Multiple Levels of Posttranscriptional Control Lead to Germ Line-Specific Gene Expression in the Zebrafish

Wolke, Uta; Weidinger, Gilbert; Köprunner, Marion; Raz, Erez

doi:10.1016/s0960-9822(02)00679-6

Cited by 120 publications

(115 citation statements)

References 19 publications

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“…This high level persists until the early gastrula stage and declines dramatically to a low level afterward. This temporal expression pattern is reminiscent of the zebrafish situation, where vasa RNA is also high until the early gastrula stage at 6 hr postfertilization and decreases sharply to a low level (Wolke et al, 2002). In zebrafish, vasa RNA is a component of the germ plasm and segregates with PGCs (Knaut et al, 2000(Knaut et al, ,2002.…”

Section: Discussionmentioning

confidence: 94%

“…Furthermore, CagVasa shares with Vasa proteins an important common feature, namely, the presence of multiple N-terminal RGG repeats that are thought to be a putative RNA-binding motif (Liang et al, 1994). Recently, the RGG-rich N-terminus of the zebrafish Vasa protein has been implicated in subcellular localization of the fused green fluorescent protein in PGCs (Wolke et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

“…Like Vasa homologs, CagVasa is characterized by an RGGrich N-terminus and eight diagnostic motifs: AQTGSGKT as the ATPase motif A; PTRELI, GG, TPGR, and DEAD as the ATPase motif B; SAT and ARGXD for RNA unwinding; and HRIGRTGR for RNA binding. The RGG repeats at the N-terminus possibly functions in RNA binding (Hay et al, 1988;Liang et al, 1994) and in subcellular protein localization (Wolke et al, 2002). These eight motifs are highly conserved between CagVasa and representative Vasa proteins from invertebrates and vertebrates on a sequence alignment (Fig.…”

Section: Isolation Of a Full-length Vasa Cdnamentioning

confidence: 99%

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Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate

Gui

Hong

2005

Developmental Dynamics

151

132

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The RNA helicase Vasa is a germ cell marker in animals, and its homolog in vertebrates to date has been limited to bisexual reproduction. We cloned and characterized CagVasa, a Vasa homolog from the gibel carp, a fish that reproduces bisexually or gynogenetically. CagVasa possesses 14 RGG repeats and eight conserved motifs of Vasa proteins. In bisexually reproducing gibel carp, vasa is maternally supplied and its zygotic expression is restricted to gonads. By in situ hybridization on testicular sections, vasa is low in spermatogonia, high in primary spermatocytes, reduced in secondary spermatocytes, but disappears in spermatids and sperm. In contrast, vasa persists throughout oogenesis, displaying low-high-low levels from oogonia over vitellogenic oocytes to maturing oocytes. A rabbit anti-Vasa antibody (␣Vasa) was raised against the N-terminal CagVasa for fluorescent immunohistochemistry. On testicular sections, Vasa is the highest in spermatogonia, reduced in spermatocytes, low in spermatids, and absent in sperm. In the ovary, Vasa is the highest in oogonia but persists throughout oogenesis. Subcellular localization of vasa and its protein changes dynamically during oogenesis. The ␣Vasa stains putative primordial germ cells in gibel carp fry. It detects gonadal germ cells also in several other teleosts. Therefore, Cagvasa encodes a Vasa ortholog that is differentially expressed in the testis and ovary. Interestingly, the ␣Vasa in combination with a nuclear dye can differentiate critical stages of spermatogenesis and oogenesis in fish. The cross-reactivity and the ability to stain stage-specific germ cells make this antibody a useful tool to identify fish germ cell development and differentiation. Developmental Dynamics 233:872-882, 2005.

