Differential induction of four msx homeobox genes during fin development and regeneration in zebrafish

Development ◽  
1995 ◽  
Vol 121 (2) ◽  
pp. 347-357 ◽  
Author(s):  
M.A. Akimenko ◽  
S.L. Johnson ◽  
M. Westerfield ◽  
M. Ekker
Keyword(s):  
In Situ Hybridization ◽  
Genetic Regulation ◽  
Growth Control ◽  
Homeobox Genes ◽  
Blastema Cell ◽  
Genes Expression ◽  
Fin Ray ◽  
Msx Genes ◽  
Fin Development

To study the genetic regulation of growth control and pattern formation during fin development and regeneration, we have analysed the expression of four homeobox genes, msxA, msxB, msxC and msxD in zebrafish fins. The median fin fold, which gives rise to the unpaired fins, expresses these four msx genes during development. Transcripts of the genes are also present in cells of the presumptive pectoral fin buds. The most distal cells, the apical ectodermal ridge of the paired fins and the cleft and flanking cells of the median fin fold express all these msx genes with the exception of msxC. Mesenchymal cells underlying the most distal cells express all four genes. Expression of the msx genes in the fin fold and fin buds is transient and, by 3 days after fertilization, msx expression in the median fin fold falls below levels detectable by in situ hybridization. Although the fins of adult zebrafish normally have levels of msx transcripts undetectable by in situ hybridization, expression of all four genes is strongly reinduced during regeneration of both paired and unpaired fins. Induction of msx gene expression in regenerating caudal fins occurs as early as 30 hours postamputation. As the blastema forms, the levels of expression increase and reach a maximum between the third and fifth days. Then, msx expression progressively declines and disappears by day 12 when the caudal fin has grown back to its normal size. In the regenerating fin, the blastema cells that develop at the tip of each fin ray express msxB and msxC. Cells of the overlying epithelium express msxA and msxD, but do not express msxB or msxC. Amputations at various levels along the proximodistal axis of the fin suggest that msxB expression depends upon the position of the blastema, with cells of the rapidly proliferating proximal blastema expressing higher levels than the cells of the less rapidly proliferating distal blastema. Expression of msxC and msxD is independent of the position of the blastema cell along this axis. Our results suggest distinct roles for each of the four msx genes during fin development and regeneration and differential regulation of their expression.

Expression of homeobox gene Hox 1.1 during mouse embryogenesis

Development ◽  
1988 ◽  
Vol 104 (Supplement) ◽  
pp. 187-195 ◽  
Author(s):  
Kathleen A. Mahon ◽  
Heiner Westphal ◽  
Peter Gruss
Keyword(s):  
In Situ Hybridization ◽  
Homeobox Genes ◽  
Homeobox Gene ◽  
Spinal Ganglia ◽  
Spatial And Temporal Patterns ◽  
Rna Transcripts ◽  
Positional Identity ◽  
Specific Manner ◽  
Dorsal Spinal

Many of the genes controlling segmentation and pattern formation in Drosophila contain a conserved 183 bp sequence known as the homeobox. Homeobox sequences have been found in a range of metazoan species, including the vertebrates mouse and man. This striking conservation suggests that homeobox genes may play a fundamental role in developmental processes. If this is the case then it might be expected that vertebrate homeobox genes will be differentially expressed during embryogenesis and that the timing of their expression will coincide with major morphogenetic events. Here the spatial and temporal patterns of expression of murine homeobox genes will be explored, concentrating on the Hox 1.1 gene as an example. Using in situ hybridization to localize RNA transcripts, it has been found that Hox 1.1 is expressed in a region-specific manner during the formation and differentiation of the embryonic anteroposterior axis. Although striking patterns of expression of Hox 1.1 and other homeobox genes are seen in overtly segmented structures of the embryo (i.e. somites, prevertebral elements, neural tube and dorsal spinal ganglia) expression is also seen in tissues with no obvious segmental origin. The results suggest that homeobox genes probably do not play an exclusive role in segmentation in vertebrates, but are consistent with a role in the assignment of positional identity along the axis of the embryo.

