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International Journal of Molecular... Aug 2022Vitelline membrane proteins (VMPs) are the main proteins that form the inner shell (vitelline membrane layer) of insect eggs and are an integral part of egg formation...
Vitelline membrane proteins (VMPs) are the main proteins that form the inner shell (vitelline membrane layer) of insect eggs and are an integral part of egg formation and embryo development. Here, we characterized the molecular structure and expression patterns of the gene and analyzed its reproductive functions in diamondback moth, (L.), a worldwide migratory pest of cruciferous plants. The gene was shown to be a single exon gene that contained an open reading frame of 852 base pairs (bp) encoding 283 amino acids. Both qPCR and western blot analyses showed that VMP26 was specifically expressed in female adults and was significantly highly expressed in the ovary. Further anatomical analysis indicated that the expression level of in the ovarian tube with an incomplete yolk was significantly higher than that in the ovarian tube with a complete yolk. CRISPR/Cas9-induced knockout successfully created two homozygous strains with 8- and 46-bp frameshift mutations. The expression deficiency of the VMP26 protein was detected in the mutant strains using immunofluorescence and western blot. No significant difference was found in the number of eggs laid within three days between wild and mutant individuals, but there was a lower egg hatchability. The loss of the gene changed the mean egg size, damaged the structure of the vitelline membrane, and increased the proportion of abnormal eggs due to water loss, resulting in egg collapse. This first analysis of the roles of the gene in the oocyte formation and embryonic development of , using CRISPR/Cas9 technology, provides a basis for screening new genetic control targets of .
Topics: Animals; CRISPR-Cas Systems; Egg Proteins; Female; Moths; Mutagenesis; Vitelline Membrane
PubMed: 36076934
DOI: 10.3390/ijms23179538 -
Child's Nervous System : ChNS :... Jan 2023Proximal ventricular shunt catheter occlusion remains a problematic cause of shunt malfunction, and there is no consistent in vivo or in vitro model to help clinicians...
PURPOSE
Proximal ventricular shunt catheter occlusion remains a problematic cause of shunt malfunction, and there is no consistent in vivo or in vitro model to help clinicians and researchers study this phenomenon.
METHODS
An in vitro model utilizing standard proximal ventricular catheter and biological occluding agents mimicking choroid plexus was designed, constructed, and calibrated to occlude consistently within a specified timeframe. Hydrostatic pressure differential of 100 cmHO was used as a driving force to generate flow through the catheter. Chalaza and vitelline membranes were harvested from avian eggs and used as occluding agents. Successful occlusion was defined as a greater than 90% reduction in volumetric flow rate through distal outlet. Histological sections of occluded catheters were performed and interpreted by a neuropathologist.
RESULTS
Initial trials demonstrated successful standard catheter occlusion within 24 h using chalaza, vitelline membrane, and combination treatments. Repeat trials demonstrated consistency in successful occlusion within 5 min utilizing only vitelline membrane treatment. Histopathology demonstrated the vitelline membrane to consist of a thin, superficial layer of extraembryonic ectoderm; the chalaza was observed to consist of strands of mucin protein.
CONCLUSIONS
An in vitro model of proximal ventricular shunt catheter occlusion was developed and calibrated for successful occlusion within 5 min. Future studies may utilize this model to rapidly test occlusion-resistant shunt designs and de-obstruction techniques.
Topics: Humans; Cerebrospinal Fluid Shunts; Hydrocephalus; Equipment Design; Choroid Plexus; Catheters; Ventriculoperitoneal Shunt
PubMed: 36169702
DOI: 10.1007/s00381-022-05689-z -
The International Journal of... 2018This paper provides a brief account of some aspects of the career of Ruth Bellairs using selected examples from her research publications, with the emphasis being placed...
This paper provides a brief account of some aspects of the career of Ruth Bellairs using selected examples from her research publications, with the emphasis being placed on the early stages of chick embryo development, and in particular, on cell migration. Topics include the role of Hensen's node, the vitelline membrane, the structure and segmentation of somites, the tail bud and the Wolffian duct. Her research approach has involved embryo culture, experimental surgery, transmission and scanning electron microscopy, time-lapse filming and immunostaining techniques.
Topics: Animals; Cell Movement; Chick Embryo; Chickens; Embryo Culture Techniques; Embryology; Embryonic Induction; England; History, 20th Century; History, 21st Century; Mesoderm; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Organizers, Embryonic
PubMed: 29616723
DOI: 10.1387/ijdb.180028rb -
Poultry Science Sep 2017Vitelline membrane (VM) is a multilayered structure that surrounds the egg yolk serving to separate the yolk and the white. Due to its poor solubility in aqueous-based...
