Expression of growth hormone-releasing hormone in human primary endometrial carcinomas

BACKGROUND: Hypothalamic GH-releasing hormone (GHRH) regulates GH release from the pituitary, but an ectopic production of GHRH has been detected in various non-hypothalamic tissues, especially cancers. OBJECTIVE: To investigate whether endometrial tumors produce GHRH. METHODS: Twenty-four endometrioid, three serous papillary (SP), three mixed type endometrioid/serous papillary adenocarcinomas and one malignant mixed Mullerian tumor (MMMT) were assessed for GHRH immunoreactivity by the polyclonal anti-rabbit antibody SV95 and for the expression of GHRH mRNA by in situ hybridization using an oligonucleotide probe. RESULTS: Increased GHRH immunoreactivity was detected in 15 out of 24 (63%) of the endometrioid tumors, including two out of three (66%) of the mixed type endometrioid/serous adenocarcinomas but not in the three SP or the MMMT tumor. Cytoplasmic staining was detected in all positive cases, while in three of them strong nuclear localization of GHRH was also revealed. In situ hybridization indicated the presence of GHRH mRNA in six cases, all characterized as positive for GHRH immunoreactivity. CONCLUSION: GHRH is expressed in a subset of endometrial tumors, of the endometrioid type in particular. A paracrine/autocrine role for GHRH in the development of the disease should be considered.

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Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

Journal of Histochemistry & Cytochemistry ◽

10.1177/42.9.8064134 ◽

1994 ◽

Vol 42 (9) ◽

pp. 1271-1276 ◽

Cited By ~ 14

Author(s):

M Numata ◽

T Ono ◽

S Iseki

Keyword(s):

In Situ Hybridization ◽

Significant Role ◽

Meiotic Prophase ◽

Oligonucleotide Probe ◽

Mouse Testis ◽

Seminiferous Tubules ◽

Hybridization Histochemistry ◽

In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

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Counting and Size Classification of Active Soil Bacteria by Fluorescence In Situ Hybridization with an rRNA Oligonucleotide Probe

Applied and Environmental Microbiology ◽

10.1128/aem.65.4.1753-1761.1999 ◽

1999 ◽

Vol 65 (4) ◽

pp. 1753-1761 ◽

Cited By ~ 109

Author(s):

Henrik Christensen ◽

Michael Hansen ◽

Jan Sørensen

Keyword(s):

In Situ Hybridization ◽

Nucleotide Sequence ◽

Fluorescence In Situ Hybridization ◽

Soil Bacteria ◽

Oligonucleotide Probe ◽

A Value ◽

Nonspecific Staining ◽

Dry Soil ◽

Active Bacteria

ABSTRACT A fluorescence in situ hybridization (FISH) technique based on binding of a rhodamine-labelled oligonucleotide probe to 16S rRNA was used to estimate the numbers of ribosome-rich bacteria in soil samples. Such bacteria, which have high cellular rRNA contents, were assumed to be active (and growing) in the soil. Hybridization to an rRNA probe, EUB338, for the domain Bacteria was performed with a soil slurry, and this was followed by collection of the bacteria by membrane filtration (pore size, 0.2 μm). A nonsense probe, NONEUB338 (which has a nucleotide sequence complementary to the nucleotide sequence of probe EUB338), was used as a control for nonspecific staining. Counting and size classification into groups of small, medium, and large bacteria were performed by fluorescence microscopy. To compensate for a difference in the relative staining intensities of the probes and for binding by the rhodamine part of the probe, control experiments in which excess unlabelled probe was added were performed. This resulted in lower counts with EUB338 but not with NONEUB338, indicating that nonspecific staining was due to binding of rhodamine to the bacteria. A value of 4.8 × 108 active bacteria per g of dry soil was obtained for bulk soil incubated for 2 days with 0.3% glucose. In comparison, a value of 3.8 × 108 active bacteria per g of dry soil was obtained for soil which had been air dried and subsequently rewetted. In both soils, the majority (68 to 77%) of actively growing bacteria were members of the smallest size class (cell width, 0.25 to 0.5 μm), but the active (and growing) bacteria still represented only approximately 5% of the total bacterial population determined by DAPI (4′,6-diamidino-2-phenylindole) staining. The FISH technique in which slurry hybridization is used holds great promise for use with phylogenetic probes and for automatic counting of soil bacteria.

