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. 2012 Jun;139(6):493–505. doi: 10.1085/jgp.201110685

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© 2012 Frederiksen et al.

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Figure 3. — Regeneration of visual pigment in isolated salamander rods after extensive bleaching is faster when treated exogenously with 11-cis 4-OH retinal than with 11-cis-retinal. (A) Absorbance spectra measured microspectrophotometrically for comparison of the time course of retinoid partition and pigment regeneration in a bleached salamander rod photoreceptor outer segment after treatment with 11-cis 4-OH retinal. Two peaks appear: one at 385 nm, the retinoid loading into the outer segment, and the other at 478 nm reports the regeneration of the visual pigment. Note that rapid loading of 11-cis 4-OH retinal in the outer segment precedes pigment formation. The numbers in the plot indicate time in minutes after retinal treatment. (B) Absorbance spectra taken after exogenous application of 11-cis-retinal as in A. Note the smaller retinoid peak, compared with A, in the near UV region of the spectrum. The numbers in the panel indicate time in minutes after retinal treatment. (C) Time course of regeneration of visual pigment measured microspectrophotometrically in intact rods treated with 11-cis 4-OH retinal and 11-cis-retinal. Transverse OD in outer segments of intact rods was measured at 520 nm and normalized to avoid bias from retinoid absorbance at shorter wavelength (see A and B). Bleached rods treated with 11-cis-retinal regenerate pigment with a time constant of 12.8 min, n = 6; those treated with 11-cis 4-OH retinal regenerate pigment with a time constant of 2.6 min, n = 5. In both cases, regeneration data are fitted as simple exponential functions. The retinal concentration was 10 µM in 0.1% ethanolic solution. Error bars show means ± SEM. (D) Normalized recovery of sensitivity of bleached rods treated with 11-cis-retinal (open triangles) or 11-cis 4-OH retinal (closed circles). The lines are single exponential functions that were fitted to pigment regeneration data in C. The dashed line represent pigment regeneration with 11-cis-retinal, and the solid line represents 11-cis 4-OH retinal. Note that in both cases, pigment regeneration precedes the recovery of sensitivity.