Isolation and characterization of a downstream target of Pax6 in the mammalian retinal primordium

Development ◽  
2001 ◽  
Vol 128 (20) ◽  
pp. 3987-3994 ◽  
Author(s):  
Gilbert Bernier ◽  
Wolfgang Vukovich ◽  
Lorenz Neidhardt ◽  
Bernhard G. Herrmann ◽  
Peter Gruss
Keyword(s):  
Transcription Factor ◽  
Expression Pattern ◽  
Large Scale ◽  
Ectopic Expression ◽  
Signal Transduction Pathway ◽  
Homeobox Gene ◽  
Optic Vesicle ◽  
Altered Expression ◽  
Retina Development ◽  
Isolation And Characterization

The transcription factor Pax6 is required for eye morphogenesis in humans, mice and insects, and can induce ectopic eye formation in vertebrate and invertebrate organisms. Although the role of Pax6 has intensively been studied, only a limited number of genes have been identified that depend on Pax6 activity for their expression in the mammalian visual system. Using a large-scale in situ hybridization screen approach, we have identified a novel gene expressed in the mouse optic vesicle. This gene, Necab, encodes a putative cytoplasmic Ca2+-binding protein and coincides with Pax6 expression pattern in the neural ectoderm of the optic vesicle and in the forebrain pretectum. Remarkably, Necab expression is absent in both structures in Pax6 mutant embryos. By contrast, the optic vesicle-expressed homeobox genes Rx, Six3, Otx2 and Lhx2 do not exhibit an altered expression pattern. Using gain-of-function experiments, we show that Pax6 can induce ectopic expression of Necab, suggesting that Necab is a direct or indirect transcriptional target of Pax6. In addition, we have found that Necab misexpression can induce ectopic expression of the homeobox gene Chx10, a transcription factor implicated in retina development. Taken together, our results provide evidence that Necab is genetically downstream of Pax6 and that it is a part of a signal transduction pathway in retina development.

Identifying targets of the rough homeobox gene of Drosophila: evidence that rhomboid functions in eye development

Development ◽  
1992 ◽  
Vol 116 (2) ◽  
pp. 335-346 ◽  
Author(s):  
M. Freeman ◽  
B.E. Kimmel ◽  
G.M. Rubin
Keyword(s):  
Cell Fate ◽  
Target Genes ◽  
Eye Development ◽  
Expression Patterns ◽  
Ectopic Expression ◽  
Homeobox Gene ◽  
Homeodomain Protein ◽  
Dorsoventral Patterning ◽  
Altered Expression ◽  
Enhancer Trap Lines

In order to identify potential target genes of the rough homeodomain protein, which is known to specify some aspects of the R2/R5 photoreceptor subtype in the Drosophila eye, we have carried out a search for enhancer trap lines whose expression is rough-dependent. We crossed 101 enhancer traps that are expressed in the developing eye into a rough mutant background, and have identified seven lines that have altered expression patterns. One of these putative rough target genes is rhomboid, a gene known to be required for dorsoventral patterning and development of some of the nervous system in the embryo. We have examined the role of rhomboid in eye development and find that, while mutant clones have only a subtle phenotype, ectopic expression of the gene causes the non-neuronal mystery cells to be transformed into photoreceptors. We propose that rhomboid is a part of a partially redundant network of genes that specify photoreceptor cell fate.

Ectopic expression of the maize kn1 gene phenocopies the Hooded mutant of barley

Development ◽  
1997 ◽  
Vol 124 (19) ◽  
pp. 3737-3745 ◽  
Author(s):  
R.E. Williams-Carrier ◽  
Y.S. Lie ◽  
S. Hake ◽  
P.G. Lemaux
Keyword(s):  
Expression Pattern ◽  
Posttranscriptional Regulation ◽  
Cellular Differentiation ◽  
Ectopic Expression ◽  
Homeobox Gene ◽  
Mrna Localization ◽  
Constitutive Promoter ◽  
Homeotic Transformation ◽  
Shoot Meristems

The homeobox gene, knotted1, (kn1) is expressed in shoot meristems and is required for maintaining indeterminacy and preventing cellular differentiation. Awns, extensions of the bract-like lemma found in all grass inflorescences, are normally determinate structures. We show that ectopic expression of kn1 in the barley awn is sufficient to direct the development of ectopic meristems, forming inflorescence-like structures. This homeotic transformation is similar to the phenotype produced by misexpression of the barley hvknox3 gene, associated with the dominant Hooded mutant (Muller, K. J., Romano, N., Gerstner, O., Garcia-Maroto, F., Pozzi, C., Salamini, F. and Rohde, W. (1995) Nature 374, 727–730). We suggest that the inverse polarity of the ectopic flowers seen in Hooded and transgenic kn1 plants results from the transformation of the awn into reiterative inflorescence axes. We observed that the protein and mRNA localization of the transgene, driven by a constitutive promoter, is similar to the expression pattern of hvknox3 in awns of Hooded mutants, suggesting posttranscriptional regulation.

scholarly journals Arabidopsis LSH8 Positively Regulates ABA Signaling by Changing the Expression Pattern of ABA-Responsive Proteins

