Improved translation efficiency of injected mRNA during early embryonic development

Fink, Maria Cristina Domingues da Silva; Flekna, Gabriele; Ludwig, Alfred; Czerny, Thomas

doi:10.1002/dvdy.20995

Cited by 30 publications

(22 citation statements)

References 39 publications

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“…From our results, it is clear that long-lasting b-globin UTR mRNA was detected in zebrafish embryos compared to mRNA containing endogenous UTRs of the tol2 gene. The application of heterologous UTRs was selected as a strategy for injection experiments such as in a two-vector system, with pT7TS and pCS2 [31]. The pCS2 vector contains a CMV promoter, artificial UTRs, and the SV40 late poly(A) signal and was used for DNA injection experiments, which showed that artificial UTRs lack destabilizing elements and accordingly behave neutrally in embryos.…”

Section: Discussionmentioning

confidence: 99%

Using an improved Tol2 transposon system to produce transgenic zebrafish with epinecidin-1 which enhanced resistance to bacterial infection

Peng

Pan

Chou

et al. 2010

Fish & Shellfish Immunology

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Section: Discussionmentioning

confidence: 99%

Using an improved Tol2 transposon system to produce transgenic zebrafish with epinecidin-1 which enhanced resistance to bacterial infection

Peng

Pan

Chou

et al. 2010

Fish & Shellfish Immunology

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“…These restriction sites are required for insertion into the pMC expression vector, which caps the inserted fragment with the sequence elements of the ␤-globin 5Ј-UTR and 3Ј-UTR on both sides separately and SV40 poly(A) signal at the 3Ј tail additionally. This elevates the transcribed cRNA stability and activity about 100-fold in cRNA injection experiments (22). GFP cRNA was used as a control in cRNA rescue experiments, and it was prepared using pEGFP-C1 vector.…”

Section: Methodsmentioning

confidence: 99%

High Mobility Group Box-1 (HMGB1; Amphoterin) Is Required for Zebrafish Brain Development

Zhao

Kuja-Panula

Rouhiainen

et al. 2011

Journal of Biological Chemistry

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Hmgb1 (high mobility group box-1; amphoterin) is highly expressed in brain during early development of vertebrate and nonvertebrate species. However, its role in brain development remains elusive. Here we have cloned the zebrafish Hmgb1 and specifically manipulated Hmgb1 expression using injection of morpholino antisense oligonucleotides or Hmgb1 cRNA. The HMGB1 knockdown morphants produced by injection of three different morpholino oligonucleotides display a characteristic phenotype with smaller size, smaller brain width, and shorter distance between the eyes. Closer examination of the phenotype reveals severe defects in the development of the forebrain that largely lacks catecholaminergic neural networks. The HMGB1 morphant is deficient in survival and proliferation of neural progenitors and displays fewer cell groups expressing the transcription factor Pax6a in the forebrain and aberrant Wnt8 signaling. The mechanism of HMGB1-dependent progenitor survival involves the neuronal transmembrane protein AMIGO (amphoterin-induced gene and orf), the expression of which is regulated by HMGB1 in vivo. Our data demonstrate that HMGB1 is a critical factor for brain development, enabling survival and proliferation of neural progenitors that will form the forebrain structures.The high mobility group box-1 protein (HMGB1 3 ; also designated as HMG1 and amphoterin) is an abundantly occurring parental form of the HMG proteins (for review, see Ref. 1). HMGB1 is an exceptional member in the family of HMG-box proteins; depending on the cell type and its activation state, HMGB1 displays a nonnuclear localization and is secreted from cells, in contrast to most HMG-box proteins that are strictly bound to the cell nuclei (for recent reviews, see Refs. 2 and 3). During the last few years, the extensive literature dealing with HMGB1 functions has mainly focused on extracellular regulation of cells by HMGB1. HMGB1 can be passively released from injured cells, but it is also actively secreted due to several types of stimuli such as cell contact with extracellular matrix and cytokine stimulation of cells (2). Acetylation of lysine residues of HMGB1 has been shown to act as a signal leading to extracellular export via a non-classical secretory pathway (4). HMGB1 functions are currently mainly associated with binding to the cell surface receptor RAGE (receptor for advanced glycation end products), but Toll-like receptors have been increasingly suggested as membrane receptors of HMGB1 (for review, see Ref. 2).Compared with the extensive recent literature dealing with the pathophysiological roles of HMGB1 in inflammation, much less attention has been paid to its physiological roles. The Hmgb1 knock-out mouse survives until early postnatal age, and problems in glucose homeostasis have been suggested to cause multiorgan failure in these mice (5).HMGB1 was isolated from developing rat brain using neurite outgrowth in embryonic forebrain neurons as a readout in protein fractionation (6). These studies provided the initial evidence of HMGB1 as a...

