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M. Haeri, A. Adekeye, S. Baker, V. Arshavsky, B. Knox; Visualization of Different Organelles and Compartments in Photoreceptors. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1275. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To study the localization of selected proteins and to visualize major organelles and cellular compartments in photoreceptors using live cell imaging, we targeted fluorescent proteins to these locations via specific amino acid sequences.
We prepared transgenic Xenopus tadpoles that expressed tagged fluorescent proteins in photoreceptors using the Xenopus rhodopsin promoter. Live and fixed retinas were examined by high resolution confocal imaging to determine the localization of fluorescent proteins.
We successfully targeted fluorescent protein to the photoreceptor endoplasmic reticulum and confirmed its localization by immunostaining. We also targeted Golgi apparatus, outer segment, mitochondria and nucleus. Using high definition confocal microscopy on live photoreceptors, we generated three dimensional images of above mentioned organelles and compartments.
The 3D images of photoreceptors at subcellular resolution demonstrate detailed features of cell components which can be used for colocalization studies in live photoreceptor imaging.
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