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Preparation of Living Isolated Vertebrate Photoreceptor Cells for Fluorescence Imaging
Published on: June 22, 2011
Temperature effects on spectral properties of red and green rods in toad retina
Petri Ala-Laurila1, Pia Saarinen, Rauli Albert
1Laboratory of Biomedical Engineering, Helsinki University of Technology, FIN-02015 HUT, Finland. Petri.Ala-Laurila@hut.fi
Abstract:
Temperature effects on spectral properties of the two types of rod photoreceptors in toad retina, "red" and "green" rods, were studied in the range 0-38 degrees C. Absorbance spectra of the visual pigments were recorded by single-cell microspectrophotometry (MSP) and spectral sensitivities of red rods were measured by electroretinogram (ERG) recording across the isolated retina. The red-rod visual pigment is a usual rhodopsin (lambda(max) = 503.6 nm and 502.3 nm at room temperature (21degrees C) in, respectively, Bufo marinus and Bufo bufo), that of green rods (lambda(max) = 432.6 nm in Bufo marinus) belongs to the "blue" cone pigment family. In red rods, lambda(max) depended inversely and monotonically on temperature, shifting by -2.3 nm when temperature was raised from 0 degrees C to 38 degrees C. Green-rod lambda(max) showed no measurable dependence on temperature. In red rods, warming caused a relative increase of sensitivity in the long-wavelength range. This effect can be used for estimating the energy needed for photoexcitation, giving Ea = 44.3 +/- 0.6 kcal/mol for Bufo marinus rhodopsin and 48.8 +/- 0.5 kcal/mol for Bufo bufo rhodopsin. The values are significantly different (P < 0.001), although the two rhodopsins have very similar absorption spectra and thermal isomerization rates. Our recording techniques did not allow measurement of the corresponding effect at long wavelengths in green rods. Although spectral effects of temperature changes in the physiological range are small and of little significance for visual function, they reveal information about the energy states and different spectral tuning mechanisms of the visual pigments.
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