The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditi...The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditions during the first ten days of incubation at high altitudes on Leghorn hens hatching eggs. Five hundred four hatching eggs were equally divided into three treatment groups and placed in twelve incubators (R = 4). The first group was subjected to standard ventilated conditions (V) during the setting phase. The ventilation inlet holes of the remaining incubators in the NV treatments were closed with either micropore (M) or polypropylene (P) tape, referred to as NVM and NVP groups, respectively. These two different airtight settings were intended to allow for a gradual rise in CO2 naturally generated by the embryos. Results indicate that carbon dioxide concentration gradually increased during the first half of incubation, reaching 1.42% in the NVM group and 1.20% in the NVP group, while the V condition group remained at 0.15%. From 10 days of incubation onwards, normal V conditions were restored in all incubators. The highest hatchability of fertile eggs (HFE) was shown by the NVP group (55.7%), followed by the V (52.6%) and NVM (38.6%) groups. The NVP group showed a greater yolk-free body mass (YFBM) from 10 days of incubation until the hatch basket transfer. NV conditions during the first 10 days of incubation at high altitude produced higher YFBM with gradually decreasing yolk sac mass. In comparison to the NVM and V conditions, the particular NVP condition showed a beneficial impact on the quality of hatched chicks. Sustaining NVP condition (1.2% of CO2) throughout the first half of incubation at high altitude generated the optimal environment in the incubator ensuring the best hatchability results. This study highlights how important it is for hatchery managers to recognize the influence of low O2 and high levels of CO2 on the development trajectories of Leghorn embryos during early incubation at high altitudes.展开更多
Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,...Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,we performed genomic sequencing for 80 core maize germplasms and constructed a high-density genomic variation map using our newly developed pipeline(MQ2Gpipe).Based on the induction rate of EC(REC),these inbred lines were categorized into three subpopulations.The low-REC germplasms displayed more abundant genetic diversity than the high-REC germplasms.By integrating a genome-wide selective signature screen and region-based association analysis,we revealed 95.23 Mb of selective regions and 43 REC-associated variants.These variants had phenotypic variance explained values ranging between 21.46 and 49.46%.In total,103 candidate genes were identified within the linkage disequilibrium regions of these REC-associated loci.These genes mainly participate in regulation of the cell cycle,regulation of cytokinesis,and other functions,among which MYB15 and EMB2745 were located within the previously reported QTL for EC induction.Numerous leaf area-associated variants with large effects were closely linked to several REC-related loci,implying a potential synergistic selection of REC and leaf size during modern maize breeding.展开更多
Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficienc...Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficiency is limited. For this purpose, the embryonic and larval development of reciprocal crosses of Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) were studied under laboratory conditions. The fertilization rate varied from 63.33% to 92%, while the hatching rate ranged from 55.68% to 76% with the highest value in hybrids ♀Cg × ♂Cj. Crosses between ♀Cj × ♂Cj, ♀Cg × ♂Cj and ♀Cj × ♂Cg had embryonic stages similar to those of the pure sib ♀Cg x ♂Cg. All crosses, however, had different timing for the various embryological stages. Hatching occurred at 32 h 15 min and 38 h for ♀Cj × ♂Cj and ♀Cj × ♂Cg, and 23 h and 23 h 30 min, respectively, for ♀Cg × ♂Cg and ♀Cg × ♂Cj. However, both crosses produced viable larvae until the first external feeding. The external morphological features of the larvae were completely formed by the 10th day after hatching. The body forms of the crosses at this time were indistinguishable from the pure sib. This study thus laid the groundwork for further comparative studies on hybrid performance and characterization.展开更多
BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure....BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.展开更多
BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage embryos.ESCs exhibit a unique combination of self-renewal potency,...BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage embryos.ESCs exhibit a unique combination of self-renewal potency,unlimited proliferation,and pluripotency.The latter is evident by the ability of the isolated cells to differ-entiate spontaneously into multiple cell lineages,representing the three primary embryonic germ layers.Multiple regulatory networks guide ESCs,directing their self-renewal and lineage-specific differentiation.Apoptosis,or programmed cell death,emerges as a key event involved in sculpting and forming various organs and structures ensuring proper embryonic development.How-ever,the molecular mechanisms underlying the dynamic interplay between diffe-rentiation and apoptosis remain poorly understood.AIM To investigate the regulatory impact of apoptosis on the early differentiation of ESCs into cardiac cells,using mouse ESC(mESC)models-mESC-B-cell lym-phoma 2(BCL-2),mESC-PIM-2,and mESC-metallothionein-1(MET-1)-which overexpress the anti-apoptotic genes Bcl-2,Pim-2,and Met-1,respectively.METHODS mESC-T2(wild-type),mESC-BCL-2,mESC-PIM-2,and mESC-MET-1 have been used to assess the effect of potentiated apoptotic signals on cardiac differentiation.The hanging drop method was adopted to generate embryoid bodies(EBs)and induce terminal differentiation of mESCs.The size of the generated EBs was measured in each condition compared to the wild type.At the functional level,the percentage of cardiac differentiation was measured by calculating the number of beating cardiomyocytes in the manipulated mESCs compared to the control.At the molecular level,quantitative reverse transcription-polymerase chain reaction was used to assess the mRNA expression of three cardiac markers:Troponin T,GATA4,and NKX2.5.Additionally,troponin T protein expression was evaluated through immunofluorescence and western blot assays.RESULTS Our findings showed that the upregulation of Bcl-2,Pim-2,and Met-1 genes led to a reduction in the size of the EBs derived from the manipulated mESCs,in comparison with their wild-type counterpart.Additionally,a decrease in the count of beating cardiomyocytes among differentiated cells was observed.Furthermore,the mRNA expression of three cardiac markers-troponin T,GATA4,and NKX2.5-was diminished in mESCs overexpressing the three anti-apoptotic genes compared to the control cell line.Moreover,the overexpression of the anti-apoptotic genes resulted in a reduction in troponin T protein expression.CONCLUSION Our findings revealed that the upregulation of Bcl-2,Pim-2,and Met-1 genes altered cardiac differentiation,providing insight into the intricate interplay between apoptosis and ESC fate determination.展开更多
We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota (Amphibia, Anura, Ranidae). Embryos were derived from artificial fertilization of frogs’ eggs, and the...We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota (Amphibia, Anura, Ranidae). Embryos were derived from artificial fertilization of frogs’ eggs, and the staging of development was based on morphological and physiological characteristics. Two major periods of development were designated: i) early embryonic period, from fertilization to operculum completion stage, lasted for 324 h at water temperature (WT) 18 ?23℃; ii) larval period, from operculum completion stage to tail absorbed stage, took 1207 h at WT 20 ? 24℃. Tadpoles of the concave-eared torrent frogs showed no evidence of abdominal sucker. Absence of this key characteristic supports the view from molecular systematics that concave-eared torrent frog does not belong to the genus Amolops. Two cleavage patterns were observed in embryos at 8-cell and 16-cell stages, with Pattern I2 (latitudinal cleavage at the 8-cell stage, and meridional cleavage at the 16-cell stage with two perpendicular meridional furrows) being the predominant pattern and only 1.5% belonging to Pattern II (meridional cleavage at the 8-cell stage and latitudinal cleavage at the 16-cell stage). The factors affecting cleavage and hatching ratios, developmental speed, and ecological adaptation were discussed.展开更多
Rhesus monkey embryonic stem(rES) cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,wh...Rhesus monkey embryonic stem(rES) cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm(DE) cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins(CK7,CK18,CK19,CK20,and OV-6) and genes(GSTPi,IB4,and HNF1β).At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ~90% to ~20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.展开更多
[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with te...[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5. The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage. The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] It provided important reference for studying artificial propagation and genetic breeding of loach.展开更多
[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down ...[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.展开更多
