. Eggshell, TA, AF, and yolk were collected from eggs at incubation days 1-21 (every week). TA was collected on days 0, 1, and 7, while AF on days 7, 14, and 21. The analysis covered a range of physicochemical parameters. Eggshell thickness decreased with incubation, reaching its lowest point posthatch (P < 0.001). The highest pH for TA was recorded on day 1, while the lowest was on day 7 when comparing days 0, 1, and 7 (P < 0.001). TA pH was consistently higher than in AF (P < 0.001). However, the pH of TA was the highest on day 1 and the lowest on day 7 (P < 0.001). Yolk pH increased from days 1 to 21 (P < 0.001). There was also a noticeable in egg weight loss (0.34% daily) (P < 0.001). Vitelline membrane strength decreased from day 0 to day 1 (P < 0.001). Lysozyme activity in thick albumen on day 7 was higher than on days 0 and 1 (P < 0.001). Lysozyme activity in AF was higher on day 21 than days 7 and 14 (P < 0.001). TA viscosity was highest on day 0 and lowest on day 1, compared to other days (P < 0.001). AF viscosity and CP content exhibited an increase on day 21 as compared to days 7 and 14 (P < 0.001). The CP content in TA was notably higher on day 7 than on days 0 and 1 (P < 0.001). Polyunsaturated fatty acids declined, while monounsaturated and transfatty acids increased (P < 0.001). Viscosity and lysozyme activity increased on day 7 in TA and day 21 in AF. TA and the amniotic cavity appeared to facilitate the transfer of substances, particularly CP. Viscosity could be an indicator for optimising incubation conditions, as incorrect changes can affect embryo mortality. The results showed the different utilisation of nutrients, such as fatty acids. It could support research on the in-ovo administration of various substances.
Topics: Animals; Ducks; Muramidase; Ovum; Egg Shell; Weight Loss; Eggs; Chickens
PubMed: 37981451
DOI: 10.1016/j.animal.2023.101024 -
FEBS Open Bio Mar 2023C-mannosylation is a rare type of protein glycosylation whereby a single mannose is added to the first tryptophan in the consensus sequence Trp-Xaa-Xaa-Trp/Cys (in which...
C-mannosylation is a rare type of protein glycosylation whereby a single mannose is added to the first tryptophan in the consensus sequence Trp-Xaa-Xaa-Trp/Cys (in which Xaa represents any amino acid). Its consensus sequence is mainly found in proteins containing a thrombospondin type-1 repeat (TSR1) domain and in type I cytokine receptors. In these proteins, C-mannosylation affects protein secretion, intracellular localization, and protein stability; however, the role of C-mannosylation in proteins that are not type I cytokine receptors and/or do not contain a TSR1 domain is less well explored. In this study, we focused on human vitelline membrane outer layer protein 1 homolog (VMO1). VMO1, which possesses two putative C-mannosylation sites, is a 21-kDa secreted protein that does not contain a TSR1 domain and is not a type I cytokine receptor. Mass spectrometry analyses revealed that VMO1 is C-mannosylated at Trp but not at Trp . Although C-mannosylation does not affect the extracellular secretion of VMO1, it destabilizes the intracellular VMO1. In addition, a structural comparison between VMO1 and C-mannosylated VMO1 showed that the modification of the mannose changes the conformation of three loops in VMO1. Taken together, our results demonstrate the first example of C-mannosylation for protein destabilization of VMO1.
Topics: Humans; Glycosylation; Mannose; Vitelline Membrane; Protein Transport; Receptors, Cytokine
PubMed: 36680395
DOI: 10.1002/2211-5463.13561 -
PloS One 2020Of all the known oviparous taxa, female birds lay the most diverse types of eggs that differ in terms of shape, shell pigmentation, and shell structure. The pigmentation... (Comparative Study)
Comparative Study
Characterization of structure and protein of vitelline membranes of precocial (ring-necked pheasant, gray partridge) and superaltricial (cockatiel parrot, domestic pigeon) birds.
Of all the known oviparous taxa, female birds lay the most diverse types of eggs that differ in terms of shape, shell pigmentation, and shell structure. The pigmentation of the shell, the weight of the egg, and the composition of the yolk correlate with environmental conditions and the needs of the developing embryos. In this study, we analyzed the structure and protein composition of the vitelline membrane (VM) of ring-necked pheasant, gray partridge, cockatiel parrot, and domestic pigeon eggs. We found that the VM structure is characteristic of each species and varies depending on whether the species is precocial (ring-necked pheasant and gray partridge) or superaltrical (cockatiel parrot and domestic pigeon). We hypothesize that a multilayer structure of VM is necessary to counteract the aging process of the egg. The multilayer structure of VM is only found in species with a large number of eggs in one clutch and is characterized by a long incubation period. An interesting discovery of this study is the three-layered VM of pheasant and partridge eggs. This shows that the formation of individual layers of VM in specific sections of the hen's reproductive system is not confirmed in other species. The number of protein fractions varied between 19 and 23, with a molecular weight ranging from 15 to 250 kDa, depending on the species. The number of proteins identified in the VM of the study birds' eggs is as follows: chicken-14, ring-necked pheasant-7, gray partridge-10, cockatiel parrot-6, and domestic pigeon-23. The highest number of species-specific proteins (21) was detected in the VM of domestic pigeon. This study is the first to present the structure and protein composition in the VM of ring-necked pheasant, gray partridge, cockatiel parrot, and domestic pigeon eggs. In addition, we analyzed the relationship between the hatching specification of birds and the structure of the VM.