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“…For transient reporter assays, the expression vector pMC (25) was used; for other cell culture experiments pKW10 (4) or pKC (polylinker modification of pKW) vectors (all three vectors contain cytomegalovirus promoters) were used. For all experiments, mouse Tle4, mouse Gbx2, and mouse Otx2 cDNAs were used.…”

Section: Methodsmentioning

confidence: 99%

Gbx2 and Otx2 Interact with the WD40 Domain of Groucho/Tle Corepressors

Heimbucher

Murko

Bajoghli

et al. 2007

Molecular and Cellular Biology

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One of the earliest organizational decisions in the development of the vertebrate brain is the division of the neural plate into Otx2-positive anterior and Gbx2-positive posterior territories. At the junction of these two expression domains, a local signaling center is formed, known as the midbrain-hindbrain boundary (MHB). This tissue coordinates or "organizes" the development of neighboring brain structures, such as the midbrain and cerebellum. Correct positioning of the MHB is thought to depend on mutual repression involving these two homeobox genes. Using a cell culture colocalization assay and coimmunoprecipitation experiments, we show that engrailed homology region 1 (eh1)-like motifs of both transcription factors physically interact with the WD40 domain of Groucho/Tle corepressor proteins. In addition, heat shock-induced expression of wild-type and mutant Otx2 and Gbx2 in medaka embryos demonstrates that Groucho is required for the repression of Otx2 by Gbx2. On the other hand, the repressive functions of Otx2 on Gbx2 do not appear to be dependent on corepressor interaction. Interestingly, the association of Groucho with Otx2 is also required for the repression of Fgf8 in the MHB. Therefore Groucho/Tle family members appear to regulate key aspects in the MHB development of the vertebrate brain.Gbx2 is a member of the homeobox gene family and has been identified in mammalian, avian, amphibian, and fish species (12,24,36,56,66). Gbx genes are related to the Drosophila unplugged gene, which is involved in tracheal branching (18). Gbx2 is a key player in the early patterning of the vertebrate brain and is expressed in the local signaling center known as the midbrain-hindbrain boundary (MHB) or isthmic organizer, which is positioned between the presumptive midbrain and hindbrain (reviewed in references 34, 54, 59, and 70). The Gbx2 expression domain is located at the region of the hindbrain, while the homeobox gene Otx2 is expressed in the presumptive forebrain and midbrain and thereby forms a common border with the Gbx2 domain at the position of the prospective MHB. Gbx2 mutant mice lack the anterior hindbrain and reveal a posterior expansion of the midbrain (67). In contrast, the anterior brain rostral to rhombomere 3 is deleted in Otx2-null mutant mice (3, 44). Misexpression of Gbx2 represses Otx2 expression in the posterior midbrain (46, 66), whereas misexpression of Otx2 in the anterior hindbrain represses Gbx2 expression in this region (11,35). Studies in Xenopus suggest that Otx and Gbx proteins needed for the positioning process function primarily as repressors rather than activators (28).Tle4 is one of the four full-length Groucho proteins in mammals (39, 63). The founding member of this conserved corepressor family is the Groucho gene of Drosophila. Groucho is known to play important roles in various developmental processes, including sex determination, segmentation, neurogenesis and dorsoventral patterning (22,50). Groucho family members are characterized by a conserved N-terminal glutamine-rich reg...

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Improved translation efficiency of injected mRNA during early embryonic development

Cited by 30 publications

References 39 publications

Using an improved Tol2 transposon system to produce transgenic zebrafish with epinecidin-1 which enhanced resistance to bacterial infection

Using an improved Tol2 transposon system to produce transgenic zebrafish with epinecidin-1 which enhanced resistance to bacterial infection

High Mobility Group Box-1 (HMGB1; Amphoterin) Is Required for Zebrafish Brain Development

Gbx2 and Otx2 Interact with the WD40 Domain of Groucho/Tle Corepressors

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