[Objective] The research aimed to provide technical basis for fry rearing of Silnrus asotus in Dongting Lake.[Method] The induced spawning medicine was used in the experiment to conduct artificial induced spawning and...[Objective] The research aimed to provide technical basis for fry rearing of Silnrus asotus in Dongting Lake.[Method] The induced spawning medicine was used in the experiment to conduct artificial induced spawning and fertilization for obtaining round green fertilized eggs.According to embryonic development,the morphological characteristics of embryo at different developmental stages were recorded detailedly through microscope.[Result] The embryonic development of Silnrus asotus in Dongting Lake was divided into 7 stages,namely, blastoderm stage,cleavage stage, blastula stage, gastrula stage,neurula stage,organogenesis stage and pre-hatching stage.After hatched for 37 h 20 min in water at 22-24 ℃, fries were come out.[Conclusion] The time sequence of Silnrus asotus in Dongting Lake was basically similar to that of other catfish,while its hatching time was shorter than that of other fish in Siluriformes.展开更多
The localization of four neuropeptidelike substances during embryonic development in amphibian was studied by using immuno cytochemical technique. The cells with positive reaction appeared firstly in the endoderm ce...The localization of four neuropeptidelike substances during embryonic development in amphibian was studied by using immuno cytochemical technique. The cells with positive reaction appeared firstly in the endoderm cells during early tailbud stage, and then were detected in connective tissue at the outer portion of gastrointestinal tract during tadpole stage. In the nervous system, the cells with positive reaction were observed in cranial ganglion and glial cells at the outer margin of the brain and in the inner wall of ventricles. They were also frequently observed in the epidermis during late tailbud stage. The relationship between the appearance of neuropeptides in timespatial sequences and the development of nervous system, the neural crest origin of the cells with positive reaction, and the role of epidermal conductivity in neuropeptidelike cells in epiderms were discussed.展开更多
Embryonic development was studied in six cross combinations ofCitrus sinensis x C. tangerina, C. sinensis x C. reticulata, C. sinensis x (C. tangerina + C.reticulata), C. sinensis x Poncirus trifoliate, C.reticulata x...Embryonic development was studied in six cross combinations ofCitrus sinensis x C. tangerina, C. sinensis x C. reticulata, C. sinensis x (C. tangerina + C.reticulata), C. sinensis x Poncirus trifoliate, C.reticulata x C grandis and C. grandis xPoncirus trifoliate. The results showed that on the 30th day after pollination thezygote remained a single cell. On the 40th day, the zygote began to divide. On the50th day, zygotic embryo became globular-shaped while nucellar embryos had notinvaded the embryo sac. On the 55th day, a few nucellar embryos began to invadethe embryo sac. On the 60th day, the zygotic embryo became heart-shaped, and atthe same time, a large number of nucellar embryos invaded the embryo sac. On the80th day after pollination, the zygotic embryo was surrounded by nucellar embryosand it was not easy to distinguish these embryos. The cross combination affected thedevelopments of zygotic embryos, ovules and fruits, which were mainly determined bythe cross parents. As compared with interspecies crossing, the zygotic division ofintergenus crossing began later, the zygotic embryos developed slowlier and theinvading time of nucellar embryos was also delayed.展开更多
[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for i...[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for intensive breeding and propagation by artificial insemination. The entire embryonic development process of L. crassilabris from insemination to hatching out of larvae fish was observed consecutively. [Result] Fertilized eggs of L. crassilabris are spherical, and the egg diameter is about 2.2-2.4 mm after water absorption; under conditions of water temperature ranging from 27 to 28 ℃, the embryonic development of L. crassilabris from insemination to hatching out of larvae fish lasted 2 496 min, with a total accumulated temperature of 1 123-1 165 h·℃. [Conclusion] This study is advantageous to better understand the characteristics of embryonic development of L. crassilabris and provides basic biological data for protection and utilization of fish resources and other related work.展开更多