Vitelline membrane (VM) is a multilayered structure that surrounds the egg yolk serving to separate the yolk and the white. Due to its poor solubility in aqueous-based media, VM proteins and their biological properties have not been fully defined. In the current study, VM was hydrolyzed using different enzymes under the optimum hydrolysis conditions. Antioxidant and anti-inflammatory properties were evaluated in chemical and cellular models. Flavourzyme- and trypsin-treated samples showed the highest radical scavenging and ferric ion reducing effect (31% and 20 μM of Trolox equivalents/mg, respectively). In cellular studies, all VM hydrolysates were cyto-compatible and inhibited nitric oxide production by RAW264.7 macrophage cells significantly. Lipopolysaccharide-stimulated up-regulation of pro-inflammatory cytokines in RAW264.7 cells was suppressed by flavourzyme-treated VM. These results revealed that enzymatic hydrolysis of VM is a promising approach to produce peptides with several bioactivities (free radical scavenging, metal chelation, and anti-inflammatory) as valuable ingredients for cosmeceuticals and nutraceuticals.
Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Chickens; Egg Proteins; Hydrolysis; Models, Chemical; Protein Hydrolysates; Vitelline Membrane
PubMed: 28854750
DOI: 10.3382/ps/pex125 -
Seminars in Cell & Developmental Biology Feb 2017Twenty-seven serpins belonging to clade A, B, C, D, E, F, G, H and I serpins are currently referenced in chicken genome databases. Phylogenetic analysis of chicken... (Review)
Review
Twenty-seven serpins belonging to clade A, B, C, D, E, F, G, H and I serpins are currently referenced in chicken genome databases. Phylogenetic analysis of chicken serpins revealed that ovalbumin (Serpinb14) and its paralogs ovalbumin-related protein Y (Serpinb14b) and ovalbumin-related protein X (Serpinb14c) are found in bird species. These clade B serpins are specifically expressed in reproductive tissues and exported in the egg where they constitute major protein components. These data suggest that these three paralogs have probably appeared in birds to face new environments and ensure the extra-uterine development of an embryo in a shell egg. Twelve other serpins have been identified in the newly produced egg, some of them having a specific distribution in the respective egg structures (eggshell, egg white, vitelline membrane and egg yolk). The physiological role of these egg serpins remain largely unexplored, but there is increasing evidence in literature or by homologies with their mammalian counterparts, that some of them participate in cell proliferation, tissue remodeling and/or angiogenesis associated with folliculogenesis and development of extraembryonic structures, eggshell biomineralization, egg defense and nutrition of the embryo. A better knowledge of the phylogenetic evolution of these 15 serpins in other oviparous species, on their egg distribution, on their regulation during embryonic development (activation/degradation/transfer) and on their functional specificity, is needed to better appreciate their role and their bird-specificity. These review shed light on the multiple possibilities that offer the avian egg model to study the role of serpins in reproduction and developmental biology.
Topics: Animals; Chickens; Evolution, Molecular; Models, Molecular; Ovum; Phylogeny; Serpins
PubMed: 27565683
DOI: 10.1016/j.semcdb.2016.08.019 -
Scientific Reports Nov 2020In this study, we aimed to perform structural and proteomic analysis of the vitelline membrane (VM) of two species birds belonging to the family Turdidae: blackbird...
In this study, we aimed to perform structural and proteomic analysis of the vitelline membrane (VM) of two species birds belonging to the family Turdidae: blackbird (Turdus merula) and song thrush (Turdus philomelos). We performed structural analyses using scanning electron microscopy. The VM proteins were identified and compared to the best-known chicken VM proteins. According to our results, VM of both species has a typical three-layered structure: the outer layer, inner layer, and the continuous membrane between them. An unusual observation was the finding of "convexity" formed by the inner layer in blackbird. The role of these convex structures is not known, but they can be typical for the species and can be used in their identification. In addition, we identified two proteins in the VM of both species of birds, of which U3KEZ1 FICAL was not previously identified in any other bird species, and the U3JXV8 FICAL protein was confirmed only once in cockatiel parrot VM. The function of these proteins is not exactly known, but their structure shows similarities to the SERPIN proteins that are involved in microbiological defense, i.e., they are immune proteins. This study contributes to the current knowledge about the structure and composition of proteins of VM, especially because similar analyses have never been performed for Turdidae family. Knowledge of the structure and specific proteins of blackbird and song thrush VM can be beneficial in research on ecology and bird biology and also helpful in developing noninvasive and nongenetic identification methods.
Topics: Animals; Chickens; Egg Proteins; Egg Yolk; Female; Immune System; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Proteome; Songbirds; Species Specificity; Vitelline Membrane
PubMed: 33168893
DOI: 10.1038/s41598-020-76559-4 -
Bioengineering (Basel, Switzerland) May 2023The lipid layer surrounding the vitelline membrane of insect eggs has a critical role in the waterproofing and desiccation resistance of embryos. However, this lipid...