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A Simple and Reliable Technique to Combine Oligonucleotide Probe in Situ Hybridization with Neuronal Tract Tracing in Vertebrate Embryos

Biotechnic & Histochemistry ◽

10.3109/10520299609117177 ◽

1996 ◽

Vol 71 (6) ◽

pp. 289-294 ◽

Cited By ~ 1

Author(s):

Bernd Fritzsch ◽

Finn Hallböök

Keyword(s):

In Situ Hybridization ◽

Oligonucleotide Probe ◽

Tract Tracing ◽

Reliable Technique ◽

Vertebrate Embryos

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Immunoautoradiographic and in situ hybridization analysis of corticotropin-releasing hormone biosynthesis in the hypothalamic paraventricular nucleus

Journal of Chemical Neuroanatomy ◽

10.1016/0891-0618(96)00124-x ◽

1996 ◽

Vol 11 (1) ◽

pp. 49-56 ◽

Cited By ~ 17

Author(s):

James P. Herman ◽

David G. Morrison

Keyword(s):

In Situ Hybridization ◽

Paraventricular Nucleus ◽

Hybridization Analysis ◽

Hypothalamic Paraventricular Nucleus ◽

Corticotropin Releasing Hormone ◽

Releasing Hormone ◽

Hormone Biosynthesis

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Distribution of thyrotropin-releasing hormone receptor messenger RNA in the rat brain: An in situ hybridization study

Neuroscience ◽

10.1016/0306-4522(92)90528-a ◽

1992 ◽

Vol 51 (4) ◽

pp. 891-909 ◽

Cited By ~ 56

Author(s):

L. Calzá ◽

L. Giardino ◽

S. Ceccatelli ◽

M. Zanni ◽

R. Elde ◽

...

Keyword(s):

In Situ Hybridization ◽

Rat Brain ◽

Hormone Receptor ◽

Messenger Rna ◽

Thyrotropin Releasing Hormone ◽

Hybridization Study ◽

Releasing Hormone

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Expression of Catalase mRNA and Protein in Adult Rat Brain: Detection by Nonradioactive In Situ Hybridization with Signal Amplification by Catalyzed Reporter Deposition (ISH–CARD) and Immunohistochemistry (IHC)/Immunofluorescence (IF)

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540305100606 ◽

2003 ◽

Vol 51 (6) ◽

pp. 751-760 ◽

Cited By ~ 30

Author(s):

Arno Schad ◽

H. Dariush Fahimi ◽

Alfred Völkl ◽

Eveline Baumgart

Keyword(s):

In Situ Hybridization ◽

Rat Brain ◽

Mammalian Cells ◽

Laser Scanning Microscopy ◽

Marker Enzyme ◽

Double Labeling ◽

Cytoplasmic Staining ◽

Catalyzed Reporter Deposition ◽

Nonradioactive In Situ Hybridization

Catalase, the classical peroxisomal marker enzyme, decomposes hydrogen peroxide and is involved in the antioxidant defense mechanisms of mammalian cells. In addition, catalase can oxidize, by means of its peroxidatic activity, a variety of substrates such as methanol and ethanol, producing the corresponding aldehydes. The involvement of brain catalase in the oxidation of ethanol is well established, and severe afflictions of the CNS in hereditary peroxisomal diseases (e.g., Zellweger syndrome) are well known. Whereas the distribution of catalase in the CNS has been investigated by enzyme histochemistry and immunohistochemistry (IHC), very little is known about the exact localization of catalase mRNA in brain. Here we report the application of a tyramine/CARD (catalyzed reporter deposition)-enhanced nonradioactive in situ hybridization (ISH) protocol for detection of catalase mRNA in sections of perfusion-fixed, paraffin-embedded rat brain. Catalase mRNA could be demonstrated in a large number of neurons throughout the rat brain as a distinct cytoplasmic staining signal with excellent morphological resolution. Compared to our standard ISH protocol, the CARD-enhanced protocol for catalase mRNA detection in rat brain showed higher sensitivity and significantly better signal-to-noise ratio. In parallel IHC experiments, using an antigen retrieval method consisting of combined trypsin digestion and microwave treatment of paraffin sections, the catalase antigen was found as distinct cytoplasmic granules in most catalase mRNA-positive neurons. In addition, catalase-positive granules, presumably peroxisomes, were found by confocal laser scanning microscopy in glial cells, which were identified by double labeling immunofluorescence for GFAP and CNPase for astroglial cells and oligodentrocytes, respectively. The excellent preservation of morphology and sensitive detection of both mRNA and protein in our preparations warrant the application of the protocols described here for systematic studies of catalase and other peroxisomal proteins in diverse pathological conditions such as Alzheimer's disease and aging.

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