In situ hybridization detection of homeobox genes reveals distinct expression patterns in oral squamous cell carcinomas

2011 ◽  
Vol 58 (2) ◽  
pp. 225-233 ◽  
Author(s):  
Tatiana N Libório ◽  
Thais Acquafreda ◽  
Luciana F Matizonkas-Antonio ◽  
Maria G Silva-Valenzuela ◽  
Alberto R Ferraz ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Squamous Cell ◽  
Expression Patterns ◽  
Homeobox Genes ◽  
Squamous Cell Carcinomas ◽  
Oral Squamous Cell Carcinomas ◽  
Hybridization Detection

The three most downstream genes of the Hox-3 cluster are expressed in human extraembryonic tissues including trophoblast of androgenetic origin

Development ◽  
1990 ◽  
Vol 108 (3) ◽  
pp. 471-477
Author(s):  
C.B. Oudejans ◽  
M. Pannese ◽  
A. Simeone ◽  
C.J. Meijer ◽  
E. Boncinelli
Keyword(s):  
In Situ Hybridization ◽  
Stromal Cells ◽  
Homeobox Genes ◽  
First Trimester ◽  
Splicing Regulation ◽  
Gene Promoters ◽  
Alternative Transcripts ◽  
Rna Probes ◽  
Extraembryonic Tissues

Human first trimester extraembryonic tissues of normal and androgenetic origin (molar pregnancies) were investigated for the expression of 6 homeobox genes from the chromosome 12-encoded Hox-3 cluster by non-autoradiographic in situ hybridization with biotinylated RNA probes. By comparative in situ hybridization involving the use of exon- or region-specific RNA probes, analysis included the cellular distribution of alternative Hox-3 transcripts in chorionic villous tissues. A bias in extraembryonic distribution was seen between transcripts of the three most upstream Hox-3 genes (Hox-3.7, -3.6, and -3.1) versus transcripts of the 3 most downstream genes (Hox-3.3, 3.4, and 3.5). Only genes from the latter group are transcribed in human extraembryonic tissues including extraembryonic tissues of androgenetic origin. Moreover, comparative in situ hybridization showed that distinct alternative transcripts of Hox-3.3, Hox 3.4 and Hox-3.5 are exclusively found in trophoblast cells while others are present in chorionic villous stromal cells as well. These data demonstrate the existence of tissue- and cell-specific use of transcriptional (alternative gene promoters) or post-transcriptional (alternative splicing) regulation of homeobox genes in extraembryonic tissues.

scholarly journals Isolation, Characterization, and in Situ Hybridization Studies of the Abundantly Transcribed Potato (Solanum tuberosum `Superior') Homeobox cDNA POTH1

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 453A-453
Author(s):  
Jennifer K. Hart ◽  
David J. Hannapel
Keyword(s):  
In Situ Hybridization ◽  
Potato Plant ◽  
Messenger Rna ◽  
Developmental Process ◽  
Homeobox Genes ◽  
Homeobox Gene ◽  
Vascular Cambium ◽  
Northern Analysis ◽  
Root Tips

Homeobox genes contain sequences coding for DNA-binding motifs. These sequences are highly conserved across both the animal and plant kingdoms. Members of this gene family code for transcription factors that are key regulators of developmental organization. In an attempt to further elucidate the developmental process of tuberization in the potato plant, a full-length homeobox cDNA has been isolated via sequence homology from an early tuberization stage cDNA library constructed from 4-day axillary bud tubers. This cDNA, POTH1, has been sequenced and characterized by Southern blotting, northern analysis, sequence comparison, and in situ hybridization. POTH1 is shown to be a class I homeobox gene with 45% overall similarity to Kn-1 of maize and 73% match in the homeobox region. Messenger RNA accumulation studies indicate that POTH1 mRNA, unlike most homeobox transcripts, is not limited to a particular organ or developmental stage. Instead, POTH1 mRNA accumulates in rapidly growing cells of the potato plant: the apical meristems, the vascular cambium, the edges of young leaves, axillary buds, and root tips. In situ studies indicate accumulation of POTH1 mRNA in the tunica and corpus layers of the apical dome of the shoot apex and the stolon apex. In the stolon, growth and proliferation of the parenchymal cells associated with the vascular cambium contribute to swelling during early stages of tuberization, and this tissue accumulates POTH1 mRNA. It is possible that POTH1 may be posttranscriptionally regulated in a particular organ or stage of growth, or that it is involved in a wider range of growth processes than most plant homeobox genes.

