Fluorescence microscopy imaging of a neurotransmitter receptor and its cell membrane lipid milieu
Abstract: Hampered by the diffraction phenomenon, as expressed in 1873 by Abbe, applications of optical microscopy to image biological structures were for a long time limited to resolutions above the ~200 nm barrier and restricted to the observation of stained specimens. The introduction of fluorescence was a game changer, and since its inception it became the gold standard technique in biological microscopy. The plasma membrane is a tenuous envelope of 4 nm–10 nm in thickness surrounding the cell. Because of its highly versatile spectroscopic properties and availability of suitable instrumentation, fluorescence techniques epitomize the current approach to study this delicate structure and its molecular constituents. The wide spectral range covered by fluorescence, intimately linked to the availability of appropriate intrinsic and extrinsic probes, provides the ability to dissect membrane constituents at the molecular scale in the spatial domain. In addition, the time resolution capabilities of fluorescence methods provide complementary high precision for studying the behavior of membrane molecules in the time domain. This review illustrates the value of various fluorescence techniques to extract information on the topography and motion of plasma membrane receptors. To this end I resort to a paradigmatic membrane-bound neurotransmitter receptor, the nicotinic acetylcholine receptor (nAChR). The structural and dynamic picture emerging from studies of this prototypic pentameric ligand-gated ion channel can be extrapolated not only to other members of this superfamily of ion channels but to other membrane-bound proteins. I also briefly discuss the various emerging techniques in the field of biomembrane labeling with new organic chemistry strategies oriented to applications in fluorescence nanoscopy, the form of fluorescence microscopy that is expanding the depth and scope of interrogation of membrane-associated phenomena.
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Format: | Artículo biblioteca |
Language: | eng |
Published: |
Frontiers Media
2022
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Subjects: | MEMBRANA PLASMATICA, INTERACCIONES PROTEÍNA-PROTEÍNA, COLESTEROL, NANOSCOPIA, MICROSCOPIO FLUORESCENTE, RECEPTOR DE ACETILCOLINA, |
Online Access: | https://repositorio.uca.edu.ar/handle/123456789/18149 |
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Summary: | Abstract: Hampered by the diffraction phenomenon, as expressed in 1873 by Abbe,
applications of optical microscopy to image biological structures were for a
long time limited to resolutions above the ~200 nm barrier and restricted to the
observation of stained specimens. The introduction of fluorescence was a game
changer, and since its inception it became the gold standard technique in
biological microscopy. The plasma membrane is a tenuous envelope of
4 nm–10 nm in thickness surrounding the cell. Because of its highly versatile
spectroscopic properties and availability of suitable instrumentation,
fluorescence techniques epitomize the current approach to study this
delicate structure and its molecular constituents. The wide spectral range
covered by fluorescence, intimately linked to the availability of appropriate
intrinsic and extrinsic probes, provides the ability to dissect membrane
constituents at the molecular scale in the spatial domain. In addition, the
time resolution capabilities of fluorescence methods provide complementary
high precision for studying the behavior of membrane molecules in the time
domain. This review illustrates the value of various fluorescence techniques to
extract information on the topography and motion of plasma membrane
receptors. To this end I resort to a paradigmatic membrane-bound
neurotransmitter receptor, the nicotinic acetylcholine receptor (nAChR). The
structural and dynamic picture emerging from studies of this prototypic
pentameric ligand-gated ion channel can be extrapolated not only to other
members of this superfamily of ion channels but to other membrane-bound
proteins. I also briefly discuss the various emerging techniques in the field of
biomembrane labeling with new organic chemistry strategies oriented to
applications in fluorescence nanoscopy, the form of fluorescence
microscopy that is expanding the depth and scope of interrogation of
membrane-associated phenomena. |
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