Metabolism of hydrogen peroxide in the course of embryonic development ofsilkworm (variety Guang) was determined by using colorimetric analysis and oxygen electrodemethod. The results indicated that: 1) In the course ...Metabolism of hydrogen peroxide in the course of embryonic development ofsilkworm (variety Guang) was determined by using colorimetric analysis and oxygen electrodemethod. The results indicated that: 1) In the course of fertilization (0-4 h after egg laying), thelevel of H_2O_2 content reached its peak value at 2.5 h of the developmental course and the activity ofsuperoxidase dismutase (SOD) was at higher level while the activity of catalase (CAT) at the lowestcorrespondingly; 2) The H_2O_2 content in embryo, in which the diapause of eggs was being relievedthrough treatment with hydrochloric acid on time in the course of embryonic development, wassignificantly higher than that of diapause eggs except the lower level showed in the embryo whenthe development of it went on for 168~216 h and the activity of SOD reached, lower and higher,tWo peaks in 72 and 168 h after egg-laying, respectively, and was significantly higher in late stagewhile the activity of CAT was shown with a stable level in the period of 72-192 h after egg-laying,and, after then, a rapid rising occurred in the embryo. The level of the CAT activity in embryowas shown significantly lower than that in diapause eggs in early period and higher in late stage ofegg development; 3) In the course of formation of diapause in eggs, the level of H_2O_2contentchanged smoothly and the activity of SOD changed vigorously in early period, and kept stable statelater; and the CAT activity increased gradually; while in the course of relief of diapause under ontime-treatment with hydrochloric acid, the level of H_2O_2 was significantly higher than that indiapause eggs and the SOD activity displayed a new peak value and significantly rose in later stage,while the activity of CAT in relieving embryo from diapause was signincantly lower than that indiapause eggs. Combining the results obtained in other researches with those from ours mentionedabove, we suggest that the metabolism of H_2O_2 might be of importance in the courses of formationand relief of diapause in silkworm eggs. Maybe the esterase A4 timer hypothesis and themicropylar barrier to oxygen hypothesis could be integrated for explanation of the course offormation and relief of diapause in silkworm eggs.展开更多
mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the ...mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.展开更多
[Objective] This study aimed to observe the embryonic development of the F1 hybrid between Ryukin (♀) and Dragon-eye (♂). [Method] Embryonic development of the FI hybrid and parents between Ryukin (♀) and Dra...[Objective] This study aimed to observe the embryonic development of the F1 hybrid between Ryukin (♀) and Dragon-eye (♂). [Method] Embryonic development of the FI hybrid and parents between Ryukin (♀) and Dragon-eye (♂) was observed with XTL-2400 anatomical lens continuously. The organ characteristics and structures at each development stage were observed with OLYMPUS CKX41 microscopes, and photographed and recorded with a digital camera. [Result] At a water temperature of 25℃, the hatching time of the F1 hybrid is 51.3 h, the hatching time of Ryukin embryo is 52.4 h and that of Dragon-eye embryo is 56.3 h. [Conclusion] This study provides certain theoretical and practical basis for improving morphological characteristics of the existing Dragon-eye goldfish.展开更多
文摘The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditions during the first ten days of incubation at high altitudes on Leghorn hens hatching eggs. Five hundred four hatching eggs were equally divided into three treatment groups and placed in twelve incubators (R = 4). The first group was subjected to standard ventilated conditions (V) during the setting phase. The ventilation inlet holes of the remaining incubators in the NV treatments were closed with either micropore (M) or polypropylene (P) tape, referred to as NVM and NVP groups, respectively. These two different airtight settings were intended to allow for a gradual rise in CO2 naturally generated by the embryos. Results indicate that carbon dioxide concentration gradually increased during the first half of incubation, reaching 1.42% in the NVM group and 1.20% in the NVP group, while the V condition group remained at 0.15%. From 10 days of incubation onwards, normal V conditions were restored in all incubators. The highest hatchability of fertile eggs (HFE) was shown by the NVP group (55.7%), followed by the V (52.6%) and NVM (38.6%) groups. The NVP group showed a greater yolk-free body mass (YFBM) from 10 days of incubation until the hatch basket transfer. NV conditions during the first 10 days of incubation at high altitude produced higher YFBM with gradually decreasing yolk sac mass. In comparison to the NVM and V conditions, the particular NVP condition showed a beneficial impact on the quality of hatched chicks. Sustaining NVP condition (1.2% of CO2) throughout the first half of incubation at high altitude generated the optimal environment in the incubator ensuring the best hatchability results. This study highlights how important it is for hatchery managers to recognize the influence of low O2 and high levels of CO2 on the development trajectories of Leghorn embryos during early incubation at high altitudes.
