Somite formation in cultured embryos of the snapping turtle, Chelydra serpentina

Development ◽  
1980 ◽  
Vol 59 (1) ◽  
pp. 113-130
Author(s):  
David S. Packard
Keyword(s):  
Neural Tube ◽  
In Vitro Cultivation ◽  
Slight Variation ◽  
Chelydra Serpentina ◽  
Tail Bud ◽  
Snapping Turtle ◽  
Wide Range ◽  
Somite Formation ◽  
Full Complement

A simple, reliable method for the in vitro cultivation of snapping turtle embryos was demonstrated. This technique was used to study somite formation in explants containing segmental plates. Segmental plates formed a full complement of somites whether the neural tube or the neural tube and notochord was present. Explanted snapping turtle segmental plates formed an average of 6·6 ± 1·2 somites. Removal of the node region or tail bud from cultured intact embryos led to a cessation of somite formation after an additional 6·1 ± 1·8 somites had formed. These results indicate the number of somites the snapping turtle segmental plate will form. Also, the number of somites formed by explanted segmental plates showed only slight variation over a wide range of segmental plate lengths. It was concluded that while snapping turtle segmental plates formed significantly fewer somites than chicken or Japanese quail segmental plates, they were similar to the avian explants in their ability to form a consistent number of somites regardless of the length of the segmental plate.

scholarly journals In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review

2012 ◽  
Vol 21 (2) ◽  
pp. 81-86 ◽  
Author(s):  
Lygia Maria Friche Passos
Keyword(s):  
Cell Lines ◽  
Anaplasma Marginale ◽  
In Vitro Cultivation ◽  
Continuous Cell Lines ◽  
Wide Range ◽  
Antigenic Material ◽  
Subsequent Propagation ◽  
Species Specific

Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens, Anaplasma marginale and Anaplasma phagocytophilum. These in vitro cultivation systems have been used in a wide range of studies, covering morphological ultrastructural analysis, genetics, proteomics and biological differences between strains, including genome transcriptional and protein expression approaches, enabling comparisons between host and vector cells. Thus, such systems open a new window for a better understanding of interactions between pathogens and tick cells. Last but not least, such systems contribute to the reduction in usage of animals for experimental research, as antigenic material can be produced in reasonably large quantities without the use of in vivo species-specific systems.

Morphological and experimental studies of the somitomeric organization of the segmental plate in snapping turtle embryos

Development ◽  
1984 ◽  
Vol 84 (1) ◽  
pp. 35-48
Author(s):  
David S. Packard ◽  
Stephen Meier
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Final Stage ◽  
Experimental Studies ◽  
The Other ◽  
Chelydra Serpentina ◽  
Microscopy Imaging ◽  
Snapping Turtle ◽  
Somite Formation ◽  
Scanning Electron

The segmental plate mesoderm of snapping turtle embryos (Chelydra serpentina) was examined with stereoscanning electron microscopy imaging. A metameric pattern was detected along the entire length of the segmental plates. This pattern consisted of a tandem sequence of mesodermal units, called somitomeres. Each somitomere was oval to cubic in shape and the processes of the constituent mesodermal cells tended to be arranged in concentric rings about the centre of the somitomere. Several experiments from a previous study (Packard, 1980b) of snapping turtle segmental plates were repeated, but, instead of culturing the explants and observing the numbers of somites that formed, the explants were fixed immediately for scanning electron microscopy and the number of somitomeres was counted. The segmental plates were found to contain an average of 6·5 ± 0·7 somitomeres, which is almost identical to the average number of somites formed by such segmental plates when cultured (6·6 ± 1·2). Furthermore, the number of somitomeres was identical in right and left explants removed from the same embryo, and the number of somitomeres was consistent regardless of the length of the segmental plate. Both of these observations are identical to those made previously for somite formation in culture. This association between numbers of somitomeres and somites strongly suggests that one gives rise to the other. Finally, it was demonstrated that for each somite formed by a segmental plate in culture, the segmental plate contained one less somitomere. This showed in a direct manner that turtle somitomeres become somites. It was concluded that the segmental plate mesoderm of snapping turtle embryos is already segmented, and that the ‘segmentation’ seen under a dissecting microscope is actually the final stage of somitomere differentiation into an epithelial somite.

scholarly journals Sphere-Forming Culture for Expanding Genetically Distinct Patient-Derived Glioma Stem Cells by Cellular Growth Rate Screening

