PHOTOSENSORY TRANSDUCTION IN CILIATES. II. POSSIBLE ROLE OF G‐PROTEIN AND cGMP IN <i>Stentor coeruleus</i>

Abstract

The heterotrichous ciliate, Stentor coeruleus, exhibits a well-defined photophobic response to a sudden increase in the intensity of visible light. The phobic reactions usually appear with a latency period (i.e. a time delay between the onset of the stimulus and the stop response). This latency of phobic response was significantly increased when the cells were incubated with 8-bromo-guanosine 3',5'-cyclic monophosphate. In the presence of this nucleotide, a reduction of cell responsiveness (i.e. the number of photophobically responding cells) was also observed. Similar effects were observed when cells were treated with pertussis toxin, a G-protein activity modulator, and 3'-isobutyl-methylxanthine, an inhibitor of guanosine 3',5'-cyclic monophosphate (cGMP) phosphodiesterase. The G-protein activator fluoroaluminate and 6-anilino-5,8-quinolinedione (LY 83583) (an effective agent for lowering cellular cGMP levels) showed opposite effects on the cell photophobic response. These results indirectly suggest that the level of cytoplasmic cGMP, possibly modulated by a G-protein-coupled cGMP phosphodiesterase, plays a phototransducing role in Stentor. In addition, using an antiserum raised against bovine transducin, a cross-reacting protein with an apparent molecular mass of 39 kDa was detected on immunoblots. The alpha-subunit of a Stentor G-protein has also been partially cloned and sequenced. However, the possible coupling between the G-protein and the putative phosphodiesterase remains to be established.