基金supported by the National Key Research and Development Program of China(2021YFF1000303)the National Nature Science Foundation of China(32072073,32001500,and 32101777)the Sichuan Science and Technology Program,China(2021JDTD0004 and 2021YJ0476)。
文摘Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,we performed genomic sequencing for 80 core maize germplasms and constructed a high-density genomic variation map using our newly developed pipeline(MQ2Gpipe).Based on the induction rate of EC(REC),these inbred lines were categorized into three subpopulations.The low-REC germplasms displayed more abundant genetic diversity than the high-REC germplasms.By integrating a genome-wide selective signature screen and region-based association analysis,we revealed 95.23 Mb of selective regions and 43 REC-associated variants.These variants had phenotypic variance explained values ranging between 21.46 and 49.46%.In total,103 candidate genes were identified within the linkage disequilibrium regions of these REC-associated loci.These genes mainly participate in regulation of the cell cycle,regulation of cytokinesis,and other functions,among which MYB15 and EMB2745 were located within the previously reported QTL for EC induction.Numerous leaf area-associated variants with large effects were closely linked to several REC-related loci,implying a potential synergistic selection of REC and leaf size during modern maize breeding.
文摘Establishing intraspecific breeding and hybridization programs and determining genetic variability are two important issues for aquaculture. However, interspecific hybridization to improve growth and feeding efficiency is limited. For this purpose, the embryonic and larval development of reciprocal crosses of Clarias gariepinus (Burchell, 1822) and Clarias jaensis (Boulenger, 1909) were studied under laboratory conditions. The fertilization rate varied from 63.33% to 92%, while the hatching rate ranged from 55.68% to 76% with the highest value in hybrids ♀Cg × ♂Cj. Crosses between ♀Cj × ♂Cj, ♀Cg × ♂Cj and ♀Cj × ♂Cg had embryonic stages similar to those of the pure sib ♀Cg x ♂Cg. All crosses, however, had different timing for the various embryological stages. Hatching occurred at 32 h 15 min and 38 h for ♀Cj × ♂Cj and ♀Cj × ♂Cg, and 23 h and 23 h 30 min, respectively, for ♀Cg × ♂Cg and ♀Cg × ♂Cj. However, both crosses produced viable larvae until the first external feeding. The external morphological features of the larvae were completely formed by the 10th day after hatching. The body forms of the crosses at this time were indistinguishable from the pure sib. This study thus laid the groundwork for further comparative studies on hybrid performance and characterization.
基金Supported by the Shandong Provincial Traditional Chinese Medicine Science and Technology Development Program,No.C-262the 2021 Science and Technology Innovation Research Project of Shandong Maternal and Child Health Association,No.2021-19-24.
文摘BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.
基金Supported by the National Council for Scientific Research in Lebanon,CNRS-L.
