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Applied Microscopy Jul 2022We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also...
We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also known as Norman's lampeye using light and electron microscopes. The fertilized eggs with narrow perivitelline space were found to be spherical and demersal, additionally containing small oil droplets in the vitelline membrane. Further, a bundle of adhesive filaments was observed to be present on one side of the fertilized egg. These filaments possessed remarkably high elasticity and were approximately 1-3 mm in length. The size of the fertilized egg was determined to be about 1.49 ± 0.07 mm (n = 30). The outer surface appeared smooth, and adhesive filaments originating at different location of the surface of the envelope were found to be distributed around the egg envelope and were joined together to form a single long bundle in scanning electron microscopic observation. A peak-like structure formed of several straight wrinkles was observed around the micropyle. However, the complete structure of the micropyle could not be studied due to the depth at which it was located. Additionally, the total thickness of the egg envelope was ascertained to be approximately12.5-14.5 μm. The egg envelope consisted of two distinct layers, an outer electron dense layer and an inner lamellar layer, further consisting of 10 sublayers of varying thicknesses. Collectively, it was observed that the morphological characteristics of the fertilized egg, fine structures surrounding the micropyle, outer surface, adhesive structure consisting adhesive filaments, and sections of fertilized egg envelope displayed species specificity.
PubMed: 35831688
DOI: 10.1186/s42649-022-00075-0 -
Brazilian Journal of Biology = Revista... Feb 2016The objective of this study was to describe the embryonic and larval development of Brycon amazonicus, featuring the main events up to 50 hours after fertilization (AF)....
The objective of this study was to describe the embryonic and larval development of Brycon amazonicus, featuring the main events up to 50 hours after fertilization (AF). The material was provided by the Aquaculture Training, Technology and Production Center, Presidente Figueiredo (AM). The characterization was based on stereomicroscopic examination of the morphology of eggs, embryos and larvae and comparison with the literature. Matrinxã eggs are free, transparent, and spherical, with a perivitelline space of 0.56 ± 0.3 mm. The successive divisions give rise to cells with 64 blastomeres during the first hour AF. The gastrula stage, beginning 02 h 40 min AF, was characterized by progressive regression cells and the formation of the embryonic axis, leading to differentiation of the head and tail 05 h 30 min AF. From 06 to 09 h AF the somites, notochord, otic and optic vesicles and otoliths were observed, in addition to heart rate and the release of the tail. The larvae hatched at 10 h 30 min AF (29.9 °C), with a total length of 3.56 ± 0.46 mm. Between 19 and 30 h AF, we observed 1) pigmentation and gut formation, 2) branchial arches, 3) pectoral fins, 4) a mouth opening and 5) teeth. Cannibalism was initiated earlier (34 h AF) which was associated with rapid yolk absorption (more than 90% until 50 h AF), signaling the need for an exogenous nutritional source. The environmental conditions (especially temperature) influenced the time course of some events throughout the embryonic and larval development, suggesting the need for further studies on this subject.
Topics: Animals; Characidae; Embryo, Nonmammalian; Embryonic Development
PubMed: 26909629
DOI: 10.1590/1519-6984.13914 -
Animals : An Open Access Journal From... Apr 2021This study aimed to examine the effects of treatment with glucuronic acid (GA) and N-acetyl-D-glucosamine (AG), which are components of hyaluronic acid (HA), during...
Effect of D-Glucuronic Acid and N-acetyl-D-Glucosamine Treatment during In Vitro Maturation on Embryonic Development after Parthenogenesis and Somatic Cell Nuclear Transfer in Pigs.
This study aimed to examine the effects of treatment with glucuronic acid (GA) and N-acetyl-D-glucosamine (AG), which are components of hyaluronic acid (HA), during porcine oocyte in vitro maturation (IVM). We measured the diameter of the oocyte, the thickness of the perivitelline space (PVS), the reactive oxygen species (ROS) level, and the expression of cumulus cell expansion and ROS-related genes and examined the cortical granule (CG) reaction of oocytes. The addition of 0.05 mM GA and 0.05 mM AG during the first 22 h of oocyte IVM significantly increased oocyte diameter and PVS size compared with the control (non-treatment). The addition of GA and AG reduced the intra-oocyte ROS content and improved the CG of the oocyte. GA and AG treatment increased the expression of and in cumulus cells and and in oocytes. In both the chemically defined and the complex medium (Medium-199 + porcine follicular fluid), oocytes derived from the GA and AG treatments presented significantly higher blastocyst rates than the control after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). In conclusion, the addition of GA and AG during IVM in pig oocytes has beneficial effects on oocyte IVM and early embryonic development after PA and SCNT.
