Effect of electric fields on light-scattering and fluorescence of chromaffin granules

Abstract

Electric field pulses (in the 5 to 25 kV/cm range) were found to cause a transient increase in the intensity of scattered light (wavelength equals 369 nm, scattering angle equals 90 degrees) from aqueous sucrose suspensions of chromaffin granules. Similar observations were made with the membranes of osmotically lysed chromaffin granules. Under the same experimental conditions the degree of polarization of the scattered light changed only very slightly. The fluorescence of the hydrophobic probe diphenyl-hexatriene, incorporated into the membrane of intact chromaffin granules, showed similar transient changes in the intensity. The calculated relaxation times for these changes in optical properties were approximately 150 musec for the rising phase, and approximately 1 msec for the early stage of the decay. A further relaxation time of about 30 msec was also observed by using this probe. Essentially, all of these signals originated from the granule membrane, and could be attributed to rather small changes in particle size, membrane thickness or refractive index. Moreover, these signals were found to be completely reversible. Catecholamine release from intact granules, pulsed at voltages of 25 kV/cm, occurs already during the first few milliseconds of the transient membrane change.