The effect of the angiotensin II (AT1A) receptor stably transfected into human neuroblastoma SH-SY5Y cells on noradrenaline release and changes in intracellular calcium

Neurosci Lett. 1995 Oct 20;199(2):115-8. doi: 10.1016/0304-3940(95)12035-3.

Abstract

A stable cell line expressing the angiotensin II (AII) receptor has been obtained by transfecting the human neuroblastoma SH-SY5Y with the plasmid pCEP4 containing the entire coding region of the rat angiotensin AII receptor AT1A. Angiotensin II (AII; 1-100 nM) evokes the release of [3H]noradrenaline ([3H]NA) in this cell line. Pretreatment with 100 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) enhances the AII-evoked release of [3H]NA approximately two-fold. Removal of extracellular Ca2+ ([Ca2+]o) decreases 100 nM AII-evoked release of [3H]NA by over 50% both in the presence and absence of TPA. AII increases intracellular Ca2+ ([Ca2+]i) in this cell line which is consistent with the AT1A receptor being coupled to phospholipase C. Pretreatment with 100 nM TPA for 8 min attenuated the effect of AII on [Ca2+]i. The effects of AT1A receptor stimulation are therefore regulated differently in this cell line by activation of protein kinase C (PKC). Thus a useful cell line has been obtained from the human neuroblastoma SH-SY5Y in which to study at the molecular level the mechanism(s) by which AII regulates NA release.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin II / metabolism*
  • Angiotensin II / pharmacology
  • Animals
  • Brain Neoplasms / metabolism*
  • Calcium / metabolism*
  • Calcium / physiology
  • Humans
  • Neuroblastoma / metabolism*
  • Norepinephrine / metabolism*
  • Protein Kinase C / metabolism
  • Rats
  • Receptors, Angiotensin / genetics
  • Receptors, Angiotensin / metabolism*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Receptors, Angiotensin
  • Angiotensin II
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate
  • Calcium
  • Norepinephrine