2021 ◽  
Vol 22 (19) ◽  
pp. 10314
Author(s):  
Jinpeng Zou ◽  
Zhifang Li ◽  
Haohao Tang ◽  
Li Zhang ◽  
Jingdu Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Seed Germination ◽  
Signaling Pathway ◽  
Expression Pattern ◽  
Large Scale ◽  
Aba Signaling ◽  
Wild Type ◽  
Floral Organogenesis ◽  
Early Seedling ◽  
Aba Insensitive

Phytohormone ABA regulates the expression of numerous genes to significantly affect seed dormancy, seed germination and early seedling responses to biotic and abiotic stresses. However, the function of many ABA-responsive genes remains largely unknown. In order to improve the ABA-related signaling network, we conducted a large-scale ABA phenotype screening. LSH, an important transcription factor family, extensively participates in seedling development and floral organogenesis in plants, but whether its family genes are involved in the ABA signaling pathway has not been reported. Here we describe a new function of the transcription factor LSH8 in an ABA signaling pathway. In this study, we found that LSH8 was localized in the nucleus, and the expression level of LSH8 was significantly induced by exogenous ABA at the transcription level and protein level. Meanwhile, seed germination and root length measurements revealed that lsh8 mutant lines were ABA insensitive, whereas LSH8 overexpression lines showed an ABA-hypersensitive phenotype. With further TMT labeling quantitative proteomic analysis, we found that under ABA treatment, ABA-responsive proteins (ARPs) in the lsh8 mutant presented different changing patterns with those in wild-type Col4. Additionally, the number of ARPs contained in the lsh8 mutant was 397, six times the number in wild-type Col4. In addition, qPCR analysis found that under ABA treatment, LSH8 positively mediated the expression of downstream ABA-related genes of ABI3, ABI5, RD29B and RAB18. These results indicate that in Arabidopsis, LSH8 is a novel ABA regulator that could specifically change the expression pattern of APRs to positively mediate ABA responses.

Expression of Fetal Hemoglobin among Adult Human Erythroblasts Is Associated with an Altered Expression Pattern for the Transcription Factor c-Myb.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3635-3635
Author(s):  
Parvathi Rudrabhatla ◽  
Jeffery L. Miller
Keyword(s):  
Transcription Factor ◽  
Expression Pattern ◽  
Quantitative Pcr ◽  
Terminal Differentiation ◽  
Fetal Hemoglobin ◽  
Major Band ◽  
Altered Expression ◽  
Adult Human ◽  
Nuclear Extracts ◽  
Factor C

Abstract The c-Myb transcription factor plays a central role in regulating growth and differentiation during definitive erythropoiesis. In addition, c-Myb expression has been previously linked to GATA-1 expression and hereditary persistence of fetal hemoglobin. Here we examined a potential role for c-Myb in signaled expression of fetal hemoglobin (HbF) among primary adult human erythroblasts. Initial experiments were performed to determine the expression pattern of c-Myb under conditions of low fetal hemoglobin production (CD34 cells cultured over 14 days in media supplemented with erythropoietin (EPO) alone). Quantitative PCR performed on cells sampled on days 2, 5, 8, 11, and 14 demonstrated highly regulated c-Myb expression. A dramatic increase (>30 fold) in c-Myb mRNA was measured between days 5 and 8 as the committed progenitor cells differentiated to the proerythroblast stage. That increase was followed by a 3-fold reduction in c-Myb during terminal differentiation. Western blotting of nuclear extracts obtained on days 7 and 14 in EPO cultures corroborated with the quantitative PCR analyses. Electrophoretic mobility shift assays performed using day 7 and day 14 nuclear extracts showed a shifted major band with a c-Myb consensus probe. That major band was competed by unlabeled c-Myb probe on both days. Moreover, day 14 extracts repeatedly revealed two additional bands that were not competed suggesting the possibility of other complexes binding to the Myb motif or changes in c-Myb conformation during terminal differentiation. Chromatin immunoprecipitation assays also revealed preliminary evidence for Myb binding directly to beta globin locus. When compared to low HbF culture conditions (EPO; HbF <1%), cells grown in high HbF conditions (erythropoietin + stem cell factor + TGF-beta (EST); HbF>35%) demonstrated a distinct pattern of c-Myb expression. Relative to the low HbF conditions, high HbF expressing cells had reduced c-Myb levels on day 7 followed by increased levels on day 14. These results suggest that c-Myb possesses highly regulated patterns of expression and DNA binding during erythroid maturation. In addition, the association of changes in c-Myb expression patterns with HbF production suggests that this growth-related transcription factor may help regulate fetal hemoglobin synthesis in patients with beta-hemoglobinopathies.

