Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos

Development ◽  
1993 ◽  
Vol 119 (4) ◽  
pp. 1203-1215 ◽  
Author(s):  
C. Thisse ◽  
B. Thisse ◽  
T.F. Schilling ◽  
J.H. Postlethwait
Keyword(s):  
Neural Crest ◽  
Paraxial Mesoderm ◽  
Cloning And Expression ◽  
Tail Bud ◽  
No Tail ◽  
Mesoderm Formation ◽  
Snail Gene ◽  
Similar Gene ◽  
In Cells

Mesoderm formation is critical for the establishment of the animal body plan and in Drosophila requires the snail gene. This report concerns the cloning and expression pattern of the structurally similar gene snail1 from zebrafish. In situ hybridization shows that the quantity of snail1 RNA increases at the margin of the blastoderm in cells that involute during gastrulation. As gastrulation begins, snail1 RNA disappears from the dorsal axial mesoderm and becomes restricted to the paraxial mesoderm and the tail bud. snail1 RNA increases in cells that define the posterior border of each somite and then disappears when somitic cells differentiate. Later in development, expression appears in cephalic neural crest derivatives. Many snail1-expressing cells were missing from mutant spadetail embryos and the quantity of snail1 RNA was greatly reduced in mutant no tail embryos. The work presented here suggests that snail1 is involved in morphogenetic events during gastrulation, somitogenesis and development of the cephalic neural crest, and that no tail may act as a positive regulator of snail1.

scholarly journals Cloning and developmental expression of Sna, a murine homologue of the Drosophila snail gene

Development ◽  
1992 ◽  
Vol 116 (1) ◽  
pp. 227-237 ◽  
Author(s):  
M.A. Nieto ◽  
M.F. Bennett ◽  
M.G. Sargent ◽  
D.G. Wilkinson
Keyword(s):  
Neural Crest ◽  
Zinc Finger ◽  
In Situ Hybridisation ◽  
Developmental Expression ◽  
Dorsoventral Patterning ◽  
Murine Homologue ◽  
Extraembryonic Mesoderm ◽  
Snail Gene ◽  
Ectoplacental Cone

The genetic analysis of dorsoventral patterning in Drosophila has identified a zinc-finger gene, snail, that is required for mesoderm formation. The cloning and nuclease protection analysis of a Xenopus homologue of this gene has suggested a possible role in the mesoderm of vertebrates. Here, we describe the cloning of a murine homologue of snail, Sna, and in situ hybridisation studies of its developmental expression. Sequence analysis reveals substantial conservation of the second to fifth zinc fingers, but not of the first zinc finger in the Sna gene. Expression occurs in the ectoplacental cone, parietal endoderm, embryonic and extraembryonic mesoderm, in neural crest and in condensing precartilage. Based on the timing and spatial restriction of expression in embryonic mesoderm, we suggest that Sna might be required for the early development of this tissue, as is the case for its Drosophila counterpart. In addition, we propose that Sna might have an analogous role in the development of neural crest. The expression in condensing precartilage indicates that this gene also has a later function in chondrogenesis.

Detection of Nucleic Acids in Cells or Tissue Sections Using In situ Polymerase Chain Reaction (PCR)

1996 ◽  
Vol 54 ◽  
pp. 16-17
Author(s):  
J. R. Hully ◽  
K. R. Luehrsen ◽  
K. Aoyagi ◽  
C. Shoemaker ◽  
R. Abramson
Keyword(s):  
Nucleic Acids ◽  
Viral Infections ◽  
Anatomical Location ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Cellular Source ◽  
Gene Transcripts ◽  
Initial Description ◽  
In Cells