文摘BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage embryos.ESCs exhibit a unique combination of self-renewal potency,unlimited proliferation,and pluripotency.The latter is evident by the ability of the isolated cells to differ-entiate spontaneously into multiple cell lineages,representing the three primary embryonic germ layers.Multiple regulatory networks guide ESCs,directing their self-renewal and lineage-specific differentiation.Apoptosis,or programmed cell death,emerges as a key event involved in sculpting and forming various organs and structures ensuring proper embryonic development.How-ever,the molecular mechanisms underlying the dynamic interplay between diffe-rentiation and apoptosis remain poorly understood.AIM To investigate the regulatory impact of apoptosis on the early differentiation of ESCs into cardiac cells,using mouse ESC(mESC)models-mESC-B-cell lym-phoma 2(BCL-2),mESC-PIM-2,and mESC-metallothionein-1(MET-1)-which overexpress the anti-apoptotic genes Bcl-2,Pim-2,and Met-1,respectively.METHODS mESC-T2(wild-type),mESC-BCL-2,mESC-PIM-2,and mESC-MET-1 have been used to assess the effect of potentiated apoptotic signals on cardiac differentiation.The hanging drop method was adopted to generate embryoid bodies(EBs)and induce terminal differentiation of mESCs.The size of the generated EBs was measured in each condition compared to the wild type.At the functional level,the percentage of cardiac differentiation was measured by calculating the number of beating cardiomyocytes in the manipulated mESCs compared to the control.At the molecular level,quantitative reverse transcription-polymerase chain reaction was used to assess the mRNA expression of three cardiac markers:Troponin T,GATA4,and NKX2.5.Additionally,troponin T protein expression was evaluated through immunofluorescence and western blot assays.RESULTS Our findings showed that the upregulation of Bcl-2,Pim-2,and Met-1 genes led to a reduction in the size of the EBs derived from the manipulated mESCs,in comparison with their wild-type counterpart.Additionally,a decrease in the count of beating cardiomyocytes among differentiated cells was observed.Furthermore,the mRNA expression of three cardiac markers-troponin T,GATA4,and NKX2.5-was diminished in mESCs overexpressing the three anti-apoptotic genes compared to the control cell line.Moreover,the overexpression of the anti-apoptotic genes resulted in a reduction in troponin T protein expression.CONCLUSION Our findings revealed that the upregulation of Bcl-2,Pim-2,and Met-1 genes altered cardiac differentiation,providing insight into the intricate interplay between apoptosis and ESC fate determination.
基金supported by the National Natural Science Fundation of China(30730029)
文摘We investigated the early embryonic and larval development of the concave-eared torrent frogs, Odorrana tormota (Amphibia, Anura, Ranidae). Embryos were derived from artificial fertilization of frogs’ eggs, and the staging of development was based on morphological and physiological characteristics. Two major periods of development were designated: i) early embryonic period, from fertilization to operculum completion stage, lasted for 324 h at water temperature (WT) 18 ?23℃; ii) larval period, from operculum completion stage to tail absorbed stage, took 1207 h at WT 20 ? 24℃. Tadpoles of the concave-eared torrent frogs showed no evidence of abdominal sucker. Absence of this key characteristic supports the view from molecular systematics that concave-eared torrent frog does not belong to the genus Amolops. Two cleavage patterns were observed in embryos at 8-cell and 16-cell stages, with Pattern I2 (latitudinal cleavage at the 8-cell stage, and meridional cleavage at the 16-cell stage with two perpendicular meridional furrows) being the predominant pattern and only 1.5% belonging to Pattern II (meridional cleavage at the 8-cell stage and latitudinal cleavage at the 16-cell stage). The factors affecting cleavage and hatching ratios, developmental speed, and ecological adaptation were discussed.
基金supported by research grants from Zhejiang Natural Sciences Foundation of China (Y2110911 Y2080996)the National Key Technologies R&D Program of China (2007CB947701)
文摘Rhesus monkey embryonic stem(rES) cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm(DE) cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins(CK7,CK18,CK19,CK20,and OV-6) and genes(GSTPi,IB4,and HNF1β).At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ~90% to ~20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.
文摘[Objective] Aim to know the whole process of embryonic development of loach. [Method] DOM + LHRH-A2 was used to induce spawning of loach,then after fertilization,the embryos were cultured into freshwater water with temperature from 24 to 26 ℃ and pH from 7.0 to 7.5. The embryonic development of loach was observed and 27 concrete morphological characteristics and development time of loach from fertilized egg to newly hatched larval period were described in detail. [Result] The embryonic development of loach could be divided into cleavage stage,blastocyst stage,gastrula stage,neurula stage and organogenesis stage. The loach embryo from fertilized egg to out membrane period was 30 h 45 min in fresh water from 24 to 26 ℃ and pH from 7.0 to 7.5. [Conclusion] It provided important reference for studying artificial propagation and genetic breeding of loach.
基金Supported by Project of Baotou University(BSY2010-23)~~
文摘[Objective] The paper was to establish embryonic stem cell system of goats. [Method] Numerous primordial germ cell colonies were derived from gonadal ridge and the surrounding tissues in 20 millimeter fetuses of down producing goat. Primordial germ cells and goats embryonic fibroblasts obtained from conceptus of equivaient gestational age were co-cultured. [Result] The colonies showed some characteristics of embryonic stem cells, such as the morphology of nest-like, they continued to be AKP positive and the ability to be continuously passed [Conclusion] These cells were pluripotent and ES-like cells.
文摘[Objective] The research aimed to provide technical basis for fry rearing of Silnrus asotus in Dongting Lake.[Method] The induced spawning medicine was used in the experiment to conduct artificial induced spawning and fertilization for obtaining round green fertilized eggs.According to embryonic development,the morphological characteristics of embryo at different developmental stages were recorded detailedly through microscope.[Result] The embryonic development of Silnrus asotus in Dongting Lake was divided into 7 stages,namely, blastoderm stage,cleavage stage, blastula stage, gastrula stage,neurula stage,organogenesis stage and pre-hatching stage.After hatched for 37 h 20 min in water at 22-24 ℃, fries were come out.[Conclusion] The time sequence of Silnrus asotus in Dongting Lake was basically similar to that of other catfish,while its hatching time was shorter than that of other fish in Siluriformes.
文摘The localization of four neuropeptidelike substances during embryonic development in amphibian was studied by using immuno cytochemical technique. The cells with positive reaction appeared firstly in the endoderm cells during early tailbud stage, and then were detected in connective tissue at the outer portion of gastrointestinal tract during tadpole stage. In the nervous system, the cells with positive reaction were observed in cranial ganglion and glial cells at the outer margin of the brain and in the inner wall of ventricles. They were also frequently observed in the epidermis during late tailbud stage. The relationship between the appearance of neuropeptides in timespatial sequences and the development of nervous system, the neural crest origin of the cells with positive reaction, and the role of epidermal conductivity in neuropeptidelike cells in epiderms were discussed.
文摘Embryonic development was studied in six cross combinations ofCitrus sinensis x C. tangerina, C. sinensis x C. reticulata, C. sinensis x (C. tangerina + C.reticulata), C. sinensis x Poncirus trifoliate, C.reticulata x C grandis and C. grandis xPoncirus trifoliate. The results showed that on the 30th day after pollination thezygote remained a single cell. On the 40th day, the zygote began to divide. On the50th day, zygotic embryo became globular-shaped while nucellar embryos had notinvaded the embryo sac. On the 55th day, a few nucellar embryos began to invadethe embryo sac. On the 60th day, the zygotic embryo became heart-shaped, and atthe same time, a large number of nucellar embryos invaded the embryo sac. On the80th day after pollination, the zygotic embryo was surrounded by nucellar embryosand it was not easy to distinguish these embryos. The cross combination affected thedevelopments of zygotic embryos, ovules and fruits, which were mainly determined bythe cross parents. As compared with interspecies crossing, the zygotic division ofintergenus crossing began later, the zygotic embryos developed slowlier and theinvading time of nucellar embryos was also delayed.
