In situ hybridization of myoglobin mRNA: results on the skeletal muscles of normal subjects and patients with neuromuscular diseases

1993 ◽  
Vol 86 (4) ◽  
pp. 313-318 ◽  
Author(s):  
Takao Mitsui ◽  
Hisaomi Kawai ◽  
Takako Naruo ◽  
Hiroshi Nishino ◽  
Shiro Saito
1992 ◽  
Vol 133 (1) ◽  
pp. 67-NP ◽  
Author(s):  
E. D. Schmidt ◽  
E. D. L. Schmidt ◽  
R. van der Gaag ◽  
R. Ganpat ◽  
L. Broersma ◽  
...  

ABSTRACT The correlation between the occurrence of Graves' ophthalmopathy and Graves' hyperthyroidism may indicate a role for tri-iodothyronine (T3) hormone in the pathogenesis of Graves' ophthalmopathy. In Graves' ophthalmopathy the recti eye muscles are greatly enlarged whereas skeletal muscles seem unaffected. The distribution of the nuclear T3 receptor was studied in normal human and rat eye and skeletal muscles with immunohistochemistry using mouse (monoclonal) antibodies, and by in-situ hybridization for the detection of mRNA encoding the T3-receptor protein. Nuclear staining with T3-receptor antibodies was found in all types of tissues studied. Cytoplasmic staining occurred predominantly in the muscle fibres of the orbital layer of the eye muscles and was generally absent or very low in skeletal muscle fibres and hepatocytes. Immunostaining could be inhibited by preabsorbing the antibodies with bacterially expressed T3-receptor protein, implying specificity. The presence of nuclear and cytoplasmic hormonefree T3 receptor sites was indicated after preincubation of sections with T3 hormone: T3-receptor immunostaining decreased and T3-hormone staining increased. In-situ hybridization clearly revealed the presence of α-1 and β-1 forms of the T3-receptor mRNA in liver, skeletal muscles, and orbital and intermediate layers of the eye muscles. The data demonstrate the presence of T3 hormone-receptor molecules in the extraocular and skeletal muscles. The different susceptibilities of these muscles to Graves' hyperthyroidism may relate to the quantitative differences in T3 hormone-receptor distribution. Journal of Endocrinology (1992) 133, 67–74


Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  

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.


Author(s):  
Gary Bassell ◽  
Robert H. Singer

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.


2007 ◽  
Vol 177 (4S) ◽  
pp. 596-597
Author(s):  
Joseph P. Alukal ◽  
Bobby B. Najari ◽  
Wilson Chuang ◽  
Lata Murthy ◽  
Monica Lopez-Perdomo ◽  
...  

2006 ◽  
Vol 175 (4S) ◽  
pp. 287-288 ◽  
Author(s):  
Juliann M. Dziubinski ◽  
Michael F. Sarosdy ◽  
Paul R. Kahn ◽  
Mark D. Ziffer ◽  
William R. Love ◽  
...  

2004 ◽  
Vol 171 (4S) ◽  
pp. 156-156
Author(s):  
Chandler D. Dora ◽  
Yasushi Kondo ◽  
Fusheng X. Lan ◽  
Jeffrey M. Slezak ◽  
Erik J. Bergstralh ◽  
...  

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