The development of PCR technology has greatly accelerated medical research at the genetic and molecular levels. Until recently, the inherent sensitivity of this technique has been limited to isolated preparations of nucleic acids which lack or at best have limited morphological information. With the obvious exception of cell lines, traditional PCR or reverse transcription-PCR (RT-PCR) cannot identify the cellular source of the amplified product. In contrast, in situ hybridization (ISH) by definition, defines the anatomical location of a gene and/or it’s product. However, this technique lacks the sensitivity of PCR and cannot routinely detect less than 10 to 20 copies per cell. Consequently, the localization of rare transcripts, latent viral infections, foreign or altered genes cannot be identified by this technique. In situ PCR or in situ RT-PCR is a combination of the two techniques, exploiting the sensitivity of PCR and the anatomical definition provided by ISH. Since it’s initial description considerable advances have been made in the application of in situ PCR, improvements in protocols, and the development of hardware dedicated to in situ PCR using conventional microscope slides. Our understanding of the importance of viral latency or viral burden in regards to HIV, HPV, and KSHV infections has benefited from this technique, enabling detection of single viral copies in cells or tissue otherwise thought to be normal. Clearly, this technique will be useful tool in pathobiology especially carcinogenesis, gene therapy and manipulations, the study of rare gene transcripts, and forensics.

Expression of a novel FGF in the Xenopus embryo. A new candidate inducing factor for mesoderm formation and anteroposterior specification

Development ◽  
1992 ◽  
Vol 114 (3) ◽  
pp. 711-720 ◽  
Author(s):  
H.V. Isaacs ◽  
D. Tannahill ◽  
J.M. Slack
Keyword(s):  
Specific Activity ◽  
Biological Properties ◽  
The Body ◽  
Mesoderm Induction ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Mesoderm Formation ◽  
Low Levels

We have cloned and sequenced a new member of the fibroblast growth factor family from Xenopus laevis embryo cDNA. It is most closely related to both mammalian kFGF (FGF-4) and FGF-6 but as it is not clear whether it is a true homologue of either of these genes we provisionally refer to it as XeFGF (Xenopus embryonic FGF). Two sequences were obtained, differing by 11% in derived amino acid sequence, which probably represent pseudotetraploid variants. Both the sequence and the behaviour of in vitro translated protein indicates that, unlike bFGF (FGF-2), XeFGF is a secreted molecule. Recombinant XeFGF protein has mesoderm-inducing activity with a specific activity similar to bFGF. XeFGF mRNA is expressed maternally and zygotically with a peak during the gastrula stage. Both probe protection and in situ hybridization showed that the zygotic expression is concentrated in the posterior of the body axis and later in the tailbud. Later domains of expression were found near the midbrain/hindbrain boundary and at low levels in the myotomes. Because of its biological properties and expression pattern, XeFGF is a good candidate for an inducing factor with possible roles both in mesoderm induction at the blastula stage and in the formation of the anteroposterior axis at the gastrula stage.

Onset of the segmentation clock in the chick embryo: evidence for oscillations in the somite precursors in the primitive streak

Development ◽  
2002 ◽  
Vol 129 (5) ◽  
pp. 1107-1117 ◽  
Author(s):  
Caroline Jouve ◽  
Tadahiro Iimura ◽  
Olivier Pourquie
Keyword(s):  
Continuous Process ◽  
Notch Pathway ◽  
Primitive Streak ◽  
The Body ◽  
Paraxial Mesoderm ◽  
Segmentation Clock ◽  
Head Mesoderm ◽  
Dynamic Expression ◽  
Mesoderm Formation ◽  
Cyclic Gene

Vertebrate somitogenesis is associated with a molecular oscillator, the segmentation clock, which is defined by the periodic expression of genes related to the Notch pathway such as hairy1 and hairy2 or lunatic fringe (referred to as the cyclic genes) in the presomitic mesoderm (PSM). Whereas earlier studies describing the periodic expression of these genes have essentially focussed on later stages of somitogenesis, we have analysed the onset of the dynamic expression of these genes during chick gastrulation until formation of the first somite. We observed that the onset of the dynamic expression of the cyclic genes in chick correlated with ingression of the paraxial mesoderm territory from the epiblast into the primitive streak. Production of the paraxial mesoderm from the primitive streak is a continuous process starting with head mesoderm formation, while the streak is still extending rostrally, followed by somitic mesoderm production when the streak begins its regression. We show that head mesoderm formation is associated with only two pulses of cyclic gene expression. Because such pulses are associated with segment production at the body level, it suggests the existence of, at most, two segments in the head mesoderm. This is in marked contrast to classical models of head segmentation that propose the existence of more than five segments. Furthermore, oscillations of the cyclic genes are seen in the rostral primitive streak, which contains stem cells from which the entire paraxial mesoderm originates. This indicates that the number of oscillations experienced by somitic cells is correlated with their position along the AP axis.