PubMed: 33917537
DOI: 10.3390/ani11041034 -
Antibiotics (Basel, Switzerland) Dec 2023(1) Background: Microinjection of zebrafish () embryos offers a promising model for studying the virulence and potential environmental risks associated with . (2)...
(1) Background: Microinjection of zebrafish () embryos offers a promising model for studying the virulence and potential environmental risks associated with . (2) Methods: This work aimed to develop a infection model using two parallel exposition pathways on zebrafish larvae with microinjection into the yolk and the perivitelline space to simultaneously detect the invasive and cytotoxic features of the examined strains. The microinjection infection model was validated with 15 environmental and clinical strains of of various origins, antibiotic resistance profiles, genotypes and phenotypes: both exposition pathways were optimized with a series of bacterial dilutions, different drop sizes (injection volumes) and incubation periods. Besides mortality, sublethal symptoms of the treated embryos were detected and analyzed. (3) Results: According to the statistical evaluation of our results, the optimal parameters (dilution, drop size and incubation period) were determined. (4) Conclusions: The tested zebrafish embryo microinjection infection model is now ready for use to determine the in vivo virulence and ecological risk of environmental .
PubMed: 38136774
DOI: 10.3390/antibiotics12121740 -
International Journal of Reproductive... Mar 2023Phenotypic dysmorphism is not rare to be found in the human oocyte, especially in the perivitelline space, which are among the most important aberration of the extra...
BACKGROUND
Phenotypic dysmorphism is not rare to be found in the human oocyte, especially in the perivitelline space, which are among the most important aberration of the extra cytoplasmic component.
CASE PRESENTATION
The case is of a 30-yr-old woman with no previous pregnancy, attempting an in vitro fertilization treatment for the first time. Given the extraordinary quantity of granular particles found in the perivitelline space, visible after the stripping procedure, it was not possible to establish the presence and position of the first polar body to appreciate the correct oocyte maturation (metaphase 2). Nevertheless, all the eggs were injected by the intracytoplasmic sperm injection. A time lapse incubator was used to perform the entire culture. Hence, a record of 6 days culture video was obtained. Only 2 eggs could fertilize correctly and reach the blastocyst stage on day 6. The embryos were frozen and subsequently transferred as frozen embryo transfer following the next menstrual cycle.
CONCLUSION
The exceptional presence of granular particles in the perivitelline space, which reminds us for aspects and behavior of the granulosa cells, seems to affect the fertilization but not the blastocysts quality. As a matter of fact, the woman, after the embryo transfer, achieved a successful twin live birth.
PubMed: 37122891
DOI: 10.18502/ijrm.v21i3.13202 -
Biomolecules Jan 2023Alveolin is a cortical alveolus proteinase that is secreted in the perivitelline space (PVS) at fertilization to act on the chorion. Purified alveolin is known to induce...
Alveolin is a cortical alveolus proteinase that is secreted in the perivitelline space (PVS) at fertilization to act on the chorion. Purified alveolin is known to induce chorion hardening in vitro by processing zona pellucida B (ZPB), a major chorion component. However, in vivo function of alveolin remains unclear; thus, in this study, the effects of efficiency () at the organism level were investigated using the medaka, . The fertilized eggs were mechanically fragile; however, they developed normally and left offspring as long as they were carefully handled before hatching. A mechanical press test showed that the fertilized eggs were six times more fragile than the wild-type eggs. They were 35% larger owing to the enlarged PVS, 34% thinner, and permeable to even 10 kDa FITC-dextran. These results are consistent with the transmission electron microscopy observation that the periphery of the inner layers was highly porous in the chorion. In chorion hardening, the alveolin-mediated processing of ZPB and the transglutaminase (TGase)-mediated crosslinking of chorion components are the key steps. This study was the first to show that alveolin also processed TGase concomitantly with ZPB, which greatly facilitated the crosslinking. Thus, alveolin was concluded to be the primary trigger for chorion hardening in vivo. Furthermore, fertilization in a balanced salt solution could partially improve the impaired chorion hardening of the eggs fertilized in water, probably through an alveolin-independent mechanism.