Cancers ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 549
Author(s):  
Kayoung Shin ◽  
Hyemi Shin ◽  
Hee Jin Cho ◽  
Hyunju Kang ◽  
Jin-Ku Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factor ◽  
Growth Rates ◽  
Isocitrate Dehydrogenase ◽  
Cellular Growth ◽  
Glioma Stem Cells ◽  
In Vitro Cultivation ◽  
Isocitrate Dehydrogenase 1 ◽  
Wide Range

Diffusely infiltrating gliomas (DIGs) are difficult to completely resect and are associated with a high rate of tumor relapse and progression from low- to high-grade glioma. In particular, optimized short-term culture-enriching patient-derived glioma stem cells (GSCs) are essential for customizing the therapeutic strategy based on clinically feasible in vitro drug screening for a wide range of DIGs, owing to the high inter-tumoral heterogeneity. Herein, we constructed a novel high-throughput culture condition screening platform called ‘GFSCAN’, which evaluated the cellular growth rates of GSCs for each DIG sample in 132 serum-free combinations, using 13 previously reported growth factors closely associated with glioma aggressiveness. In total, 72 patient-derived GSCs with available genomic profiles were tested in GFSCAN to explore the association between cellular growth rates in specific growth factor combinations and genomic/molecular backgrounds, including isocitrate dehydrogenase 1 (IDH1) mutation, chromosome arm 1p and 19q co-deletion, ATRX chromatin remodeler alteration, and transcriptional subtype. GSCs were clustered according to the dependency on epidermal growth factor and basic fibroblast growth factor (E&F), and isocitrate dehydrogenase 1 (IDH1) wild-type GSCs showed higher E&F dependencies than IDH1 mutant GSCs. More importantly, we elucidated optimal combinations for IDH1 mutant glioblastoma and lower grade glioma GSCs with low dependencies on E&F, which could be an aid in clinical decision-making for these DIGs. Thus, we demonstrated the utility of GFSCAN in personalizing in vitro cultivation to nominate personalized therapeutic options, in a clinically relevant time frame, for individual DIG patients, where standard clinical options have been exhausted.

scholarly journals Study of Phytophthora infestans Mont. de Bary isolates in the planting of potatoes

2021 ◽  
pp. 86-91
Author(s):  
N. V. Matsishina ◽  
P. V. Fisenko ◽  
O. A. Sobko ◽  
I. V. Kim ◽  
D. I. Volkov ◽  
...  
Keyword(s):  
Phytophthora Infestans ◽  
Molecular Genetic ◽  
Microscopic Analysis ◽  
Molecular Genetic Analysis ◽  
In Vitro Cultivation ◽  
Genetic Characteristics ◽  
Potato Varieties ◽  
Phytotoxic Activity ◽  
Wide Range

Relevance. One of the most common diseases of potatoes and other nightshade family species is late blight caused by a pathogenic oomycete of the Phytophthora infestans (Mont.) de Bary. At least 100 species of phytophthora have been described in nature, affecting a wide range of plant species. The phytophthora population is heterogeneous and is represented by races, as well as different types of mating. This leads to a rapid adaptation of the pathogen and the emergence of new, more aggressive, and resistant races. Phytophthora is a parasite, the damage from which cannot be avoided within the organic farming framework. Therefore, it is particularly important to know the pathogenesis and racial composition of phytophthora in each individual region of Solanaceae cultivation.Research methodology. Differentiation and collection of material from the natural population were carried out using potato varieties with known R-genes in the genome. Isolation and introduction into the culture were carried out from leaves with the dampening chambers method, followed by cultivation on nutrient media. The pathogen was identified by microscopic analysis. Culture filtrates were obtained on the liquid nutritious medium, followed by liquid filtration and autoclaving. Phytotoxic activity was determined by the effect on the seedlings of the nightshade, grass, and pea families by the standard method. Molecular genetic analysis of the isolates was carried out by ISSR analysis; the primer, amplification mixture, and temperature profile of the reaction were selected according to the literature data; the calculation of genetic characteristics was carried out using POPGENE software packages.Results. Samples of seven Phytophthora infestans isolates were collected and introduced into culture. As a result of in vitro cultivation, morphological differences were revealed, expressed in the structure and color of the mycelium, the shape of the colonies, the nature of sporulation, the color of the reverse, and the medium under the colonies. The genetic differences of the natural phytophthora material introduced into the culture, collected from potato varieties with single resistance genes (R1, R3, R4), were revealed. Differences in the phytotoxic activity of the studied isolates' cultural filtrates were revealed. The isolated isolates demonstrate differentiation at the phenotypic, genetic and physiological levels, which allows us to speak about their belonging to races.

