Physical structure of the excitable membrane of unmyelinated axons: X-ray scattering study and electrophysiological properties of pike olfactory nerve.
The aim of this work was to elicit correlations between physical structure and physiological functions in excitable membranes. Freshly dissected pike olfactory nerves were studied by synchrotron radiation X-ray scattering experiments and their physiological properties were tested by electrophysiological techniques. The scattering spectra contained a sharply oriented equatorial component (i.e. normal to the nerve axis), and an isotropic background. After background subtraction, the equatorial component displayed a weak and fairly sharp spectrum of oriented microtubules, and a strong and diffuse band of almost the same shape and position as the band computed for an isolated myelin membrane. We ascribed this spectrum to the axonal membranes. Under the action of temperature and of two local anesthetics, the spectrum underwent a contraction (or expansion) in the s-direction, equivalent to the structure undergoing an expansion (or contraction) in the direction perpendicular to the plane of the membrane. The main observations were: (i) with increasing temperature, membrane thickness decreased with a thermal expansion coefficient equal to -0.97(+/-0.19) 10(-3) degrees C(-1). The polarity and amplitude of this coefficient are typical of lipid-containing systems with the hydrocarbon chains in a disordered conformation. The amplitude and propagation velocity of the compound action potentials were drastically and reversibly reduced by lowering the temperature from 20 degrees C to 5 degrees C. (ii) Exposing the nerve to two local anesthetics (tetracaine and dibucaine) had the effect of decreasing membrane thickness. Action potentials were fully inhibited by these anesthetics. (iii) Upon depolarization, induced by replacing NaCl with KCl in the outer medium, approximately 25 % of the membranes were found to associate by apposing their outer faces. Electrophysiological activity was reversibly impaired by the KCl treatment. (iv) No detectable structural effect was observed upon exposing the nerves to tetrodotoxin or veratridine. Electrophysiological activity was fully impaired by tetrodotoxin and partially impaired by veratridine. The main conclusions of this work are that axonal membranes yield highly informative X-ray scattering spectra, and that these spectra are sensitive to the functional state of the nerve. These results pave the way to further studies of more direct physiological significance.
Luzzati
V
V
Centre de Génétique Moléculaire, UPR 2167 CNRS associé à l'Université Pierre et Marie Curie, Gif-sur-Yvette Cedex, 91198, France. luzzati@cgm.cnrs-gif.fr
Mateu
L
L
Vachette
P
P
Benoit
E
E
Charpentier
G
G
Kado
R
R
D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D001369:Axons / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology*; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D003992:Dibucaine / Q000494:pharmacology; D004594:Electrophysiology; D017859:Esocidae / Q000502:physiology*; D008870:Microtubules / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D008968:Molecular Conformation; D009186:Myelin Sheath / Q000502:physiology; D009832:Olfactory Nerve / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D011189:Potassium Chloride / Q000494:pharmacology; D013223:Statistics as Topic; D017356:Synchrotrons; D013696:Temperature; D013748:Tetracaine / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D014701:Veratridine / Q000494:pharmacology; D014961:X-Ray Diffraction
D000779:Anesthetics, Local; D013748:Tetracaine; D013779:Tetrodotoxin; D011189:Potassium Chloride; D014701:Veratridine; D003992:Dibucaine