Electron Microscopic in Situ Hybridization and its Combination With Immunohistochemistry
In situ hybridization at the electron microscopic level is a useful method for examining the intracellular distribution and role of mRNA in protein synthesis. Ultrastructural in situ hybridization for mRNA has been carried out by three different approaches: preembedding method (Guitteny et al. 1989; W olber et al. 1989; Trembleau et al. 1990; Pomeroy et al. 1991; Matsuno et al. 1994a,b, 1995), ultrathin frozen sections (Morel et al. 1989a,b; Le Guellec et al. 1992) and postembedding method (Jirikowski et al. 1990; Le Guellec et al. 1990, 1991, 1992, Matsuno et al. 1994a, 1995). In recent years, we have developed non-radioisotopic electron microscopic in situ hybridization method using biotinylated synthesized oligonucleotide probes for the ultrastructural visualization of growth hormone (GH) and prolactin (PRL) mRNAs in rat pituitary cells, and have applied this method to pathophysiological studies of pituitary cells (Matsuno et al. 1994a,b, 1995). In addition, we have developed combined method of electron microscopic in situ hybridization and immunohistochemistry for simultaneous detection of pituitary hormone and its message in the same cell (Matsumo et al. 1996), which can serve the visualization of intracellular spatial relationship between pituitary hormone and its message at the ultrastructural level.
KeywordsRough Endoplasmic Reticulum Colloidal Gold Electron Microscopic Level Sodium Periodate Colloidal Gold Particle
Unable to display preview. Download preview PDF.
- Escaig-Haye F, Grigogiev V, Sharova I, Rudneva V, Buckrinskaya A, Fournier J.G. (1992) Ultrastructural localization of HIV-1 RNA and core proteins. Simultaneous visualization using double immunogold labelling after in situ hybridization and immunocytochemistry. J Submicrosc Cytol Pathol 24:437–443PubMedGoogle Scholar
- Gingras D, Bendayan M (1995) Colloidal gold electron microscopic in situ hybridization: combination with immunocytochemistry for the study of insulin and amylase secretion. Cell Vision 2:218–225Google Scholar
- Matsuno A, Ohsugi Y, Utsunomiya H, Takekoshi S, Osamura R.Y., Watanabe K, Teramoto A (1994a) Ultrastructural distribution of growth hormone, prolactin mRNA in normal rat pituitary cells: A comparison between preembedding and postembedding methods. Histochemistry 102:265–270PubMedCrossRefGoogle Scholar
- Matsuno A, Ohsugi Y, Utsunomiya H, Takekoshi S, Sanno N, Osamura R.Y., Watanabe K, Teramoto A, Kirino T (1995) Changes in the ultrastructural distribution of prolactin and growth hormone mRNAs in pituitary cells of female rats after estrogen and bromocriptine treatment, studied using in situ hybridization with biotinylated oligonucleotide probes. Histochem Cell Biol 104:37–45PubMedCrossRefGoogle Scholar
- Matsuno A, Teramoto A, Takekoshi S, Utsunomiya H, Ohsugi Y, Kishikawa S, Osamura R.Y., Kirino T, Lloyd R.V. (1994b) Application ofbiotinylated oligonucleotide probes to the detection of pituitary hormone mRNA using Northern blot analysis, in situ hybridization at light and electron microscopic levels. Histochem J 26:771–777PubMedGoogle Scholar
- Matsuno A, Utsunomiya H, Ohsugi Y, Takekoshi S, Sanno N, Osamura R.Y.Google Scholar