scholarly journals The northernmost occurrence of Chelydra serpentina in the eastern US during the Pleistocene

2016 ◽  
Author(s):  
Chase Brownstein
Keyword(s):  
New Jersey ◽  
North America ◽  
Late Pleistocene ◽  
Ice Sheet ◽  
Eastern North America ◽  
Chelydra Serpentina ◽  
Turtle Species ◽  
Snapping Turtle ◽  
Caudal Vertebrae ◽  
Wide Range

The snapping turtle species Chelydra serpentina, which has a wide range across North America, is extremely tolerant to cold and even freezing conditions. Here, I describe a single caudal vertebrae referred to Chelydra serpentina from the Late Pleistocene of New Jersey which represents the northernmost known occurrence of the species in eastern North America and the closest known occurrence of the species to a glacier or ice sheet in the continent during the Pleistocene. The specimen, which was collected at Ramanessin Brook in Holmdel, New Jersey, affirms that the Pleistocene deposits which line the banks of the popular Cretaceous site are not taphonomically biased to preserving larger fossils and in the future may yield an assemblage of small vertebrates.

scholarly journals The northernmost occurrence of Chelydra serpentina in the eastern US during the Pleistocene

2016 ◽  
Author(s):  
Chase Brownstein
Keyword(s):  
New Jersey ◽  
North America ◽  
Late Pleistocene ◽  
Ice Sheet ◽  
Eastern North America ◽  
Chelydra Serpentina ◽  
Turtle Species ◽  
Snapping Turtle ◽  
Caudal Vertebrae ◽  
Wide Range

The snapping turtle species Chelydra serpentina, which has a wide range across North America, is extremely tolerant to cold and even freezing conditions. Here, I describe a single caudal vertebrae referred to Chelydra serpentina from the Late Pleistocene of New Jersey which represents the northernmost known occurrence of the species in eastern North America and the closest known occurrence of the species to a glacier or ice sheet in the continent during the Pleistocene. The specimen, which was collected at Ramanessin Brook in Holmdel, New Jersey, affirms that the Pleistocene deposits which line the banks of the popular Cretaceous site are not taphonomically biased to preserving larger fossils and in the future may yield an assemblage of small vertebrates.

Darkening of Plant Tissues during in vitro Cultivation and Methods for its Prevention

2020 ◽  
Vol 36 (2) ◽  
pp. 26-42
Author(s):  
B.R. Kuluev
Keyword(s):  
Phenolic Compounds ◽  
Callus Formation ◽  
Plant Biotechnology ◽  
In Vitro Cultivation ◽  
Negative Effects ◽  
Wide Range ◽  
Negative Effect ◽  
Oxidative Transformations ◽  
Tissue Browning

One of the most common problems in the plant in vitro propagation is the tissue browning and subsequent necrosis, resulting from the oxidation of phenolic compounds, secondary metabolites produced in response to injury and released into the nutrient medium. This process is one of the main reasons for the decrease in the efficiency of callus formation, somatic embryogenesis, regeneration and genetic transformation of plants in vitro. Moreover, oxidative browning often leads to culture death. Therefore, the current problems in genetic and cellular engineering of a wide range of plant species can be solved only by preventing or reducing the negative effects of browning of in vitro cultures caused by the oxidative transformations of phenolic compounds into quinones toxic to cells. This review is devoted to the description of the main existing methods to prevent these adverse transformations. Various chemicals with antioxidant and adsorbing properties are used in plant biotechnology for this purpose, but there are no general approaches to solve the problem. Although the choice of the method to minimize the negative effect of phenolic compound oxidation depends, firs of all, on the species and variety of the plant, some agents, such as ascorbic acid, activated carbon, silver nitrate, can be considered as universal and quite effective in preventing oxidative darkening of explants in vitro. phenolic compounds, oxidative browning, polyphenol oxidase, tissue browning, in vitro, microclonal plant propagation The work was funded on the theme АААА-А17-117102740098-8.

scholarly journals Comparative study of fungal stability between Metarhizium strains after successive subculture

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Rana H. M. Hussien ◽  
Said M. Ezzat ◽  
Ali A. El Sheikh ◽  
James W. D. Taylor ◽  
Tariq M. Butt
Keyword(s):  
Growth Rate ◽  
Radial Growth ◽  
Morphological Characteristics ◽  
In Vitro Cultivation ◽  
Slight Variation ◽  
Main Body ◽  
Scale Production ◽  
Radial Growth Rate ◽  
Successive Subculture