基金Supported by Science and Technology Support Program of Science and Technology Department of Jiangsu Province(BE2009335)~~
文摘[Objective] This study aimed to observe the embryonic development process of Leiocassis crassilabris. [Method] Wild L. crassilabris collected from the Nanjing section of the Yangtze River was used as parent fish for intensive breeding and propagation by artificial insemination. The entire embryonic development process of L. crassilabris from insemination to hatching out of larvae fish was observed consecutively. [Result] Fertilized eggs of L. crassilabris are spherical, and the egg diameter is about 2.2-2.4 mm after water absorption; under conditions of water temperature ranging from 27 to 28 ℃, the embryonic development of L. crassilabris from insemination to hatching out of larvae fish lasted 2 496 min, with a total accumulated temperature of 1 123-1 165 h·℃. [Conclusion] This study is advantageous to better understand the characteristics of embryonic development of L. crassilabris and provides basic biological data for protection and utilization of fish resources and other related work.
文摘Metabolism of hydrogen peroxide in the course of embryonic development ofsilkworm (variety Guang) was determined by using colorimetric analysis and oxygen electrodemethod. The results indicated that: 1) In the course of fertilization (0-4 h after egg laying), thelevel of H_2O_2 content reached its peak value at 2.5 h of the developmental course and the activity ofsuperoxidase dismutase (SOD) was at higher level while the activity of catalase (CAT) at the lowestcorrespondingly; 2) The H_2O_2 content in embryo, in which the diapause of eggs was being relievedthrough treatment with hydrochloric acid on time in the course of embryonic development, wassignificantly higher than that of diapause eggs except the lower level showed in the embryo whenthe development of it went on for 168~216 h and the activity of SOD reached, lower and higher,tWo peaks in 72 and 168 h after egg-laying, respectively, and was significantly higher in late stagewhile the activity of CAT was shown with a stable level in the period of 72-192 h after egg-laying,and, after then, a rapid rising occurred in the embryo. The level of the CAT activity in embryowas shown significantly lower than that in diapause eggs in early period and higher in late stage ofegg development; 3) In the course of formation of diapause in eggs, the level of H_2O_2contentchanged smoothly and the activity of SOD changed vigorously in early period, and kept stable statelater; and the CAT activity increased gradually; while in the course of relief of diapause under ontime-treatment with hydrochloric acid, the level of H_2O_2 was significantly higher than that indiapause eggs and the SOD activity displayed a new peak value and significantly rose in later stage,while the activity of CAT in relieving embryo from diapause was signincantly lower than that indiapause eggs. Combining the results obtained in other researches with those from ours mentionedabove, we suggest that the metabolism of H_2O_2 might be of importance in the courses of formationand relief of diapause in silkworm eggs. Maybe the esterase A4 timer hypothesis and themicropylar barrier to oxygen hypothesis could be integrated for explanation of the course offormation and relief of diapause in silkworm eggs.
文摘mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.
文摘[Objective] This study aimed to observe the embryonic development of the F1 hybrid between Ryukin (♀) and Dragon-eye (♂). [Method] Embryonic development of the FI hybrid and parents between Ryukin (♀) and Dragon-eye (♂) was observed with XTL-2400 anatomical lens continuously. The organ characteristics and structures at each development stage were observed with OLYMPUS CKX41 microscopes, and photographed and recorded with a digital camera. [Result] At a water temperature of 25℃, the hatching time of the F1 hybrid is 51.3 h, the hatching time of Ryukin embryo is 52.4 h and that of Dragon-eye embryo is 56.3 h. [Conclusion] This study provides certain theoretical and practical basis for improving morphological characteristics of the existing Dragon-eye goldfish.