|
|
|
|
 |
|||
|
|||
Combination of non-radioactive and radioactive in situ hybridization with immunohistochemistry: a new method allowing the simultaneous detection of two mRNAs and one antigen in the same brain tissue section
A Trembleau, D Roche and A Calas
Department de Cytologie, Institut des Neurosciences CNRS URA 1488, Universite P. & M. Curie, Paris, France.
We describe here a simple method for combining non-radioactive and radioactive in situ hybridization and immunohistochemistry on the same brain tissue section. This approach was first developed on the well- characterized hypothalamo-neurohypophyseal system, facilitating the optimization of the triple-labeling procedure and the verification of labeling specificity. We report the simultaneous detection of vasopressin (VP) mRNA with a digoxigenin-labeled oligonucleotide, oxytocin (OT) mRNA with a 35S-labeled oligonucleotide, and OT peptide in the same 12-microns cryostat section. This was performed on floating sections as follows: first, the two probes were hybridized simultaneously; second, the peptide was detected with an immunoperoxidase-DAB procedure; third, the digoxigenin-labeled probe was detected with an alkaline phosphatase-NBT/BCIP technique; and finally, the 35S-labeled probe was detected by histological autoradiography. We also demonstrate that this approach is suitable for the simultaneous detection of tyrosine hydroxylase and two less abundant mRNAs, vasoactive intestinal peptide and vasopressin mRNAs, in the suprachiasmatic nucleus. The combination of the three techniques did not significantly diminish their specificity or sensitivity. In conclusion, this new method, permitting the simultaneous detection of three different products of gene expression in the same section, could be useful for further analysis of the phenotypic organization and its plasticity in endocrine or neural tissues.
Volume 41,
Issue 4,
pp. 489-498,
04/01/1993
Copyright © 1993 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S. L. Erlandsen, E. Jarroll, P. Wallis, and H. van Keulen Development of Species-specific rDNA Probes for Giardia by Multiple Fluorescent In Situ Hybridization Combined with Immunocytochemical Identification of Cyst Wall Antigens J. Histochem. Cytochem., August 1, 2005; 53(8): 917 - 927. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. E. Garcia, Y. Xia, J. Harrison, C. B. Wilson, R. J. Johnson, K. B. Bacon, and L. Feng Mononuclear Cell-Infiltrate Inhibition by Blocking Macrophage-Derived Chemokine Results in Attenuation of Developing Crescentic Glomerulonephritis Am. J. Pathol., April 1, 2003; 162(4): 1061 - 1073. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-M. Vacher, P. Fretier, C. Creminon, A. Calas, and H. Hardin-Pouzet Activation by Serotonin and Noradrenaline of Vasopressin and Oxytocin Expression in the Mouse Paraventricular and Supraoptic Nuclei J. Neurosci., March 1, 2002; 22(5): 1513 - 1522. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Grino and A. J. Zamora An In Situ Hybridization Histochemistry Technique Allowing Simultaneous Visualization by the Use of Confocal Microscopy of Three Cellular mRNA Species in Individual Neurons J. Histochem. Cytochem., June 1, 1998; 46(6): 753 - 760. [Abstract] [Full Text]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |