scholarly journals Simultaneous detection of two messenger RNAs in the central nervous system: a simple two-step in situ hybridization procedure using a combination of radioactive and non-radioactive probes.

1991 ◽  
Vol 39 (11) ◽  
pp. 1575-1578 ◽  
Author(s):  
E Normand ◽  
B Bloch
Keyword(s):  
Central Nervous System ◽  
In Situ Hybridization ◽  
Simultaneous Detection ◽  
Messenger Rnas ◽  
Tissue Sections ◽  
System A ◽  
Step Procedure ◽  
Hybridization Procedure ◽  
The Central Nervous System

We present here a method enabling the simultaneous detection of two messenger RNAs in tissue sections by use of a two-step in situ hybridization procedure. Tissue sections were hybridized with a radioactive probe and coated with emulsion. The emulsion was processed for development, fixed, and a second hybridization was performed through the emulsion with a biotinylated probe subsequently revealed with streptavidin-alkaline phosphatase. This procedure allows the detection of two mRNAs without loss of signal, removal of the emulsion, or spurious reaction. The simultaneous detection of oxytocin and vasopressin mRNAs in the hypothalamus, and of dopamine receptor and neuropeptide mRNAs in the striatum, demonstrated the efficiency of the procedure. Such a two-step procedure provides a simple and flexible way to make possible comparative analysis of the localization of two mRNAs within the same tissue section.

Quest for a reliable, valid, and sensitive in situ hybridization procedure to detect viral nucleic acids in the central nervous system

1987 ◽  
Vol 12 (6) ◽  
pp. 565-571 ◽  
Author(s):  
Wallace W. Tourtellotte ◽  
Peter Schmid ◽  
Peter Pick ◽  
Neil Verity ◽  
Steven Martinez ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
In Situ Hybridization ◽  
Nucleic Acids ◽  
Hybridization Procedure ◽  
The Central Nervous System

scholarly journals Co-localization of EGF transcripts and peptides by combined immunohistochemistry and in situ hybridization.

1990 ◽  
Vol 38 (12) ◽  
pp. 1853-1857 ◽  
Author(s):  
R I Couwenhoven ◽  
W Luo ◽  
M L Snead
Keyword(s):  
In Situ Hybridization ◽  
Growth Factor ◽  
Simultaneous Detection ◽  
Interstitial Cells ◽  
Glandular Epithelium ◽  
Messenger Rnas ◽  
Tissue Sections ◽  
Representative Model ◽  
Epidermal Growth

There is increasing evidence that autocrine- and paracrine-acting growth factors participate in cell and tissue development, maintenance, and renewal. Recent advances in histochemical techniques have facilitated the localization of growth factor messenger RNAs or polypeptides in tissue sections. However, the spatial relationships between the sites of growth factor transcription, translation, and post-translational processing to functional bioactive peptides have been difficult to correlate because each method of detection requires separate tissue sections. We undertook the simultaneous detection of epidermal growth factor (EGF) transcripts and EGF epitopes by combining immunohistochemistry methods with in situ hybridization. Adult mouse submandibular gland was chosen as a representative model because it contains sites of EGF biosynthesis which may participate in mediating the development, maintenance, and renewal of the organ through autocrine or paracrine mechanism(s). Granular duct (GD) cells demonstrated the presence of both EGF transcripts and EGF peptides. In contrast, the interstitial cells lying adjacent to glandular epithelium also contained relatively high levels of EGF transcripts, although no mature EGF peptides were detected. The experimental approach of co-localization and the resulting data indicate previously unreported sites of EGF transcription in glandular interstitial cells, which may provide molecular information required for the morphogenesis and differentiation of adjacent glandular epithelium.

scholarly journals N-myc proto-oncogene expression during organogenesis in the developing mouse as revealed by in situ hybridization.

1988 ◽  
Vol 107 (4) ◽  
pp. 1325-1335 ◽  
Author(s):  
G Mugrauer ◽  
F W Alt ◽  
P Ekblom
Keyword(s):  
In Situ Hybridization ◽  
Hair Follicles ◽  
Early Differentiation ◽  
Cell Lineages ◽  
Tissue Sections ◽  
Embryonic Tissues ◽  
Differentiated Cells ◽  
The Central Nervous System ◽  
Developing Kidney

The N-myc proto-oncogene is expressed during embryogenesis, suggesting that it plays a role in normal development. Since the myc-family oncogenes have been implicated in the control of cell growth, the embryonic expression may reflect rapid proliferation known to occur in development. Alternatively, N-myc expression may be involved in specific differentiation stages. In many embryonic tissues, early and late differentiation events occur in different locations. By in situ hybridization of tissue sections, we now demonstrate a restricted expression of N-myc mRNA to a few tissues and to areas where the first differentiation stages occur. N-myc expression was most strongly expressed in the developing kidney, hair follicles, and in various parts of the central nervous system. In these tissues, expression was restricted to a few cell lineages. In all lineages, expression was confined to early differentiation stages, and, at onset of overt differentiation, the level of expression decreased dramatically. Several rapidly proliferating tissues showed very little, if any, N-myc expression. In the brain, post-mitotic but not yet differentiated cells expressed high levels of N-myc mRNA. Therefore, N-myc expression is not a simple marker for proliferation in the embryo. Rather, N-myc expression seems to be a feature of early differentiation stages of some cell lineages in kidney, brain, and hair follicles, regardless of the proliferative status of the cell. The results raise the possibility that N-myc may participate in the control of these early differentiation events.

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