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Section: Isolation Of a Full-length Vasa Cdnamentioning

confidence: 99%

See 1 more Smart Citation

Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate

Gui

Hong

2005

Developmental Dynamics

151

132

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“…With the occurrence of a novel technique, localized RNA expression (LRE), the germ cell marker gene, vasa, was utilized to label living PGCs (Wolke et al 2002). Briefly, synthetic chimeric mRNA constructed by fusing the green/red fluorescent protein (GFP/RFP) coding region to vasa 3'-untranslated region (3'UTR) can be used to visualize PGCs during embryogenesis, following injection into fertilized eggs.…”

Section: Electronic Supplementary Materialsmentioning

confidence: 99%

Identification and sexually dimorphic expression of vasa isoforms in Dabry′s sturgeon (Acipenser dabryanus), and functional analysis of vasa 3′-untranslated region

Yue

Yang

et al. 2016

Cell Tissue Res

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Germ cells are set aside from somatic cells early in embryogenesis, and are responsible for transmitting genetic information through generations. Vasa is a highly conserved germ cell marker across animal phyla, and widely used to label primordial germ cells. Dabry's sturgeon is a rare and endangered species distributed solely in the Yangtze River basin. Here, seven vasa isoforms, named Advasa1-7, were isolated and characterized in Dabry's sturgeon. RT-PCR and western blot analyses revealed that vasa mRNA and protein were mainly restricted to the testis and ovary, but exhibited sexually dimorphic expression. Cellular and subcellular localization uncovered that Advasa mRNA and protein displayed mitotic and meiotic expression in females, and mainly showed mitotic expression in males; surprisingly, they exhibited both cytoplasmic and nuclear expression in the ovarian germ cells, while showing exclusively cytoplasmic expression in the testicular germ cells. By microinjecting chimeric RNA consisting of the red fluorescent protein coding region and the Advasa 3'-untranslated region into embryos of Dabry's sturgeon, zebrafish and medaka, we demonstrated that it had the ability to visualize primordial germ cells (PGCs) in Dabry's sturgeon and zebrafish but not in medaka. It seemed that the machinery of vasa 3'UTR RNA localization was conserved between Dabry's sturgeon and ostariophysan, while possibly changed during the divergence of euteleosts and ostariophysan. Finally, Dabry's sturgeon PGCs moved on the yolk ball, and migrated toward the genital ridge via mesenchyme. Taken together, these results provide new information for vasa expression pattern and function, and lay a foundation for PGC cryopreservation and conservation of Dabry's sturgeon.

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“…Employing the kop promoter to establish a transgenic fish line expressing EGFP-F during early PGC development To follow PGC migration in live embryos, we have routinely labelled these cells with GFP relying on 3ЈUTR elements that confer specific expression in PGCs (Köprunner et al, 2001;Wolke et al, 2002). Others have used transgenic fish in which GFP was flanked by an upstream EF1␣ or vasa promoter and by the downstream 3ЈUTR of the vasa gene Krøvel and Olsen, 2002).…”

Section: Alterations In the Level Of Kop Activitymentioning

confidence: 99%

Transition from non-motile behaviour to directed migration during early PGC development in zebrafish

Blaser

Eisenbeiß

Neumann

et al. 2005

Journal of Cell Science

Self Cite

165

189

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The migration of zebrafish primordial germ cells (PGCs) is directed by SDF-1a and serves as a model for long-range chemokine-guided cell migration. Whereas the development and migration of zebrafish PGCs have been studied in great detail starting at mid-gastrulation stages when the cells exhibit guided active migration [7-8 hours post fertilization (hpf)], earlier stages have not yet been examined. Here we show that the PGCs acquire competence to respond to the chemokine following discrete maturation steps. Using the promoter of the novel gene askopos and RNA elements of nanos1 to drive GFP expression in PGCs, we found that immediately after their specification (about 3 hpf) PGCs exhibit simple cell shape. This stage is followed by a phase at which the cells assume complex morphology yet they neither change their position nor do they respond to SDF-1a. During the third phase, a transition into a `migratory stage' occurs as PGCs become responsive to directional cues provided by somatic cells secreting the chemokine SDF-1a. This transition depends on zygotic transcription and on the function of the RNA-binding protein Dead end and is correlated with down regulation of the cell adhesion molecule E-cadherin. These distinctive morphological and molecular alterations could represent a general occurrence in similar processes critical for development and disease.

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Multiple Levels of Posttranscriptional Control Lead to Germ Line-Specific Gene Expression in the Zebrafish

Cited by 120 publications

References 19 publications

Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate

Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp (Carassius auratus gibelio), a bisexually and gynogenetically reproducing vertebrate

Identification and sexually dimorphic expression of vasa isoforms in Dabry′s sturgeon (Acipenser dabryanus), and functional analysis of vasa 3′-untranslated region

Transition from non-motile behaviour to directed migration during early PGC development in zebrafish

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