Localization of specific mRNA sequences in Xenopus laevis embryos by in situ hybridization

Development ◽  
1986 ◽  
Vol 91 (1) ◽  
pp. 153-168
Author(s):  
Eva Dworkin-Rastl ◽  
Darcy B. Kelley ◽  
Mark B. Dworkin
Keyword(s):  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Lateral Plate ◽  
Tissue Specific ◽  
Maternal Rna ◽  
Specific Hybridization ◽  
Cloned Cdna ◽  
Xenopus Laevis Embryos ◽  
Specific Sequences

In situ hybridization of cloned cDNA probes to frozen sections of Xenopus laevis stage-42 tadpoles has been used to determine the tissue localization of several mRNAs. Nine out of sixteen probes tested hybridized to most or all tadpole tissues; seven probes exhibited tissue-specific hybridization. The non-tissue-specific sequences hybridized to RNA species that are also present in maternal RNA while the tissue-specific sequences hybridized to embryonic RNA species induced after gastrulation and undetectable in maternal RNA. Tissue-specific hybridization was observed with muscle (five clones), epidermis (one clone), and the nervous system (one clone). All muscle-specific sequences hybridized to somites and lateral plate muscles, but they differed in their hybridization to heart muscle.

Region-specific variation of gene expression in the human epididymis as revealed by in situ hybridization with tissue-specific cDNAs

1993 ◽  
Vol 34 (1) ◽  
pp. 16-24 ◽  
Author(s):  
Nora Krull ◽  
Richard Ivell ◽  
Caroline Osterhoff ◽  
Christiane Kirchhoff
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Tissue Specific ◽  
Human Epididymis ◽  
Specific Variation

scholarly journals UNDER-REPLICATION OF RIBOSOMAL CISTRONS IN POLYTENE CHROMOSOMES OF RHYNCHOSCIARA

1974 ◽  
Vol 62 (1) ◽  
pp. 215-222 ◽  
Author(s):  
A. G. Gambarini ◽  
F. J. S. Lara
Keyword(s):  
Salivary Gland ◽  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Related Species ◽  
Polytene Chromosomes ◽  
Chromosome X ◽  
Heterologous Hybridization ◽  
Ribosomal Cistrons ◽  
Rhynchosciara Americana

DNA preparations obtained from several tissues of Rhynchosciara americana and two related species, R. milleri and R. papaveroi, were hybridized to R. americana rRNA. The percentage of hybridization was found to be higher in tissues with low polyteny than in tissues with high polyteny, suggesting a relationship between the amount of rDNA and the tissue polyteny. This could be explained by under-replication of ribosomal cistrons in polytene cells, such as those from the salivary gland. Only slight tissue-dependent changes in the percentages of hybridization can be observed in heterologous hybridization using Xenopus laevis rRNA. The possibility that these experiments could not detect differences in the amount of ribosomal cistrons among tissues is discussed. The female:male ratio for the percentages of hybridization in the salivary gland of R. americana agrees with the results obtained by in situ hybridization experiments (16, 17) which have shown that the rRNA cistrons are distributed among chromosomes other than chromosome X.

Localization of the sites of synthesis and action of prolactin by immunocytochemistry and in-situ hybridization within the human utero-placental unit

1991 ◽  
Vol 7 (3) ◽  
pp. 241-247 ◽  
Author(s):  
W.-X. Wu ◽  
J. Brooks ◽  
M. R. Millar ◽  
W. L. Ledger ◽  
P. T. K. Saunders ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Culture Medium ◽  
Cdna Probe ◽  
Specific Hybridization ◽  
Decidual Cells ◽  
Its Gene ◽  
First Time ◽  
Silver Grains ◽  
Prolactin Mrna

ABSTRACT While the fetal pituitary synthesizes and releases prolactin, it is also produced within the utero-placental unit during pregnancy in women and has been localized in the amnion, chorion and decidua. However, it is not clear whether prolactin is synthesized within all these non-fetal pituitary tissues. We have investigated prolactin production and its gene expression using tissue culture, immunocytochemistry and in-situ hybridization techniques. Prolactin was immunolocalized not only in the decidua but also in amnion and trophoblast cells. In contrast, the in-situ hybridization results showed that silver grains, formed by specific hybridization of a prolactin cDNA probe to prolactin mRNA, were confined to decidual cells of early and term pregnancy. The results from tissue cultures correlated well with those of in-situ hybridization, that is that only the decidua made detectable prolactin, while it was undetectable in the culture medium from trophoblast tissue, irrespective of the stage of pregnancy. This study, for the first time, establishes that only decidualized cells are involved in biosynthesis of prolactin; other prolactin-containing cells in the amnion and trophoblast appear to sequester prolactin, possibly via receptors, suggesting that prolactin may play an important paracrine role within the amnion and syncitio- and cytotrophoblast of the utero-placental unit.

In situ localization of mRNA for epidermal growth factor in the submandibular gland of the mouse.

