. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 7 Figure 3. PCR amplification of genomic DNA using Haliotis-sped&c pnmers S, and S: (30 cycles). Lanes: 11) molecular weight standards; (2) and (3) Mg Haliolis sperm DNA; (4) ^g Tetrahymena DNA; (5) Mg Vibrio DNA; (6) 1 Mg salmon sperm DNA; (7) no DNA. Other details in Materials and Methods. from the environment (Morse, 1990a, b). Purification of cilia in milligram quantities from the cultured larvae has allowed us to analyze the chemosensory receptors and signal transducers /// vitro (Baxter, 1991; Baxter a
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 7 Figure 3. PCR amplification of genomic DNA using Haliotis-sped&c pnmers S, and S: (30 cycles). Lanes: 11) molecular weight standards; (2) and (3) Mg Haliolis sperm DNA; (4) ^g Tetrahymena DNA; (5) Mg Vibrio DNA; (6) 1 Mg salmon sperm DNA; (7) no DNA. Other details in Materials and Methods. from the environment (Morse, 1990a, b). Purification of cilia in milligram quantities from the cultured larvae has allowed us to analyze the chemosensory receptors and signal transducers /// vitro (Baxter, 1991; Baxter and Morse, in prep.), and (as shown here) to isolate mRNAs encoding some of these elements and conduct analyses at the cDNA sequence level. We have shown here that cilia purified from //. rufescens larvae contain polyadenylated mRNA, and that this mRNA includes sequences corre- sponding to two Ga signal transducing proteins. The central role of the G proteins as chemosensory signal transducers was confirmed by /// vitro studies of olfactory cilia isolated from frog (Pace et ai, 1985). Jones and Reed (1989) recently have identified a unique G0|f from the ciliated sensory epithelium of the rat olfactory mucosa; the sequence of this G protein a subunit was determined by analysis of its cDNA. G protein also acts as the primary chemosensory signal transducer in taste receptor cells in the frog; this G protein controls the ac- tivation of adenyl cylase, with the resulting sequential ac- tivation of a protein kinase and membrane depolarization (Avenet et ai, 1988). G protein transduction of chemo- sensory stimulation in catfish controls an inositol tri- phosphate (protein kinase) cascade (Huque et ai. 1987). Although distal localization of mRNA has been ob- served in other systems (Merlie and Sanes, 1985; Garner et ai. 1988; Kosik et ai. 1989), the results reported here are the first of which we are aware in which mRNA has been purified, and the corresponding cDNA synthesized, amplifie
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