Topics: Animals; Cockatoos; Columbidae; Egg Proteins; Female; Galliformes; Male; Microscopy, Electron, Scanning; Molecular Weight; Protein Interaction Maps; Proteomics; Species Specificity; Vitelline Membrane
PubMed: 31999757
DOI: 10.1371/journal.pone.0228310 -
Lipids Jun 2016The vitelline membrane (VM) encloses the chicken egg yolk, separating it from albumen. The VM weakens during storage, and dietary lipid modification significantly...
The vitelline membrane (VM) encloses the chicken egg yolk, separating it from albumen. The VM weakens during storage, and dietary lipid modification significantly affects its strength. However, no studies have characterize the fatty acyl residue (FA) composition of the VM, and reports of VM isolation and quantified lipid content are inconsistent. Therefore, the objectives of this study were: (1) to develop a washing and isolation method that removes residual yolk from VM without damage; (2) to determine the FA and lipid composition of CLA-rich egg yolk VM, relative to controls; (3) to determine the effect of 20 days of refrigeration on VM FA and lipid composition. To determine VM FA and lipid composition, 36 hens received either a corn-soybean meal-based control diet ("Control"), or the Control supplemented with either 10 % soy oil ("Soy control"), or 10 % CLA-rich soy oil ("CLA") for 30 days. VM were analyzed the day of collection ("fresh"), or after 20 days of refrigeration ("refrigerated"). There were no differences in FA compositions of fresh and refrigerated membranes within a treatment. CLA-rich yolk VM contains CLA, greater SFA, and significantly greater DHA relative to controls. Direct MALDI-TOF-MS identified 15 phosphatidylcholines, three phosphatidylethanolamines, one sphingomyelin, and 15 triacylglycerols in VM. Lipid species that showed significant differences among egg types included nine phosphatidylcholines and six triacylglycerols. MALDI analysis indicated significant differences in nine lipid classes on the VM inner layer. After refrigeration, five lipid classes on the inner layer and seven lipid classes on the outer layer had statistically significant differences among VM types.
Topics: Animal Feed; Animals; Chickens; Egg Yolk; Linoleic Acid; Membrane Lipids; Refrigeration; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Vitelline Membrane
PubMed: 27108035
DOI: 10.1007/s11745-016-4153-x -
Journal of Innate Immunity 2019The integrated innate immune features of the calcareous egg and its contents are a critical underpinning of the remarkable evolutionary success of the Aves clade.... (Review)
Review
Dynamics of Structural Barriers and Innate Immune Components during Incubation of the Avian Egg: Critical Interplay between Autonomous Embryonic Development and Maternal Anticipation.
The integrated innate immune features of the calcareous egg and its contents are a critical underpinning of the remarkable evolutionary success of the Aves clade. Beginning at the time of laying, the initial protective structures of the egg, i.e., the biomineralized eggshell, egg-white antimicrobial peptides, and vitelline membrane, are rapidly and dramatically altered during embryonic development. The embryo-generated extra-embryonic tissues (chorioallantoic/amniotic membranes, yolk sac, and associated chambers) are all critical to counteract degradation of primary egg defenses during development. With a focus on the chick embryo (Gallus gallus domesticus), this review describes the progressive transformation of egg innate immunity by embryo-generated structures and mechanisms over the 21-day course of egg incubation, and also discusses the critical interplay between autonomous development and maternal anticipation.
Topics: Animals; Antimicrobial Cationic Peptides; Chick Embryo; Chickens; Egg Shell; Embryonic Development; Female; Immunity, Innate; Maternal-Fetal Exchange; Ovum; Pregnancy; Vitelline Membrane
PubMed: 30391943
DOI: 10.1159/000493719 -
Development Genes and Evolution Sep 2017The appendicularian Oikopleura dioica is a planktonic chordate that retains a tadpole shape throughout its life. Its simple and transparent body, invariant cell...
The appendicularian Oikopleura dioica is a planktonic chordate that retains a tadpole shape throughout its life. Its simple and transparent body, invariant cell lineages, fast development and available genome and transcriptome resources make it a promising model organism for research in developmental biology. However, large-scale analysis of gene expression in O. dioica is limited owing to the laborious and time-consuming process of manual removal of the vitelline membrane, because devitellinisation of pre-hatching embryos causes failure of normal development. Therefore, in this study, modified procedures were developed for whole-mount in situ hybridisation (WISH) and immunohistochemistry (WIHC). This protocol enables rapid mRNA or protein detection without a manual devitellination step for each specimen. The critical procedure is brief treatment of the vitelline membrane of living embryos with 0.05% actinase E before fixation. Two minutes of treatment was optimal for the penetration of antisense RNA probes and antibodies through the vitelline membrane. This WISH protocol was applicable for chromogenic and fluorescent tyramide signal amplification reactions. Using the new protocol, we found eight genes with tissue-specific expression in the tail muscle, trunk epidermis, heart, pharynx, oesophagus, stomach or gill openings of developing larvae. This procedure also allowed for the detection of exogenous FLAG-tagged histone-enhanced green fluorescent protein by WIHC using anti-FLAG antibody. This study provides a useful and convenient tool for studying spatial and temporal gene expression patterns in this simple chordate model and should facilitate handling large amounts of genetic data from transcriptome-based approaches and other techniques such as treatments with chemical inhibitors.
Topics: Animals; Cell Lineage; Embryo, Nonmammalian; Gene Expression Regulation, Developmental; Green Fluorescent Proteins; Immunohistochemistry; In Situ Hybridization; Organ Specificity; Urochordata; Vitelline Membrane
PubMed: 28752326
DOI: 10.1007/s00427-017-0588-1