Topics: Animals; Oryzias; Peptide Hydrolases; Fertilization; Chorion
PubMed: 36671531
DOI: 10.3390/biom13010146 -
International Journal of Fertility &... Oct 2020Testicular cell conditioned medium (TCCM) has been shown to induce female germ cell development from embryonic stem cells (ESCs). Testicular cells (TCs) secrete a...
BACKGROUND
Testicular cell conditioned medium (TCCM) has been shown to induce female germ cell development from embryonic stem cells (ESCs). Testicular cells (TCs) secrete a variety of growth factors such as growth differentiation factor-9 (GDF-9), bone morphogenetic protein 4 (BMP-4), stem cell factor (SCF), leukemia inhibitory factor (LIF), and other, that could improve oocyte maturation. Here we have investigated the effect of human TCCM (hTCCM) on maturation (IVM) and morphology of mouse oocytes.
MATERIALS AND METHODS
In this experimental study, 360 germinal vesicle (GV) oocytes were obtained from NMRI mice, aged 4-6 weeks that had received 5 IU pregnant mare's serum gonadotropin (PMSG) 48 hours before. GV oocytes were subjected to IVM. 120 GV oocytes were cultured in each medium; hTCCM as the test group, DMEM + 20%FBS as the control group and Ham's F10 + HFF medium as the sham group. The rates of the IVM and perivitelline space (PVS) changes were recorded at 8, 16 and 24 hours after culture. The metaphase II (MII) oocytes were subjected for fertilization (IVF) and the fertilization rate was evaluated after 1, 2, and 3 days.
RESULTS
There was a significant difference between the maturation rates in hTCCM (31.67% MII) and the control [0% MII, P<0.05, (7.5% MI, 52.5% deg. and 40%GV)] groups but there was not a significant difference between the maturation rates in hTCCM and the sham group (53.33% MII, P>0.05). IVF success rate for MII oocytes obtained from IVM in the hTCCM group was 28.94% (n=11). Our data showed that hTCCM is an effective medium for GV oocyte growth and maturation compared to the control medium.
CONCLUSION
Our findings show that TCCM supports oocyte IVM in mice and affect oocyte morphology.
PubMed: 33098383
DOI: 10.22074/ijfs.2020.6097 -
Journal of Assisted Reproduction and... Jan 2022Does existing scientific literature suggest an impact of oocyte dysmorphisms on biological or clinical outcomes of assisted reproduction treatments?
PURPOSE
Does existing scientific literature suggest an impact of oocyte dysmorphisms on biological or clinical outcomes of assisted reproduction treatments?
METHODS
Studies of interest were selected from an initial cohort of 6651 potentially relevant records retrieved. PubMed was systematically searched for peer-reviewed original papers and reviews identified by keywords and medical subject heading (MeSH) terms. The most relevant publications were critically evaluated to identify criteria for oocyte morphological evaluation and IVF outcomes. For each morphological abnormality, we generated an oocyte literature score (OLS) through the following procedure: (a) papers showing a negative, absence of, or positive correlation between a given abnormality and IVF outcome were scored 1, 0, and - 1, respectively; (b) the sum of these scores was expressed as a fraction of all analyzed papers; (c) the obtained fraction was multiplied by 10 and converted into decimal number.
RESULT
We identified eleven different dysmorphisms, of which six were extracytoplasmic (COC, zona pellucida, perivitelline space, polar body 1, shape, giant size) and five intracytoplasmic (vacuoles, refractile bodies, SER clusters, granularity, color). Among the extracytoplasmic dysmorphisms, abnormal morphology of the COC generated an OLS of 8.33, indicating a large prevalence (5/6) of studies associated with a negative outcome. Three intracytoplasmic dysmorphisms (vacuoles, SER clusters, and granularity) produced OLS of 7.14, 7.78, and 6.25, respectively, suggestive of a majority of studies reporting a negative outcome.
