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.

Allele-specific in situ hybridization (ASISH) analysis: a novel technique which resolves differential allelic usage of H19 within the same cell lineage during human placental development

Development ◽  
1996 ◽  
Vol 122 (3) ◽  
pp. 839-847 ◽  
Author(s):  
G.I. Adam ◽  
H. Cui ◽  
S.J. Miller ◽  
F. Flam ◽  
R. Ohlsson
Keyword(s):  
In Situ Hybridization ◽  
First Trimester ◽  
Paternal Allele ◽  
Monoallelic Expression ◽  
Hybridization Technique ◽  
Placental Development ◽  
Foetal Development ◽  
Biallelic Expression ◽  
Normal Human

Precursory studies of H19 transcription during human foetal development have demonstrated maternally derived monoallelic expression. Analyses in extra-embryonic tissues, however, have been more equivocal, with discernible levels of expression of the paternal allele of H19 documented in the first trimester placenta. By refining the in situ hybridization technique we have developed an assay to enable the functional imprinting status of H19 to be determined at the cellular level. This assay involves the use of oligonucleotide DNA probes that are able to discriminate between allelic RNA transcripts containing sequence polymorphisms. Biallelic expression of H19 is confined to a subpopulation of cells of the trophoblast lineage, the extravillous cytotrophoblast, while the mesenchymal stroma cells maintain the imprinted pattern of monoallelic expression of H19 throughout placental development. This data demonstrates that the low level of paternal H19 expression previously detected in normal human placenta is not due to a random loss of functional imprinting, but appears to result from a developmentally regulated cell type-specific activation of the paternal allele. In addition, biallelic expression of H19 does not seem to affect the functional imprinting of the insulin-like growth factor II gene, which is monoallelically expressed at relatively high levels in the extra-villous cytotrophoblasts. These results imply that the allelic usage of these two genes in normal human placental development may not be directly analogous to the situation previously documented in the mouse embryo.

In situ hybridization reveals differential spatial distribution of mRNAs for type I and type II collagen in the chick limb bud

Development ◽  
1988 ◽  
Vol 103 (1) ◽  
pp. 111-118 ◽  
Author(s):  
C.J. Devlin ◽  
P.M. Brickell ◽  
E.R. Taylor ◽  
A. Hornbruch ◽  
R.K. Craig ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Limb Development ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Limb Bud ◽  
Type I ◽  
Type Ii ◽  
Mrna Accumulation ◽  
Rna Probes

During limb development, type I collagen disappears from the region where cartilage develops and synthesis of type II collagen, which is characteristic of cartilage, begins. In situ hybridization using antisense RNA probes was used to investigate the spatial localization of type I and type II collagen mRNAs. The distribution of the mRNA for type II collagen corresponded well with the pattern of type II collagen synthesis, suggesting control at the level of transcription and mRNA accumulation. In contrast, the pattern of mRNA for type I collagen remained more or less uniform and did not correspond with the synthesis of the protein, suggesting control primarily at the level of translation or of RNA processing.

scholarly journals Localization of cells producing erythropoietin in murine liver by in situ hybridization [see comments]

Blood ◽  
1991 ◽  
Vol 77 (11) ◽  
pp. 2497-2503 ◽  
Author(s):  
ST Koury ◽  
MC Bondurant ◽  
MJ Koury ◽  
GL Semenza
Keyword(s):  
In Situ Hybridization ◽  
Transgenic Mice ◽  
Antisense Rna ◽  
Interstitial Cells ◽  
Cell Type ◽  
Isolated Cells ◽  
Rna Probes ◽  
Sense Strand ◽  
Murine Liver

Abstract In situ hybridization using antisense RNA probes was used to localize cells that produce erythropoietin (EPO) in the livers of anemic transgenic mice expressing the human EPO gene and in livers of anemic nontransgenic mice. In transgenic mice bled from a hematocrit of 55% to one of 10%, hepatocytes surrounding central veins synthesized large amounts of human EPO mRNA. EPO-producing cells were very rare in the area of portal triads. In transgenic mice bled to a hematocrit of 20%, a similar number and distribution of cells contained human EPO mRNA as was found with a 10% hematocrit, but the cells were less heavily labeled, indicating increased EPO production per cell at 10% hematocrit as compared with 20% hematocrit. No human EPO mRNA was detected in the kidneys of anemic transgenic mice, although endogenous murine EPO mRNA was strongly expressed in cortical interstitial cells. In sections of livers from nontransgenic mice bled from a hematocrit of 45% to one of 10%, only isolated cells produced EPO. When the types of cells could clearly be identified, approximately 80% of these cells were hepatocytes, while 20% had a nonepithelial morphology and were located in or adjacent to the sinusoidal spaces. When the sense strand was used as the RNA probe for in situ hybridization, no labeled cells were seen in normal or anemic livers. These results demonstrate that hepatocytes are responsible for production of EPO in both transgenic and nontransgenic mice and that a second cell type that is similar in morphology to EPO-producing interstitial cells in the kidney also produces EPO in the livers of nontransgenic mice.

