Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures

Yasui, George Shigueki; Bertolini, Rafaela Manchin; Suárez-López, Lucía; Xavier, Pedro Porfírio; Monzani, Paulo Sérgio; Nascimento, Nivaldo Ferreira do; Castilho, Antônio Leão; Nakaghi, Laura Satiko Okada; Santos, Silvio Carlos Alves dos; Senhorini, José Augusto

doi:10.1016/j.cryobiol.2020.06.004

Cited by 6 publications

(5 citation statements)

References 23 publications

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“…Triploids, tetraploids, gynogenetic, and chimeric fish were successfully developed by means of advanced biotechnologies ( Adamov et al, 2017 ; Nascimento et al, 2020a , 2020b ). In the Neotropical region, the yellowtail tetra became the most advanced with regards to such technologies, having important implications in embryology, genetics, reproduction, cryobiology, and even in medical sciences such as flow cytometry ( Santos et al, 2016 ; Xavier et al, 2017 ; Yasui et al, 2020a ). Several characin species present critical reproduction challenges or are considered endangered ( Silveira and Straube, 2008 ).…”

Section: Discussionmentioning

confidence: 99%

“…For instance, triploidization of Brycon amazonicus ( Nascimento et al, 2021 ) and Prochilodus lineatus ( Yasui et al, 2020b ) was successful when using the same procedures of heat shocking for second polar body retention (2 mpf for 2 min, 40°C). In addition, confirmation of the ploidy status using flow cytometry was achieved using the protocol established for yellowtail tetra ( Xavier et al, 2017 ; Yasui et al, 2020a ).…”

Section: Discussionmentioning

confidence: 99%

“…This technique is based on a two-step procedure with cell lysing and nuclear staining for the subsequent analysis. In addition, cold storage of the fin samples for the subsequent flow cytometric analysis was developed (Yasui et al, 2020a). Regarding chromosome Frontiers in Genetics frontiersin.org manipulation in a Neotropical species, the yellowtail tetra was the first to be analyzed using flow cytometry.…”

Section: Development Of Flow Cytometric Proceduresmentioning

confidence: 99%

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Establishing a model fish for the Neotropical region: The case of the yellowtail tetra Astyanax altiparanae in advanced biotechnology

et al. 2022

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The use of model organisms is important for basic and applied sciences. Several laboratory species of fishes are used to develop advanced technologies, such as the zebrafish (Danio rerio), the medaka (Oryzias latipes), and loach species (Misgurnus spp.). However, the application of these exotic species in the Neotropical region is limited due to differences in environmental conditions and phylogenetic distances. This situation emphasizes the establishment of a model organism specifically for the Neotropical region with the development of techniques that may be applicable to other Neotropical fish species. In this work, the previous research efforts are described in order to establish the yellowtail tetra Astyanax altiparanae as a model laboratory species for both laboratory and aquaculture purposes. Over the last decade, starting with artificial fertilization, the yellowtail tetra has become a laboratory organism for advanced biotechnology, such as germ cell transplantation, chromosome set manipulation, and other technologies, with applications in aquaculture and conservation of genetic resources. Nowadays, the yellowtail tetra is considered the most advanced fish with respect to fish biotechnology within the Neotropical region. The techniques developed for this species are being used in other related species, especially within the characins class.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Development Of Flow Cytometric Proceduresmentioning

confidence: 99%

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Establishing a model fish for the Neotropical region: The case of the yellowtail tetra Astyanax altiparanae in advanced biotechnology

et al. 2022

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“…Subsequently, the systems were solvated with the SPCE water model that specifies 3-center water molecules with appropriate charges. Further, Na + and Cl − ions were supplemented to the system at a concentration of 0.9%, tending to imitate natural piscine hemodynamic conditions [55][56][57][58] . The entire complex systems were subjected to energy minimization after the pre-processing stage employing the steepest descent algorithm for 5000 steps, each counting 2 fs.…”

Section: Molecular Dynamics Simulationsmentioning

confidence: 99%

Immunoinformatics approaches in developing a novel multi-epitope chimeric vaccine protective against Saprolegnia parasitica

