Measurements of fast light-induced light-scattering and -absorption changes in outer segments of vertebrate light sensitive rod cells

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Abstract

Flash-induced changes of light-absorption and of light-scattering of vertebrate rod outer segments (ROS) from frog and cattle in suspension were measured at 380 and 800 nm. The photometer used allows the observation of light intensity changes under well defined angles.

We studied the successive decrease of the signal amplitude in series of flashes. One flash bleaches about 1% rhodopsin.

The following results are discussed:

  1. The signal at 380 nm is a superposition of the absorption change caused by formation of metarhodopsin II and of a biphasic additional signal. The latter exists only for the initial range of bleaching (15 to 25% rhodopsin).

  2. At 800 nm three scattering signals are observed which are characterized by their successive amplitude decrease and time course:

N: A small signal with time course and successive amplitude decrease comparable to the metarhodopsin II absorption change, probably arising from a structural change within the disc membrane.

Ni: A slow signal, disappearing with the first flash, which may be understood as an outer membrane effect.

P: A biphasic signal with a successive decrease rate, by a factor of 10 to 20 higher than that of the metarhodopsin II signal. The two kinetically different components are separated by variation of the observation angle. Two regions of different extension appear to change structurally with different time course. “P” may reflect an influence of the light-induced transmitter release on disc shape and/or mass.

Presented in part at the Annual Meeting of the Deutsche Gesellschaft für Biophysik, Freiburg, October 1974, and at the Membrane Symposium, Freiburg, April 1975; in connection with new results at the International Biophysics Congress, Copenhagen, August 1975.