A Confocal Laser Scanning Microscopic Study on the Distribution and fluorescent intensity of Calcitonin Gene-Related Peptide Immunoreactive Cells in the Trigeminal Ganglion of Aged Rats.
- Author:
Myung Kook KIM
1
Author Information
1. Department of Oral Anatomy, College of Dentistry, Seoul National University, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Calcitonin gene-related peptide;
Trigeminal ganglion cell;
Aging;
Rat;
Fluorescent intensity;
Confocal laser scannng microscopy
- MeSH:
Aging;
Animals;
Argon;
Calcitonin Gene-Related Peptide*;
Calcitonin*;
Fluorescence;
Imaging, Three-Dimensional;
Immunoglobulin G;
Microscopy, Confocal;
Rats*;
Rats, Sprague-Dawley;
Trigeminal Ganglion*
- From:Korean Journal of Anatomy
2000;33(2):191-200
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
The purpose of this study was to investigate the distribution and fluorescent intensity of the calcitonin gene-related peptide (CGRP) -immunoreactive cell bodies in the trigeminal ganglion of aged rats by means of confocal laser scanning microscopy. Eight control (3 months) and 8 aged (36 months) Sprague-Dawley rats were used. Trigeminal ganglion was removed and immersed in the 4% paraformaldehyde in 0.01 M phosphate buffer (pH 7.4). Serial frozon 30 micrometer thick sections were cut on cryostat. The rabbit antihuman CGRP was used as a primary antibody (1 : 2,000 in 0.01 M PBS at 4 degrees C). The fluorescence isothiocyanate (FITC) -conjugated anti-rabbit IgG was used as a secondary antibody (1 : 1,000 in 0.01 M PBS). The slides were observed under a Carl Zeiss LSM410 confocal laser scanning microscopy. Samples were excited by a 488 nm argon laser, and the images were filtered by a longpass 515 nm filter. Three-dimensional images were constructed from 16 serial images (each 1 micrometer thick) made by automatic optical sectioning. Unprocessed optical sections with an image size of 512X512 pixels and a pixel size of 0.2 micrometer were obtained and stored on a hard disk or optical disk. Color picture were printed on a video copy processor (Mitsubishi, Japan). The results were as follows; 1. The number of the CGRP-immunoreactive cell bodies in the trigeminal ganglion was 28.6 in the control rat and 19.4 in the aged rat. 2. In the optical single section, red (high fluorescent intensity) CGRP-immunoreactive cell bodies were the most abundant, followed by white (moderate fluorescent intensity), gray (low fluorescent intensity) in descending order in the control rat, whereas white CGRP-immunoreactive cell bodies were the most abundant, then gray, and red in desending order, in the aged rat. 3. In optical serial sections, red (high fluorescent intensity) CGRP-immunoreactive cell bodies were observed in all of the 16 sections of the control rat, whereas they were observed in a part of the 16 sections of the aged rat.
