Skip to main content
construction release_alert
Scholars@Duke will be undergoing maintenance April 11-15. Some features may be unavailable during this time.
cancel

Elevation of cAMP is required for down-regulation, but not agonist-induced desensitization, of endogenous dopamine D1 receptors in opossum kidney cells. Studies in cells that stably express a rat cAMP phosphodiesterase (rPDE3) cDNA.

Publication ,  Journal Article
Bates, MD; Olsen, CL; Becker, BN; Albers, FJ; Middleton, JP; Mulheron, JG; Jin, SL; Conti, M; Raymond, JR
Published in: J Biol Chem
July 15, 1993

D1 dopamine receptors stimulate cAMP accumulation in opossum kidney (OK) cells, but this response is attenuated by pretreatment with dopamine. Dopamine pretreatment also causes a reduction in D1 dopamine receptor number. We transfected OK cells with a rat cAMP phosphodiesterase cDNA (rPDE3) in order to determine the contribution of elevations of cAMP to those two phenomena. Wild-type (WT) OK cells were compared to three clones (C, H, and N) which demonstrated stable expression of the rPDE3 phenotype and genotype, rPDE3 RNA expression was confirmed in clones C, H, and N (but not in WT-OK cells) by reverse transcriptase-polymerase chain reaction. A functional rPDE3 phenotype was demonstrated in that dopamine-responsive cAMP accumulation was absent in clones C, H, and N in intact cells, but could be restored by preincubation with cAMP phosphodiesterase inhibitors, or by using washed membranes from those clones. All three clones had increased cAMP phosphodiesterase activity when compared to WT-OK cells (approximately 100% increase), and blunted or absent dopamine (1 microM)-induced protein kinase A activation. After pretreatment with dopamine (1 microM) for 1 h, clones C, H, and N desensitized equally well as WT-OK cells (approximately 40-50% reduction in maximal increase in cAMP). In contrast, down-regulation of D1 dopamine receptors was blunted for clone C (20% receptor loss) and absent for clones H and N, when compared to a 45% loss of receptors for WT-OK cells. These findings suggest that in OK cells pretreated with 1 microM dopamine (i) cAMP accumulation is not necessary for dopamine-induced desensitization, but (ii) is necessary for down-regulation of D1 dopamine receptors, and (iii) that the down-regulation and desensitization processes may be differentially regulated.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

July 15, 1993

Volume

268

Issue

20

Start / End Page

14757 / 14763

Location

United States

Related Subject Headings

  • Receptors, Dopamine D1
  • Rats
  • Protein Kinases
  • Polymerase Chain Reaction
  • Opossums
  • Kidney
  • Enzyme Activation
  • Down-Regulation
  • Dopamine
  • Depression, Chemical
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Bates, M. D., Olsen, C. L., Becker, B. N., Albers, F. J., Middleton, J. P., Mulheron, J. G., … Raymond, J. R. (1993). Elevation of cAMP is required for down-regulation, but not agonist-induced desensitization, of endogenous dopamine D1 receptors in opossum kidney cells. Studies in cells that stably express a rat cAMP phosphodiesterase (rPDE3) cDNA. J Biol Chem, 268(20), 14757–14763.
Bates, M. D., C. L. Olsen, B. N. Becker, F. J. Albers, J. P. Middleton, J. G. Mulheron, S. L. Jin, M. Conti, and J. R. Raymond. “Elevation of cAMP is required for down-regulation, but not agonist-induced desensitization, of endogenous dopamine D1 receptors in opossum kidney cells. Studies in cells that stably express a rat cAMP phosphodiesterase (rPDE3) cDNA.J Biol Chem 268, no. 20 (July 15, 1993): 14757–63.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

July 15, 1993

Volume

268

Issue

20

Start / End Page

14757 / 14763

Location

United States

Related Subject Headings

  • Receptors, Dopamine D1
  • Rats
  • Protein Kinases
  • Polymerase Chain Reaction
  • Opossums
  • Kidney
  • Enzyme Activation
  • Down-Regulation
  • Dopamine
  • Depression, Chemical