Overexpression of Snail family members highlights their ability to promote chick neural crest formation

Development ◽  
2002 ◽  
Vol 129 (7) ◽  
pp. 1583-1593 ◽  
Author(s):  
Marta G. del Barrio ◽  
M. Angela Nieto
Keyword(s):  
Chick Embryo ◽  
Neural Crest ◽  
Neural Tube ◽  
Family Members ◽  
Ectopic Expression ◽  
Endogenous Gene ◽  
The Family ◽  
Snail Family ◽  
Migratory Cells ◽  
Snail Gene

The Snail gene family of transcription factors plays crucial roles in different morphogenetic processes during the development of vertebrate and invertebrate embryos. In previous studies of function interference for one of the family members, Slug, we showed its involvement and neural crest formation in the chick embryo. Now we have carried out a series of gain-of-function experiments in which we show that Slug overexpression in the neural tube of the chick embryo induces an increase in neural crest production. The analysis of electroporated embryos shows that Slug can induce the expression of rhoB and an increase in the number of HNK-1-positive migratory cells, indicating that it lies upstream of them in the genetic cascade of neural crest development. The increase in neural crest production after Slug overexpression was confined to the cranial region, indicating that the mechanisms of crest induction somehow differ between head and trunk. The expression of the two vertebrate family members, Slug and Snail, is peculiar with respect to the neural crest. Slug is not expressed in the premigratory crest in the mouse, whereas it is expressed in this cell population in the chick and the opposite is true for Snail(Sefton, M., Sánchez, S. and Nieto M. A. (1998) Development125, 3111-3121). This raises the question of whether they can be functionally equivalent. To test this hypothesis both intra- and interspecies, we have performed a series of ectopic expression experiments by electroporating chick and mouse Snail in the chick embryo hindbrain. We observe that both genes elicit the same responses in the neural tube. Our results indicate that they can be functionally equivalent, although the embryos show a higher response to the endogenous gene, chick Slug.

scholarly journals Epha1 is a cell surface marker for neuromesodermal progenitors and their early mesoderm derivatives

2019 ◽  
Author(s):  
Luisa de Lemos ◽  
André Dias ◽  
Ana Nóvoa ◽  
Moisés Mallo
Keyword(s):  
Cell Surface ◽  
Transcriptional Profiling ◽  
Surface Marker ◽  
Cell Surface Marker ◽  
Paraxial Mesoderm ◽  
Molecular Signature ◽  
Tail Bud ◽  
Molecular Fingerprint ◽  
Simple Method ◽  
Cell Subpopulations

ABSTRACTThe vertebrate body is built during embryonic development by the sequential addition of new tissue as the embryo grows at its caudal end. During this process, the neuro-mesodermal progenitors (NMPs) generate the postcranial neural tube and paraxial mesoderm. Recently, several approaches have been designed to determine their molecular fingerprint but a simple method to isolate NMPs from embryos without the need for transgenic markers is still missing. We isolated NMPs using a genetic strategy that exploits their self-renew properties, and searched their transcriptome for cell surface markers. We found a distinct Epha1 expression profile in progenitor-containing areas of the mouse embryo, consisting of two cell subpopulations with different Epha1 expression levels. We show that Sox2+/T+ cells are preferentially associated with the Epha1 compartment, indicating that NMPs might be contained within this cell pool. Transcriptional profiling showed enrichment of high Epha1-expressing cells in known NMP and early mesoderm markers. Also, tail bud cells with lower Epha1 levels contained a molecular signature suggesting the presence of notochord progenitors. Our results thus indicate that Epha1 could represent a valuable cell surface marker for different subsets of axial progenitors, most particularly for NMPs taking mesodermal fates.

