Single-cell imaging of c-fos expression in rat primary hippocampal cells using a luminescence microscope.
Methods to study gene expression in live cells over time have been limited. One known method is the luciferase assay, which measures the luminescence of luciferase by coupling its expression to the promoter of a gene under study. This luminescence in cells can be measured over time by a luminometer. One major drawback of the luminometer, however, is that it can only measure the luminescence of a group of cells, and cannot follow the differences that may exist among individual cells. A novel luminescence microscope allows the visualization of individual luminescent cells over time through CCD photography. In this study, live single cells of the rat hippocampus were observed under the microscope for luciferase expression driven by the c-fos promoter. We showed that the cell body and neurite areas within a single neuron exhibited differences in luminescence. Because this microscope could detect differences among subcellular regions of single-cell, it may be a promising novel tool to study polarized cells like neurons, and to elucidate proteins involved in neuronal processes such as dendritic/axonal targeting and synaptogenesis.
Asai
Sayaka
S
Mitsubishi Kagaku Institute of Life Sciences (MITILS), 11 Minamiooya, Machida, Tokyo 194-8511, Japan.
Takamura
Kay
K
Suzuki
Hirobumi
H
Setou
Mitsutoshi
M
D000818:Animals; D001369:Axons / Q000378:metabolism / Q000648:ultrastructure; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D006624:Hippocampus / Q000166:cytology* / Q000378:metabolism; D007202:Indicators and Reagents; D008156:Luciferases / Q000737:chemistry / Q000528:radiation effects; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis / Q000378:metabolism; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis* / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D015415:Biomarkers; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D016760:Proto-Oncogene Proteins c-fos; D008156:Luciferases