CONCLUSION
COC morphology, vacuoles, SER clusters, and granularity produced OLS suggestive of a prevalence of studies reporting a negative outcome.
Topics: Humans; Oocytes; Oogenesis; Zona Pellucida
PubMed: 34993709
DOI: 10.1007/s10815-021-02370-3 -
The Journal of Reproduction and... Oct 2018Previously, our group demonstrated that recombinant porcine oviductin (pOVGP1) binds to the zona pellucida (ZP) of in vitro-matured (IVM) porcine oocytes with a positive...
Previously, our group demonstrated that recombinant porcine oviductin (pOVGP1) binds to the zona pellucida (ZP) of in vitro-matured (IVM) porcine oocytes with a positive effect on in vitro fertilization (IVF). The fact that pOVGP1 was detected inside IVM oocytes suggested that this protein had a biological role during embryo development. The aim of this study was to evaluate the effects of pOVGP1 on bovine in vitro embryo development. We applied 10 or 50 µg/ml of pOVGP1 during IVF, embryonic in vitro culture (IVC), or both, to evaluate cleavage and embryo development. Blastocyst quality was assessed by analyzing the expression of important developmental genes and the survival rates after vitrification/warming. pOVGP1 was detected in the ZP, perivitelline space, and plasma membrane of blastocysts. No significant differences (P > 0.05) were found in cleavage or blastocyst yield when 10 or 50 µg/ml of pOVGP1 was used during IVF or IVC. However, when 50 µg/ml pOVGP1 was used during IVF + IVC, the number of blastocysts obtained was half that obtained with the control and 10 µg/ml pOVGP1 groups. The survival rates after vitrification/warming of expanded blastocysts cultured with pOVGP1 showed no significant differences between groups (P > 0.05). The use of pOVGP1 during IVF, IVC, or both, increased the relative abundance of mRNA of DSC2, ATF4, AQP3, and DNMT3A, the marker-genes of embryo quality. In conclusion, the use of pOVGP1 during bovine embryo in vitro culture does not affect embryo developmental rates but produces embryos of better quality in terms of the relative abundance of specific genes.
Topics: Animals; Animals, Genetically Modified; Blastocyst; Cattle; Cell Membrane; Embryo Transfer; Embryonic Development; Female; Fertilization in Vitro; Glycoproteins; Male; Microscopy, Confocal; Oocytes; Oviducts; Recombinant Proteins; Spermatozoa; Swine; Vitrification; Zona Pellucida
PubMed: 30078833
DOI: 10.1262/jrd.2018-058 -
International Journal of Molecular... Apr 2017In animals, hatching represents the transition point from a developing embryo to a free-living individual, the larva. This process is finely regulated by many endogenous...
In animals, hatching represents the transition point from a developing embryo to a free-living individual, the larva. This process is finely regulated by many endogenous and environmental factors and has been shown to be sensitive to a variety of chemical agents. It is commonly evaluated in bioassays in order to establish the effects of different agents on early development and reproductive capabilities in fish and other aquatic animals. In fish, the breakdown of the chorion is achieved by the secretion of choriolysin by hatching gland cells (HGCs) into the perivitelline space (PVS), coupled with spontaneous movements of the developing larva. In this work, we used zebrafish to assay the effects of a family of widely used agrochemicals-triazoles Triadimefon (FON), Triadimenol (NOL) and free triazole (1,2,4-T)-on hatching success. We found a strong inhibition of hatching by triazole exposure which was correlated with morphological changes and a reduction in the secretory function of the HGCs. As a consequence, the release of choriolytic enzymes by HGCs was reduced. We also found that HGC secretion reduction after exposure to FON can be rescued by co-incubation with a dopamine D2 receptor antagonist but not by antagonists of the D1-like receptors. This suggests a specific pathway through which this family of fungicides may be impairing a critical event in the fish life cycle.
Topics: Animals; Biological Assay; Ecotoxicology; Embryo, Nonmammalian; Fungicides, Industrial; Inhibitory Concentration 50; Larva; Motor Activity; Peptide Hydrolases; Proteolysis; Triazoles; Zebrafish; Zebrafish Proteins
PubMed: 28375163
DOI: 10.3390/ijms18040710