Chapter 15
Electron Microscopic in Situ Hybridization and
its Combination With Immunohistochemistry
AKIRA MATSUNO, TADASHI NAGASHIMA,
R. YOSHIYUKI OSAMURA AND KElleHI WATANABE
Introduction
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; Wolber 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 (Matsuno et al. 1996), which can
Correspondence to Akira Matsuno, Teikyo University Ichihara Hospital,
Department of Neurosurgery, 3426-3 Anegasaki, Ichihara City, Chiba,
299-0111, Japan (phone +81-436-62-1211;fax +81-436-62-1357; e-mail akirakun@med.teikyo-u.ac.jp); Tadashi Nagashima, Teikyo University Ichihara Hospital, Department of Neurosurgery, 3426-3 Anegasaki, Ichihara
City, Chiba, 299-0111, Japan; R. Yoshiyuki Osamura, Tokai University
School of Medicine, Department of Pathology, Boseidai, Isehara City, Kanagawa, 259-1100, Japan; Keiichi Watanabe, Tokai University School of
Medicine, Department of Pathology, Boseidai, Isehara City, Kanagawa,
259-1100, Japan
Electron Microscopic in Situ Hybridization and
its Combination With Immunohistochemistry
AKIRA MATSUNO, TADASHI NAGASHIMA,
R. YOSHIYUKI OSAMURA AND KElleHI WATANABE
Introduction
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; Wolber 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 (Matsuno et al. 1996), which can
Correspondence to Akira Matsuno, Teikyo University Ichihara Hospital,
Department of Neurosurgery, 3426-3 Anegasaki, Ichihara City, Chiba,
299-0111, Japan (phone +81-436-62-1211;fax +81-436-62-1357; e-mail akirakun@med.teikyo-u.ac.jp); Tadashi Nagashima, Teikyo University Ichihara Hospital, Department of Neurosurgery, 3426-3 Anegasaki, Ichihara
City, Chiba, 299-0111, Japan; R. Yoshiyuki Osamura, Tokai University
School of Medicine, Department of Pathology, Boseidai, Isehara City, Kanagawa, 259-1100, Japan; Keiichi Watanabe, Tokai University School of
Medicine, Department of Pathology, Boseidai, Isehara City, Kanagawa,
259-1100, Japan