Immunoreactivity of glutamate in mouse retina inner segment of photoreceptors with in vivo cryotechnique.
The purpose of this study was to clarify a previously controversial issue concerning glutamate (Glu) immunoreactivity (IR) in the inner segment (IS) of photoreceptors by using in vivo cryotechnique (IVCT) followed by freeze substitution (FS), which enabled us to analyze the cells and tissues reflecting living states. Eyeballs from anesthetized mice were directly frozen using IVCT. The frozen tissues were processed for FS fixation in acetone containing chemical fixatives, and embedded in paraffin. Deparaffinized sections were immunostained with an anti-Glu antibody. The strongest Glu-IR was obtained in the specimens prepared by FS with paraformaldehyde or a low concentration of glutaraldehyde, whereas no Glu-IR was obtained without the chemical fixatives. The Glu was immunolocalized in the IS, outer and inner plexiform and ganglion cell layers. Thus, the immunolocalization of Glu in the IS was clearly demonstrated using IVCT.
Terada
Nobuo
N
Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo-city, Yamanashi 409-3898, Japan. nobuot@yamanashi.ac.jp
Ohno
Nobuhiko
N
Saitoh
Sei
S
Saitoh
Yurika
Y
Ohno
Shinichi
S
D000818:Animals; D005404:Fixatives; D005557:Formaldehyde; D017110:Freeze Substitution; D018698:Glutamic Acid / Q000378:metabolism*; D005976:Glutaral; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D016612:Paraffin Embedding; D011108:Polymers; D055211:Retinal Photoreceptor Cell Inner Segment / Q000378:metabolism*; D016707:Tissue Fixation
D005404:Fixatives; D011108:Polymers; D005557:Formaldehyde; D018698:Glutamic Acid; D005976:Glutaral; C003043:paraform