1985 ◽  
Vol 33 (12) ◽  
pp. 1235-1240 ◽  
Author(s):  
E W Gresik ◽  
R M Gubits ◽  
T Barka
Keyword(s):  
In Situ Hybridization ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Submandibular Gland ◽  
Cdna Probe ◽  
Coding Region ◽  
Specific Hybridization ◽  
Epidermal Growth ◽  
Convoluted Tubules

Epidermal growth factor (EGF) is a polypeptide originally isolated from the mouse submandibular gland, where it is localized immunocytochemically in cells of the granular convoluted tubules (GCT). cDNAs encoding the precursor of mouse submandibular EGF have been cloned (Scott et al. Science 221:236, 1983; Gray et al. Nature 303:722, 1983). A fragment of one of these clones, pmegf10, containing the EGF coding region, was tritium-labeled by nick-translation and used as a probe for in situ hybridization to EGF mRNA. A specific hybridization signal for EGF mRNA was seen only in mature or developing GCT cells. The intensity of the signal was stronger in glands of intact males than in females or in castrated males. In glands of castrates treated with testosterone, or of intact females treated with triiodothyronine (T3), the signal was comparable to that in intact males. In glands of males treated with T3 the intensity of the signal was stronger than in untreated males. A weak to moderate signal was seen in developing GCT cells of 20-day-old males but not females. Hybridization for 3 days gave a stronger signal than that for 1 day. No signal was seen in either sex at 10 days of age, or in control preparations exposed to labeled DNA of pBR322. The presence of EGF mRNA exclusively in GCT cells provides strong evidence that these cells are the only site of synthesis of EGF in the submandibular gland. In situ hybridization with this cDNA probe will provide a sensitive method to determine possible cellular sites of EGF production outside of the submandibular gland.

The genomic composition of Tricepiro, a synthetic forage crop

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 154-159 ◽  
Author(s):  
María Rosa Ferrari ◽  
Eduardo J Greizerstein ◽  
Héctor A Paccapelo ◽  
Carlos A Naranjo ◽  
Angelines Cuadrado ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chromosomal Location ◽  
Hexaploid Triticale ◽  
Genomic Constitution ◽  
Forage Crop ◽  
A Genome ◽  
Total Dna ◽  
Synthetic Hexaploid ◽  
Specific Sequences

Chromosome in situ hybridization (FISH and GISH) is a powerful tool for determining the chromosomal location of specific sequences and for analysing genome organization and evolution. Tricepiro (2n = 6x = 42) is a synthetic cereal obtained by G. Covas in Argentina (1972), which crosses hexaploid triticale (2n = 6x = 42) and octoploid Trigopiro (2n = 8x = 56). Several years of breeding produced a forage crop with valuable characteristics from Secale, Triticum, and Thinopyrum. The aim of this work is to analyse the real genomic constitution of this important synthetic crop. In situ hybridization using total DNA of Secale, Triticum, and Thinopyrum as a probe (GISH) labelled with biotin and (or) digoxigenin showed that tricepiro is composed of 14 rye chromosomes and 28 wheat chromosomes. Small zones of introgression of Thinopyrum on wheat chromosomes were detected. The FISH using the rye repetitive DNA probe pSc 119.2 labelled with biotin let us characterize the seven pairs of rye chromosomes. Moreover, several wheat chromosomes belonging to A and B genomes were distinguished. Therefore, tricepiro is a synthetic hexaploid (2n = 6x = 42) being AABBRR in its genomic composition, with zones of introgression of Thinopyrum in the A genome of wheat.Key words: tricepiro, trihybrid, Triticum, Secale, Thinopyrum, in situ hybridization, FISH, GISH, genomic composition, synthetic forage crop.

Immunocytochemistry and in situ hybridization of neuropeptide Y in the hypothalamus of Xenopus laevis in relation to background adaptation

Neuroscience ◽  
1993 ◽  
Vol 55 (3) ◽  
pp. 667-675 ◽  
Author(s):  
R. Tuinhof ◽  
F.Y.S.C. Laurent ◽  
R.G.E. Ebbers ◽  
W.J.A.J. Smeets ◽  
M.C.H.M. van Riel ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Neuropeptide Y ◽  
Background Adaptation

Localization of low abundance DNA sequences in tissue sections by in situ hybridization

1986 ◽  
Vol 81 (1) ◽  
pp. 143-162 ◽  
Author(s):  
C.W. Lo
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Detection System ◽  
Mouse Tissue ◽  
Globin Genes ◽  
Beta Globin ◽  
Tissue Sections ◽  
Specific Hybridization ◽  
Hybridization Procedure

Specific DNA sequences were loaclized in the nuclei of paraffin-embedded mouse tissue sections with in situ hybridization using a biotinylated globin probe in conjunction with a streptavidin-alkaline phosphatase detection system. Globin inserts were clearly detected in the tissue sections of transgenic mice containing 1000, 120 or 5 copies of the exogenously introduced beta-globin genes. In addition, specific hybridization signal was also obtained for the endogenous complement of beta-globin genes in the tissue sections of normal mice. These results demonstrate that this hybridization procedure is very sensitive and should be useful for characterizing the distribution of low abundance DNA sequences in cells and tissue sections.

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