title: Does membrane fluidity contribute to thermal compensation of beta-adrenergic signal transduction in isolated trout hepatocytes?
abstract: The potential role of compensatory adjustments to membrane components in determining the function of the (&bgr;)-adrenergic receptor/adenylyl cyclase ((&bgr;)-AR/AC) signal-transduction system was studied in isolated hepatocytes of 5 degrees C- and 20 degrees C-acclimated rainbow trout Oncorhynchus mykiss. Rates of epinephrine-stimulated cyclic AMP (cAMP) production, although slowed (by a factor of 1.6- to 2.4-fold) by an acute drop in assay temperature from 20 to 5 degrees C, were significantly temperature-compensated, being approximately twofold higher in hepatocytes of 5 degrees C- than of 20 degrees C-acclimated trout. Membrane order in the bilayer interior of hepatocyte plasma membranes (as assessed by fluorescence polarization of 1, 6-diphenyl-1,3,5-hexatriene) was consistently lower in cold- than in warm-acclimated trout, reflecting an efficacy of homeoviscous adaptation of approximately 50 %. Temperature-induced changes in plasma membrane fatty acid composition (i.e. an increase in the proportions of polyunsaturated fatty acids with acclimation to 5 degrees C) were consistent with both the observed changes in the order of the bilayer interior and the extent of homeoviscous adaptation. However, isothermal fluidization of the bilayer interior by the addition of benzyl alcohol (30 mmol l(-)(1)) decreased rather than increased the rate of cAMP production. Significantly more (1. 81-fold) beta-adrenergic receptors were present in plasma membranes of hepatocytes from 5 degrees C-acclimated (6. 23x10(4)+/-4206 receptors per cell; mean +/- s.e.m., N=3) than 20 degrees C-acclimated fish (3.44x10(4)+/-4360 receptors per cell; N=3) when assayed at the acclimation temperature, whereas the equilibrium dissociation constants (K(d)) (13.73+/-4.33 nmol l(-)(1) at 5 degrees C; 9.75+/-3.29 nmol l(-)(1) at 20 degrees C; N=3) were similar. On the basis of a strong correlation between beta-adrenoceptor number and the rate of cAMP production (r(2)=0. 956), regardless of assay or acclimation temperature, we conclude that modulation of receptor number is the primary acclimatory response of this signal-transduction pathway to temperature change in trout liver.
authors.affiliation: Department of Biology, Arizona State University, Tempe, AZ 85287-1501, USA. J.hazel@asu.edu
authors.forename: S J
authors.initials: SJ
authors.lastname: McKinley
authors.forename: J R
authors.initials: JR
authors.lastname: Hazel
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000322:Adrenergic Agonists / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D000242:Cyclic AMP / Q000096:biosynthesis; D004837:Epinephrine / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000737:chemistry; D005260:Female; D005454:Fluorescence Polarization; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology / Q000502:physiology; D008297:Male; D008560:Membrane Fluidity / Q000502:physiology*; D017686:Oncorhynchus mykiss / Q000502:physiology*; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013696:Temperature
chemical_list: D000322:Adrenergic Agonists; D005231:Fatty Acids, Unsaturated; D011943:Receptors, Adrenergic, beta; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D004837:Epinephrine