A novel role for a Drosophila homologue of cGMP-specific phosphodiesterase in the active transport of cGMP

被引:9
作者
Day, JP
Houslay, MD
Davies, SA [1 ]
机构
[1] Univ Glasgow, Inst Biomed & Life Sci, Div Mol Genet, Glasgow G11 6NU, Lanark, Scotland
[2] Univ Glasgow, Div Biochem & Mol Biol, Glasgow G11 6NU, Lanark, Scotland
基金
英国生物技术与生命科学研究理事会; 英国惠康基金; 英国医学研究理事会;
关键词
cGMP; Drosophila; phosphodiesterase 6 (PDE6); renal epithelium; RNA interference (RNAi);
D O I
10.1042/BJ20051505
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
cGMP was first discovered in urine, demonstrating that kidney cells extrude this cyclic nucleotide. Drosophila Malpighian tubules provide a model renal system in which a homologue of mammalian PDE (phosphodiesterase) 6 is expressed. In humans, this cG-PDE (cGMP-specific PDE) is specifically expressed in the retinal system, where it controls visual signal transduction. In order to gain insight into the functional role of DmPDE6 (Drosophila PDE6-like enzyme) in epithelial function, we generated transgenic animals with targeted expression of DmPDE6 to tubule Type I (principal) cells. This revealed localization of DmPDE6 primarily at the apical membranes. As expected, overexpression of DmPDE6 resulted in elevated cG-PDE activity and decreased tubule cGMP content. However, such targeted overexpression of DmPDE6 creates a novel phenotype that manifests itself in inhibition of the active transport and efflux of cGMP by tubules. This effect is specific to DmPDE6 action, as no effect on cGMP transport is observed in tubules from a bovine PDE5 transgenic line which display reduced rates of fluid secretion, an effect not seen in DmPDE6 transgenic animals. Specific ablation of DmPDE6 in tubule principal cells, via expression of a targeted DmPDE6 RNAi (RNA interference) transgene, conferred increased active transport of cGMP, confirming a direct role for DmPDE6 in regulating cGMP transport in tubule principal cells. Pharmacological inhibition of DmPDE6 in wild-type tubules using the cG-PDE inhibitor, zaprinast, similarly results in stimulated cGMP transport. We provide the first demonstration of a novel role for a cG-PDE in modulating cGMP transport and efflux.
引用
收藏
页码:481 / 488
页数:8
相关论文
共 43 条
[1]   Signaling of rat Frizzled-2 through phosphodiesterase and cyclic GMP [J].
Ahumada, A ;
Slusarski, DC ;
Liu, XX ;
Moon, RT ;
Malbon, CC ;
Wang, HY .
SCIENCE, 2002, 298 (5600) :2006-2010
[2]   Genome-wide survey of V-ATPase genes in Drosophila reveals a conserved renal phenotype for lethal alleles [J].
Allan, AK ;
Du, J ;
Davies, SA ;
Dow, JAT .
PHYSIOLOGICAL GENOMICS, 2005, 22 (02) :128-138
[3]   ISOLATION OF ADENOSINE 3,5-MONOPHOSPHATE AND GUANOSINE 3,5-MONOPHOSPHATE FROM RAT URINE [J].
ASHMAN, DF ;
PRICE, TD ;
MELICOW, MM ;
LIPTON, R .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1963, 11 (04) :330-&
[4]  
BRAND AH, 1993, DEVELOPMENT, V118, P401
[5]   Ectopic expression of bovine type 5 phosphodiesterase confers a renal phenotype in Drosophila [J].
Broderick, KE ;
Kean, L ;
Dow, JAT ;
Pyne, NJ ;
Davies, SA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (09) :8159-8168
[6]   Interactions between epithelial nitric oxide signaling and phosphodiesterase activity in Drosophila [J].
Broderick, KE ;
MacPherson, MR ;
Regulski, M ;
Tully, T ;
Dow, JAT ;
Davies, SA .
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 2003, 285 (05) :C1207-C1218
[7]   Transport of cyclic nucleotides and estradiol 17-β-D-glueuronide by multidrug resistance protein 4 -: Resistance to 6-mercaptopurine and 6-thioguanine [J].
Chen, ZS ;
Lee, K ;
Kruh, GD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (36) :33747-33754
[8]   Characteristics of Photoreceptor PDE (PDE6): similarities and differences to PDE5 [J].
Cote, RH .
INTERNATIONAL JOURNAL OF IMPOTENCE RESEARCH, 2004, 16 (Suppl 1) :S28-S33
[9]   Urinary cyclic GMP excretion and blood pressure levels in a general population [J].
Cui, RZ ;
Iso, H ;
Pi, JB ;
Kumagai, Y ;
Yamagishi, K ;
Tanigawa, T ;
Shimojo, N ;
Shimamoto, T .
ATHEROSCLEROSIS, 2004, 172 (01) :161-166
[10]   CAP(2b), a cardioacceleratory peptide, is present in Drosophila and stimulates tubule fluid secretion via cGMP [J].
Davies, SA ;
Huesmann, GR ;
Maddrell, SHP ;
ODonnell, MJ ;
Skaer, NJV ;
Dow, JAT ;
Tublitz, NJ .
AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY, 1995, 269 (06) :R1321-R1326