Choudhury,

Kumar,

Nafidi

et al. 2024

Sci Rep

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Saprolegnia parasitica is responsible for devastating infections in fish and poses a tremendous threat to the global aquaculture industry. Presently, no safe and effective control measures are available, on the contrary, use of banned toxic compounds against the pathogen is affecting humans via biomagnification routes. This pioneering study aims to design an effective multi-epitope multi-target vaccine candidate against S. parasitica by targeting key proteins involved in the infection process. The proteins were analyzed and linear B-cell epitopes, MHC class I, and class II epitopes were predicted. Subsequently, highly antigenic epitopes were selected and fused to a highly immunogenic adjuvant, 50S ribosomal protein L7/L12, to design a multi-epitope chimeric vaccine construct. The structure of the vaccine was generated and validated for its stereochemical quality, physicochemical properties, antigenicity, allergenicity, and virulence traits. Molecular docking analyses demonstrated strong binding interactions between the vaccine and piscine immune receptors (TLR5, MHC I, MHC II). Molecular dynamics simulations and binding energy calculations of the complexes, further, reflected the stability and favorable interactions of the vaccine and predicted its cytosolic stability. Immune simulations predicted robust and consistent kinetics of the immune response elicited by the vaccine. The study posits the vaccine as a promising solution to combat saprolegniasis in the aquaculture industry.

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“…However, in the Neotropical region, species of importance for the economy such as Prochilodus lineatus (Reis et al 2016) or for basic and applied science such as Astyanax altiparanae (Yasui et al 2015(Yasui et al , 2022) have a more detailed biological description (De Souza et al 2015;Yasui et al 2020b). For example, A. altiparanae or yellow-tailed tetra, showed characteristics that facilitate its handling in the laboratory, such as a small size of 4-15 cm, remarkable fecundity (Yasui et al 2022) with spawning several times a year (Sato et al 2006)and omnivorous feeding habit that allow it to adapt to commercial feeding (Bertolini et al 2018), contributed to the yellow-tail tetra becoming an emerging model in studies such as chromosome manipulation (Adamov et al 2017; Nascimento et al 2020), cryopreservation (Yasui et al 2020a) among others (Rocha et al, 2023, Fernandes et al 2019.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the migratory pattern of Primordial Germ Cells in three neotropical fish species aiming genetic conservation actions

Rosero,

Monzani,

Pessoa

et al. 2023

Preprint

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Primordial germ cells (PGCs) are embryonic pluripotent cells that can differentiate into spermatogonia and oogonia, making them a genetic source for germplasm conservation through cryobanking and the generation of germline chimeras. Therefore, PGC studies should be conducted for each species, given possible divergences in migratory patterns of PGCs. In this work, the PGCs of 3 neotropical species (Pseudopimelodus mangurus, Astyanax altiparanae, and Prochilodus lineatus) were characterized by microinjection into zygotes of mRNA synthesized from the ddx4 3'UTR sequence of P. mangurus. Visualization of labeled PGCs was achieved by fluorescence microscopy during embryonic development. In addition, ddx4 and dnd1 expressions were evaluated during embryonic development, larvae, and adult tissues of P. mangurus, to validate their use as a PGC marker. As a result, the effective identification of PGCs was obtained. DsRed-positive PGC of P. mangurus was observed in the hatching stage, GFP-positive PGC of A. altiparanae in the gastrula stage, and GFP-positive PGCs from P. lineatus were identified at the segmentation stage, with representative labeling percentages of 29% and 16% in A. altiparanae and P. lineatus respectively. The expression of ddx4 and dnd1 of P. mangurus confirmed the specificity of these genes in germ cells. These results point to the functionality of the P. mangurus ddx4 3'UTR sequence as a PGC marker, demonstrating that PGC labeling was more efficient in A. altiparanae and P. lineatus. The procedures used to identify PGCs in P. mangurus consolidate the first step for generating germinal chimeras as a conservation action of P. mangurus.

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Flow cytometric analysis from fish samples stored at low, ultra-low and cryogenic temperatures

Cited by 6 publications

References 23 publications

Establishing a model fish for the Neotropical region: The case of the yellowtail tetra Astyanax altiparanae in advanced biotechnology

Establishing a model fish for the Neotropical region: The case of the yellowtail tetra Astyanax altiparanae in advanced biotechnology

Immunoinformatics approaches in developing a novel multi-epitope chimeric vaccine protective against Saprolegnia parasitica

Characterization of the migratory pattern of Primordial Germ Cells in three neotropical fish species aiming genetic conservation actions

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