The lipid layer surrounding the vitelline membrane of insect eggs has a critical role in the waterproofing and desiccation resistance of embryos. However, this lipid layer also prevents the flux of chemicals into the embryos, such as cryoprotectants, which are required for successful cryopreservation. The permeabilization studies of silkworm embryos remain insufficient. Therefore, in this study, we developed a permeabilization method to remove the lipid layer in the silkworm, , and examined factors affecting the viability of dechorionated embryos, including the types and exposure times of chemicals and embryonic stages. Among the chemicals used, hexane and heptane were effective for permeabilization, whereas Triton X-100 and Tween-80 were less effective. Regarding the embryonic stages, there were significant differences between 160 and 166 h after egg laying (AEL) at 25 °C. Consequently, we found that the treatment of 160 AEL embryos with hexane for 30 s was the best condition for the permeability and viability of embryos, in which over 62% of the permeabilized embryos grew up to the second larval instar and their moths could lay fertilized eggs. Our method can be used for various purposes, including permeability investigations using other chemicals and embryonic cryopreservation.
PubMed: 37237633
DOI: 10.3390/bioengineering10050563 -
Wiley Interdisciplinary Reviews.... 2014The pathway that generates the dorsal-ventral (DV) axis of the Drosophila embryo has been the subject of intense investigation over the previous three decades. The... (Review)
Review
UNLABELLED
The pathway that generates the dorsal-ventral (DV) axis of the Drosophila embryo has been the subject of intense investigation over the previous three decades. The initial asymmetric signal originates during oogenesis by the movement of the oocyte nucleus to an anterior corner of the oocyte, which establishes DV polarity within the follicle through signaling between Gurken, the Drosophila Transforming Growth Factor (TGF)-α homologue secreted from the oocyte, and the Drosophila Epidermal Growth Factor Receptor (EGFR) that is expressed by the follicular epithelium cells that envelop the oocyte. Follicle cells that are not exposed to Gurken follow a ventral fate and express Pipe, a sulfotransferase that enzymatically modifies components of the inner vitelline membrane layer of the eggshell, thereby transferring DV spatial information from the follicle to the egg. These ventrally sulfated eggshell proteins comprise a localized cue that directs the ventrally restricted formation of the active Spätzle ligand within the perivitelline space between the eggshell and the embryonic membrane. Spätzle activates Toll, a transmembrane receptor in the embryonic membrane. Transmission of the Toll signal into the embryo leads to the formation of a ventral-to-dorsal gradient of the transcription factor Dorsal within the nuclei of the syncytial blastoderm stage embryo. Dorsal controls the spatially specific expression of a large constellation of zygotic target genes, the Dorsal gene regulatory network, along the embryonic DV circumference. This article reviews classic studies and integrates them with the details of more recent work that has advanced our understanding of the complex pathway that establishes Drosophila embryo DV polarity. For further resources related to this article, please visit the WIREs website.
CONFLICT OF INTEREST
The authors have declared no conflicts of interest for this article.
Topics: Animals; Body Patterning; Drosophila; Epistasis, Genetic; Gene Expression Regulation, Developmental; Gene Regulatory Networks
PubMed: 25124754
DOI: 10.1002/wdev.138 -
BioRxiv : the Preprint Server For... May 2023The eggshell of the fruit fly is a useful model for understanding the synthesis of a complex extracellular matrix. The eggshell is synthesized during mid-to-late...
The eggshell of the fruit fly is a useful model for understanding the synthesis of a complex extracellular matrix. The eggshell is synthesized during mid-to-late oogenesis by the somatic follicle cells that surround the developing oocyte. We previously reported that female flies mutant for the gene ( ) are sterile, but the underlying cause of the sterility remained unknown. In this study, we examined the role of in eggshell synthesis. We show that eggs laid by mutant females are fertilized but arrest early in embryogenesis, and that the innermost layer of the eggshell, the vitelline membrane, is abnormally permeable to dye in these eggs. In addition, the major vitelline membrane proteins fail to become crosslinked by nonreducible bonds, a process that normally occurs during egg activation following ovulation, as evidenced by their solubility and detection by Western blot in laid eggs. In contrast, the Cp36 protein, which is found in the outer chorion layers of the eggshell, becomes crosslinked normally. To link the expression pattern with these phenotypes, we show that is expressed in the ovarian follicle cells beginning in mid-oogenesis, and, importantly, that all mutant eggshell phenotypes could be recapitulated by selective knockdown of expression in the follicle cells. To determine whether expression was required for the crosslinking itself, we performed in vitro activation and crosslinking experiments. The vitelline membranes of control egg chambers could become crosslinked either by incubation in hyperosmotic medium, which activates the egg chambers, or by exogenous peroxidase and hydrogen peroxide. In contrast, neither treatment resulted in the crosslinking of the vitelline membrane in mutant egg chambers. These results indicate that expression in the follicle cells is necessary for vitelline membrane proteins to serve as substrates for peroxidase-mediated cross-linking at the end of oogenesis.
PubMed: 37163052
DOI: 10.1101/2023.04.25.538335