Characterization of vasa and dnd homologs in summer flounder, Paralichthys dentatus: Expression analysis and colocalization of PGCs during embryogenesis

Du, Shuran; Zhou, Li; Xu, Shihong; Li, Jun; Song, Zongcheng; Liu, Qinghua

doi:10.1016/j.theriogenology.2022.01.007

Cited by 8 publications

(4 citation statements)

References 48 publications

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“…Moreover, dual color FISH is used to compare precisely the RNA expression patterns between Cldnd and Clvasa . The results of co-localization show that there is almost no difference between Cldnd and Clvasa in the testis, which is in line with the expression pattern in summer flounder ( Paralichthys dentatus ) [ 49 ]. In the ovary, Cldnd RNA is always uniformly distributed in the cytoplasm of germ cells, which differs from the specific expression of Clvasa in oocytes of stage I.…”

Section: Discussionsupporting

confidence: 66%

Spinyhead Croaker Germ Cells Gene dnd Visualizes Primordial Germ Cells in Medaka

Liu

Wen

et al. 2022

Life

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Spinyhead croaker (Collichthys lucidus) is an economically important fish suffering from population decline caused by overfishing and habitat destruction. Researches on the development of primordial germ cell (PGC) and reproduction biology were an emergency for the long-term conservation of the involved species. Dead end (dnd) gene plays an indispensable role in PGC specification, maintenance, and development. In the current study, we report the cloning and expression patterns of dnd in C. lucidus (Cldnd). RT-PCR analysis revealed that Cldnd was specifically expressed in both sexual gonads. In the ovary, Cldnd RNA was uniformly distributed in the oocytes and abundant in oogonia, and gradually decreased with oogenesis. A similar expression pattern was also detected in testis. Dual fluorescent in situ hybridization of Cldnd and Clvasa demonstrated that they almost had the same distribution except in oocytes at stage I, in which the vasa RNA aggregated into some particles. Furthermore, Cldnd 3′ UTR was sufficient to guide the Green Fluorescent Protein (GFP) specifically and stably expressed in the PGCs of medaka. These findings offer insight into that Cldnd is an evolutionarily conserved germline-specific gene and even a potential candidate for PGC manipulation in C. lucidus.

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Section: Discussionsupporting

confidence: 66%

Spinyhead Croaker Germ Cells Gene dnd Visualizes Primordial Germ Cells in Medaka

Liu

Wen

et al. 2022

Life

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“…The semi-quantitative PCR results in S. salar revealed that the vasa transcript was expressed up to the 10-somite stage, and dnd1 was expressed up to the early blastula stage, which corresponds to 162 d°C and 30 d°C [at 6 °C], respectively [ 79 ]. In summer flounder ( Paralichthys dentatus ) and Nile tilapia ( Oreochromis niloticus ), both the transcripts were expressed up to the blastula stage [ 30 , 80 ]. The mRNA expression pattern of the two transcripts suggests that they are maternally inherited [ 79 , 81 ] and play an important role in the PGC specification and development.…”

Section: Discussionmentioning

confidence: 99%

Insights into Early Ontogenesis of Salmo salar: RNA Extraction, Housekeeping Gene Validation and Transcriptional Expression of Important Primordial Germ Cell and Sex-Determination Genes

Bhat

Dubiel

Rodrı́guez³

et al. 2023

Animals

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The challenge in extracting high-quality RNA impedes the investigation of the transcriptome of developing salmonid embryos. Furthermore, the mRNA expression pattern of important PGC and SD genes during the initial embryonic development of Salmo salar is yet to be studied. So, in the present study, we aimed to isolate high-quality RNA from eggs and developing embryos to check vasa, dnd1, nanos3a, sdf1, gsdf, amh, cyp19a, dmrt1 and foxl2 expression by qPCR. Additionally, four HKGs (GAPDH, UB2L3, eEf1a and β-actin) were validated to select the best internal control for qPCR. High-quality RNA was extracted, which was confirmed by spectrophotometer, agarose gel electrophoresis and Agilent TapeStation analysis. UB2L3 was chosen as a reference gene because it exhibited lower intra- and inter-sample variation. vasa transcripts were expressed in all the developmental stages, while dnd1 was expressed only up to 40 d°C. Nanos3a was expressed in later stages and remained at its peak for a shorter period, while sdf1 showed an irregular pattern of mRNA expression. The mRNA expression levels of SD genes were observed to be upregulated during the later stages of development, prior to hatching. This study presents a straightforward methodology for isolating high-quality RNA from salmon eggs, and the resulting transcript profiles of significant PGC and SD genes in S. salar could aid in improving our comprehension of reproductive development in this commercially important species.

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“…The expressions of ddx4 and dnd1 of P. mangurus were not identi ed in the gastrula and somites stages, contrary to other patterns observed in several teleost species; for example, Salmo salar showed a progressive decrease of dnd1 and a constant expression of ddx4 up to 10 somites (Nagasawa et al 2013), T. macdonaldi showed progressive decrease of ddx4 until hatching in(Escuredo-Vielba et al 2018) similar to R. quelen (Ricci et al 2018) and in Paralichthys dentatus the expression level of dnd1 and ddx4 increased until it reached a peak during cleavage at the 16-32 cell stage with a progressive decrease during gastrulation and segmentation (Du et al 2022), demonstrating the differences in gene expression patterns between the species during embryonic development. In female and male tissues, ddx4 expression was observed in the gonads, corroborating that the gene's functions and expression are maintained throughout germ cell development and in post-meiotic spermatids and oocytes (Gassei et al 2017).…”

Section: Discussionmentioning

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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Characterization of vasa and dnd homologs in summer flounder, Paralichthys dentatus: Expression analysis and colocalization of PGCs during embryogenesis

Cited by 8 publications

References 48 publications

Spinyhead Croaker Germ Cells Gene dnd Visualizes Primordial Germ Cells in Medaka

Spinyhead Croaker Germ Cells Gene dnd Visualizes Primordial Germ Cells in Medaka

Insights into Early Ontogenesis of Salmo salar: RNA Extraction, Housekeeping Gene Validation and Transcriptional Expression of Important Primordial Germ Cell and Sex-Determination Genes

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

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