In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes

1986 ◽  
Vol 18 (11-12) ◽  
pp. 597-604 ◽  
Author(s):  
Heinz Hoefler ◽  
Henry Childers ◽  
Marc R. Montminy ◽  
Ronald M. Lechan ◽  
Richard H. Goodman ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Antisense Rna ◽  
Tissue Sections ◽  
Rna Probes

Localization of progesterone-associated endometrial protein mRNA by in-situ hybridization in human pregnancy decidua, endometriosis and borderline endometrioid adenoma

1993 ◽  
Vol 10 (1) ◽  
pp. 71-77 ◽  
Author(s):  
M Kämäräinen ◽  
I Leivo ◽  
M Julkunen ◽  
M Seppälä
Keyword(s):  
In Situ Hybridization ◽  
Cellular Localization ◽  
First Trimester ◽  
Glandular Epithelium ◽  
Hybridization Histochemistry ◽  
Human Pregnancy ◽  
In Situ Hybridization Histochemistry

ABSTRACT Progesterone-associated endometrial protein (PAEP) has been isolated from human decidualized endometrium. In-situ hybridization histochemistry was employed to determine the cellular localization of PAEP mRNA in decidua during pregnancy. PAEP mRNA was found to be expressed in the glandular epithelium of decidua spongiosa throughout pregnancy. Substantial variations in the amount of PAEP mRNA during the course of pregnancy were observed, and it was most abundant at the end of the first trimester. We also found that the PAEP gene was expressed in endometriosis and in a borderline endometrioid adenoma. As in decidual tissues, PAEP mRNA in endometriosis was abundant in the glandular compartment.

scholarly journals PCR-derived ssDNA Probes for Fluorescent In Situ Hybridization to HIV-1 RNA

2000 ◽  
Vol 48 (2) ◽  
pp. 285-293 ◽  
Author(s):  
Marlyse C. Knuchel ◽  
Brigit Graf ◽  
Erika Schlaepfer ◽  
Herbert Kuster ◽  
Marek Fischer ◽  
...  
Keyword(s):  
T Cells ◽  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Uracil Dna Glycosylase ◽  
Rna Probes ◽  
Immunodeficiency Virus ◽  
Polymerase Chain ◽  
Rna Probe ◽  
Hiv 1

We developed a simple and rapid technique to synthesize single-stranded DNA (ssDNA) probes for fluorescent in situ hybridization (ISH) to human immunodeficiency virus 1 (HIV-1) RNA. The target HIV-1 regions were amplified by the polymerase chain reaction (PCR) and were simultaneously labeled with dUTP. This product served as template for an optimized asymmetric PCR (one-primer PCR) that incorporated digoxigenin (dig)-labeled dUTP. The input DNA was subsequently digested by uracil DNA glycosylase, leaving intact, single-stranded, digoxigenin-labeled DNA probe. A cocktail of ssDNA probes representing 55% of the HIV-1 genome was hybridized to HIV-1-infected 8E5 T-cells and uninfected H9 T-cells. For comparison, parallel hybridizations were done with a plasmid-derived RNA probe mix covering 85% of the genome and a PCR-derived RNA probe mix covering 63% of the genome. All three probe types produced bright signals, but the best signal-to-noise ratios and the highest sensitivities were obtained with the ssDNA probe. In addition, the ssDNA probe syntheses generated large amounts of probe (0.5 to 1 μg ssDNA probe per synthesis) and were easier to perform than the RNA probe syntheses. These results suggest that ssDNA probes may be preferable to RNA probes for fluorescent ISH.

In Situ Hybridization with Riboprobes: An Overview for Veterinary Pathologists

1998 ◽  
Vol 35 (3) ◽  
pp. 159-167 ◽  
Author(s):  
C. Brown
Keyword(s):  
In Situ Hybridization ◽  
Molecular Biology ◽  
Infectious Agents ◽  
Probe Target ◽  
Cellular Genes ◽  
Rna Probes ◽  
Pcr Technology ◽  
Labeled Rna ◽  
Target Combination

In situ hybridization using nonradioactively-labeled RNA probes is a technique that combines understanding of basic molecular biology and histopathologic interpretation. Recombinant or PCR technology can be used to produce probes that hybridize with a wide variety of cellular genes and infectious agents. Hybridization conditions can be optimized for each probe/target combination.

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