scholarly journals Dynamic expression of MMP28 during cranial morphogenesis

2020 ◽  
Vol 375 (1809) ◽  
pp. 20190559 ◽  
Author(s):  
Nadege Gouignard ◽  
Eric Theveneau ◽  
Jean-Pierre Saint-Jeannet
Keyword(s):  
Extracellular Matrix ◽  
Wound Healing ◽  
Neural Crest ◽  
Large Family ◽  
Tail Bud ◽  
Dynamic Expression ◽  
The Family ◽  
Extracellular Matrix Remodelling ◽  
And Migration ◽  
Human And Mouse

Matrix metalloproteinases (MMPs) are a large family of proteases comprising 24 members in vertebrates. They are well known for their extracellular matrix remodelling activity. MMP28 is the latest member of the family to be discovered. It is a secreted MMP involved in wound healing, immune system maturation, cell survival and migration. MMP28 is also expressed during embryogenesis in human and mouse. Here, we describe the detailed expression profile of MMP28 in Xenopus laevis embryos. We show that MMP28 is expressed maternally and accumulates at neurula and tail bud stages specifically in the cranial placode territories adjacent to migrating neural crest cells. As a secreted MMP, MMP28 may be required in neural crest–placode interactions. This article is part of a discussion meeting issue ‘Contemporary morphogenesis’.

Detection of transcription and translation in situ with biotinylated molecular probes in cells transfected with recombinant DNA plasmids

1986 ◽  
Vol 156 (1) ◽  
pp. 17-24 ◽  
Author(s):  
G.H. Smith ◽  
P.J. Doherty ◽  
R.B. Stead ◽  
C.M. Gorman ◽  
D.E. Graham ◽  
...  
Keyword(s):  
Recombinant Dna ◽  
Molecular Probes ◽  
In Cells

scholarly journals Expression of follistatin mRNA by somatotropes and mammotropes early in the rat estrous cycle.

1993 ◽  
Vol 41 (7) ◽  
pp. 955-960 ◽  
Author(s):  
B L Lee ◽  
G Unabia ◽  
G Childs
Keyword(s):  
Fetal Bovine Serum ◽  
S100 Protein ◽  
Thyroid Stimulating Hormone ◽  
Pituitary Cells ◽  
Folliculostellate Cells ◽  
Complementary Oligonucleotide ◽  
Fetal Bovine ◽  
In Cells ◽  
Stimulating Hormone

We previously found follistatin (FS) mRNA in gonadotropes [predominantly in cells with luteinizing hormone (LH) antigens] and folliculostellate cells (with S100 antigens) in diestrus rats pituitaries. However, earlier in the cycle, when percentages of gonadotropes are lowest, percentages of cells expressing FS are 1.5-2-fold higher than in diestrus. This study was designed to detect FS mRNA and other pituitary antigens to identify the additional cells with dual in situ hybridization and immunolabeling protocols. The mRNA was detected with biotinylated complementary oligonucleotide probes and avidin-biotin-peroxidase complexes. Significant labeling for FS mRNA was found in cells with the following antigens: growth hormone (GH) (7% of pituitary cells); prolactin (PRL) (5%); S100 protein (5%); follicle-stimulating hormone (FSH beta) (4%); LH beta (3%); and thyroid-stimulating hormone (TSH beta) (3%). Optimal conditions for detection included: overnight plating of > 50,000 cells/well (24-well tray) in media containing 10% fetal bovine serum; hybridization at 37 degrees C; and fixation in 2% glutaraldehyde. Whereas FS is expressed predominantly by LH gonadotropes at midcycle, FS mRNA can be expressed by all types of antigen-bearing cells earlier in the cycle. Its function in the pituitary may relate to its role in binding activin, which would result in inhibition of FSH release. However, since activin inhibits secretion of GH, PRL, and adrenocorticotropin (ACTH), FS may also control activin's effects on these cells. The FS-expressing cells may therefore be paracrine or autocrine regulators.

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