Hhex and scl function in parallel to regulate early endothelial and blood differentiation in zebrafish

Development ◽  
2000 ◽  
Vol 127 (20) ◽  
pp. 4303-4313 ◽  
Author(s):  
W. Liao ◽  
C.Y. Ho ◽  
Y.L. Yan ◽  
J. Postlethwait ◽  
D.Y. Stainier
Keyword(s):  
Transcription Factor ◽  
Blood Cells ◽  
Early Stage ◽  
Ectopic Expression ◽  
Homeobox Gene ◽  
Transcription Factor Gene ◽  
Factor Gene ◽  
Molecular Events ◽  
Initial Framework

During embryogenesis, endothelial and blood precursors are hypothesized to arise from a common progenitor, the hemangioblast. Several genes that affect the differentiation of, or are expressed early in, both the endothelial and blood lineages may in fact function at the level of the hemangioblast. For example, the zebrafish cloche mutation disrupts the differentiation of both endothelial and blood cells. The transcription factor gene scl is expressed in both endothelial and blood lineages from an early stage and can regulate their differentiation. Here we report that in zebrafish the homeobox gene hhex (previously called hex) is also expressed in endothelial and blood lineages from an early stage. We find that hhex expression in these lineages is significantly reduced in cloche mutant embryos, indicating that hhex functions downstream of cloche to regulate endothelial and blood differentiation. Ectopic expression of hhex through injection of a DNA construct leads to the premature and ectopic expression of early endothelial and blood differentiation genes such as fli1, flk1 and gata1, indicating that Hhex can positively regulate endothelial and blood differentiation. However, analysis of a hhex deficiency allele shows that hhex is not essential for early endothelial and blood differentiation, suggesting that another gene, perhaps scl, compensates for the absence of Hhex function. Furthermore, we find that hhex and scl can induce each other's expression, suggesting that these two genes cross-regulate each other during early endothelial and blood differentiation. Together, these data provide the initial framework of a pathway that can be used to further integrate the molecular events regulating hemangioblast differentiation.

scholarly journals Analysis of maxillopedia Expression Pattern and Larval Cuticular Phenotype in Wild-Type and Mutant Tribolium

Genetics ◽  
2000 ◽  
Vol 155 (2) ◽  
pp. 721-731 ◽  
Author(s):  
Teresa D Shippy ◽  
Jianhua Guo ◽  
Susan J Brown ◽  
Richard W Beeman ◽  
Robin E Denell
Keyword(s):  
Rna Interference ◽  
Expression Pattern ◽  
Tribolium Castaneum ◽  
Expression Patterns ◽  
Ectopic Expression ◽  
Homeotic Gene ◽  
Wild Type ◽  
Double Stranded Rna ◽  
Similar Expression ◽  
Mutant Phenotypes

Abstract The Tribolium castaneum homeotic gene maxillopedia (mxp) is the ortholog of Drosophila proboscipedia (pb). Here we describe and classify available mxp alleles. Larvae lacking all mxp function die soon after hatching, exhibiting strong transformations of maxillary and labial palps to legs. Hypomorphic mxp alleles produce less severe transformations to leg. RNA interference with maxillopedia double-stranded RNA results in phenocopies of mxp mutant phenotypes ranging from partial to complete transformations. A number of gain-of-function (GOF) mxp alleles have been isolated based on transformations of adult antennae and/or legs toward palps. Finally, we have characterized the mxp expression pattern in wild-type and mutant embryos. In normal embryos, mxp is expressed in the maxillary and labial segments, whereas ectopic expression is observed in some GOF variants. Although mxp and Pb display very similar expression patterns, pb null embryos develop normally. The mxp mutant larval phenotype in Tribolium is consistent with the hypothesis that an ancestral pb-like gene had an embryonic function that was lost in the lineage leading to Drosophila.

scholarly journals Complexity of Developmental Control: Analysis of Embryonic Cell Lineage Specification in Caenorhabditis elegans Using pes-1 as an Early Marker

Genetics ◽  
1999 ◽  
Vol 151 (1) ◽  
pp. 131-141
Author(s):  
Laurent Molin ◽  
Heinke Schnabel ◽  
Titus Kaletta ◽  
Richard Feichtinger ◽  
Ian A Hope ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Expression Pattern ◽  
Fusion Gene ◽  
Ectopic Expression ◽  
Gene Expression Pattern ◽  
Embryonic Cell ◽  
Control Analysis ◽  
Normal Pattern ◽  
Variable Expression ◽  
Founder Cells

Abstract In the early Caenorhabditis elegans embryo five somatic founder cells are born during the first cleavages. The first of these founder cells, named AB, gives rise to 389 of the 558 nuclei present in the hatching larva. Very few genes directly involved in the specification of the AB lineage have been identified so far. Here we describe a screen of a large collection of maternal-effect embryonic lethal mutations for their effect on the early expression of a pes-1::lacZ fusion gene. This fusion gene is expressed in a characteristic pattern in 14 of the 32 AB descendants present shortly after the initiation of gastrulation. Of the 37 mutations in 36 genes suspected to be required specifically during development, 12 alter the expression of the pes-1::lacZ marker construct. The gene expression pattern alterations are of four types: reduction of expression, variable expression, ectopic expression in addition to the normal pattern, and reduction of the normal pattern together with ectopic expression. We estimate that ∼100 maternal functions are required to establish the pes-1 expression pattern in the early embryo.

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