Genomic organization and expression of the planarian homeobox genes Dth-1 and Dth-2

Development ◽  
1993 ◽  
Vol 118 (1) ◽  
pp. 241-253 ◽  
Author(s):  
J. Garcia-Fernandez ◽  
J. Baguna ◽  
E. Salo
Keyword(s):  
Spatial Distribution ◽  
In Situ Hybridization ◽  
Genomic Organization ◽  
Evolutionary Relationship ◽  
Homeobox Genes ◽  
Genomic Analysis ◽  
Formation Mechanisms ◽  
Specific Cell ◽  
Specific Expression

We have characterized the genomic organization of Dth-1 and Dth-2, planarian homeobox-containing genes, previously described at the cDNA level (J. Garcia-Fernandez, J. Baguna and E. Salo (1991), Proc. Natl. Acad. Sci. USA, 88, 7338–7342). Genomic analysis shows that Dth-1 and Dth-2 genes encode proteins of 533 and 363 amino acids respectively. The open reading frame of Dth-1 is interrupted by two large introns of 8 kb and 12 kb Dth-2 also shows two introns, but these are short (42 bp and 44 bp) and the second interrupts helix III at position 44–45, as is the case with other homeobox genes from such divergent animals as Drosophila, honeybee, C. elegans, ascidians, and mouse, which suggests an ancient evolutionary relationship between these genes. The spatial distribution of transcripts in adult tissues, determined by in situ hybridization, demonstrates that Dth-1 is expressed at a high level in the gastrodermal cells, while Dth-2 is expressed in the peripheral parenchyma, at higher levels in the dorsal than the ventral regions. Their specific spatial distribution suggests a possible role for these homeobox genes in determination and/or differentiation of specific cell types. The expression pattern of both genes is more or less continuous, but in Dth-1 clustered discontinuous labelling in areas surrounding the gastrodermis may indicate a specific expression of this gene in groups of undifferentiated cells (neoblasts) already committed or determined to gastrodermal cell fates. In situ hybridization analysis during early regeneration shows expression only in the postblastema (stump) differentiated areas while no expression has been detected in the undifferentiated blastema, indicating that neither gene has a role in pattern formation mechanisms known to occur at the early stages of regeneration (0-3 days). Hence, Dth-1 and Dth-2 are planarian homeobox genes presumably involved in specific cell or tissue determination and/or differentiation.

scholarly journals Analysis of viral infections by in situ hybridization.

1986 ◽  
Vol 34 (1) ◽  
pp. 27-32 ◽  
Author(s):  
A T Haase
Keyword(s):  
In Situ Hybridization ◽  
Viral Infections ◽  
Single Cells ◽  
Genetic Regulation ◽  
Molecular Ecology ◽  
Simultaneous Detection ◽  
Real Life ◽  
Life Cycles ◽  
Hybridization Technique

In situ hybridization provides a versatile means of analysis of the life cycles of viruses in single cells. This kind of analysis in "real life" situations has provided considerable insight into the spread of viruses, mechanisms of tissue damage by viruses, and virus-host cell interactions in chronic diseases. In this article I describe refinements in technology underlining these advances, especially developments that have made the technique such a sensitive and quantitative one. I also describe a method for combined macroscopic and microscopic in situ hybridization, new assays for the simultaneous detection of genes and gene products in a single cell, and a double-label in situ hybridization technique. These methods have already proved useful in analyzing the molecular ecology of viral infections, and should find wide application to problems of genetic regulation in many other systems.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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