Abstract Background Metarhizium species are considered one of the most outstanding powerful biological control agents that have been commercialized as biopesticides against various agricultural pests. Fungal stability with successive in vitro cultivation is a desirable trait for a large-scale production of fungal biopesticide. Main body The new Egyptian strain Metarhizium anisopliae AUMC 3262 exhibited auspicious results when compared to Metarhizium brunneum ARSEF 4556 and M. brunneum V275 based on the variations of fungal characteristics, and essential quality control parameters (radial growth rate, conidial yield, viability, and virulence) after repeated in vitro subculturing. Changes in morphological characteristics were noted at both AUMC 3262 and ARSEF 4556. Following the 5th subculture, decreased conidial yield was noted, though radial growth remained stable, confirming that there is a non-positive correlation between conidial yield and radial growth rate for these species. In contrast, V275 showed a high morphological stability, conidial yield, and radial growth rate after repeated subculture. The three tested strains manifested high viability up to 100% and displayed the same pattern of Pr1 production. A slight variation was recorded in the median lethal time (LT50) values against the great wax moth, Galleria mellonella (L.), larvae between different subcultures of the tested Metarhizium strains. Conclusion The new Egyptian strain AUMC 3262 showed a high stability with a slight difference in some parameters after the successive subculture compared to both ARSEF4556 and V275.

scholarly journals Zebrafish as an alternative method for toxicological studies

2020 ◽  
Author(s):  
Maria Sampieri ◽  
Riccardo Villa ◽  
Silvia Dotti
Keyword(s):  
Skin Sensitization ◽  
Industrial Chemicals ◽  
Tail Bud ◽  
In Vitro Methods ◽  
Regulatory Frameworks ◽  
Toxicological Studies ◽  
Somite Formation ◽  
Toxic Potential ◽  
3Rs Principle

According to the Directive 2010/63/EU fish embryos do not fall into regulatory frameworks dealing with animal experimentation. Therefore, in compliance with the 3Rs principle, zebrafish embryos are considered as replacement or refinement methods. Since more and more industrial chemicals are recognized causes of skin sensitization, it is needed a thorough understanding of the mechanisms to make predictions of the toxic potential of novel compounds. Thus, the FET test was performed and up to four apical observations were recorded as indicators of lethality: coagulation of fertilized eggs, lack of somite formation, no detachment of the tail bud from the yolk sac and lack of heartbeat. Then, in order to assess whether the skin sensitization due to chemical incubation was really measurable, the Fish Interleukin 8 (IL8) ELISA Kit was carried out. The preliminary results obtained so far seem encouraging. However, they need to be confirmed through further ELISA tests and compared with other in vitro methods.

scholarly journals Accumulation of Ascorbic Acid in Tomato Cell Culture: Influence of the Genotype, Source Explant and Time of In Vitro Cultivation

Antioxidants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 222 ◽  
Author(s):  
Maria Minutolo ◽  
Pasquale Chiaiese ◽  
Antonio Di Matteo ◽  
Angela Errico ◽  
Giandomenico Corrado
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Ascorbic Acid ◽  
Molecular Mechanisms ◽  
Cultured Cells ◽  
Biological Effects ◽  
Water Soluble ◽  
In Vitro Cultivation ◽  
Wide Range

The production and commercialization of natural antioxidants is gaining increasing importance due to their wide range of biological effects and applications. In vitro cell culture is a valuable source of plant bioactive compounds, especially those highly dependent on environmental factors. Nonetheless, research on the accumulation in plant cultured cells of water-soluble antioxidant vitamins, such as the ascorbic acid (AsA), is very limited. Tomato fruits are a main dietary source of vitamin C and in this work, we explored the potential of in vitro cultured cells for AsA accumulation. Specifically, using a full factorial design, we examined the effect of the source explant, the time in tissue culture and the genetic difference present in two Introgression Line (IL7-3 and IL12-4) that harbor Quantitative Trait Loci (QTLs) for ascorbic acid in fruits. Moreover, we performed an expression analysis of genes involved in AsA metabolism to highlight the molecular mechanisms that can account for the difference between fruit explants and calli. Our work indicated that cultured tomato cells accumulate AsA well beyond the amount present in fruits and that the three factors under investigation and their interaction significantly influence AsA accumulation. The time in tissue culture is the main single factor and, different from the expectations for secondary metabolites, explants from unripe, mature green fruits provided the highest increase in AsA. Moreover, in controlled conditions the genetic differences between the ILs and the control genotype are less relevant for calli cultivated for longer time. Our work showed the potential of tomato cell culture to produce AsA and prompt further refinements towards its possible large-scale exploitation.

Sign in / Sign up

Export Citation Format

Share Document