Angiotensin type 2 receptor neuroprotection against chemical hypoxia is dependent on the delayed rectifier K+ channel, Na+/Ca2+ exchanger and Na+/K+ ATPase in primary cortical cultures.
We have previously reported that angiotensin II (Ang II) protects cortical neurons from chemical-induced hypoxia through activation of the angiotensin type 2 (AT(2)) receptor. Here, we show in mouse primary neuronal cultures that the AT(2) receptor neuroprotection results from the activation of the delayed rectifier K(+) channel as well as the involvement of the Na(+)/Ca(2+) exchanger (NCX) and Na(+)/K(+) ATPase (ATPase). Roles of the K(+) channel, NCX and ATPase were determined using the specific blockers alpha-dendrotoxin, KB-R7943 and ouabain, respectively. Sodium azide (10mM) induced apoptosis in 40% of neurons. Inhibition of the AT(1) receptor with losartan (1 microM) facilitated angiotensin II mediated neuroprotection by reducing sodium azide-induced apoptosis 61.8 +/- 5.6%, while inhibition of the AT(2) receptor with PD123319 (1 microM) showed no neuroprotection. These results suggest that angiotensin II neuroprotection is mediated through the AT(2) receptor and requires inhibition of the AT(1) receptor in order to facilitate its effect. To determine the roles of delayed rectifier K(+) channel, NCX and ATPase cultures were pretreated with alpha-dendrotoxin (10nM), KB-R7943 (100 nM) and ouabain (100 nM), which significantly attenuated AT(2) receptor mediated neuroprotection. These findings further suggest that the mechanism of AT(2) receptor mediated neuroprotection is coupled to activation of the delayed rectifier K(+) channel, NCX and ATPase.
Department of Cell and Structural Biology, University of Illinois, 346 Henry Administration Building, 506 South Wright Street, Urbana, IL 61801, USA.
Tom N
TN
Grammatopoulos
Victoria
V
Johnson
Steve A
SA
Moore
Robert
R
Andres
James A
JA
Weyhenmeyer
D000818:Animals; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism; D051668:Delayed Rectifier Potassium Channels; D004305:Dose-Response Relationship, Drug; D051379:Mice; D008813:Mice, Inbred ICR; D018696:Neuroprotective Agents / Q000378:metabolism; D026902:Potassium Channel Blockers / Q000494:pharmacology; D024642:Potassium Channels, Voltage-Gated / Q000037:antagonists & inhibitors / Q000378:metabolism*; D044139:Receptor, Angiotensin, Type 2 / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D051668:Delayed Rectifier Potassium Channels; D018696:Neuroprotective Agents; D026902:Potassium Channel Blockers; D024642:Potassium Channels, Voltage-Gated; D044139:Receptor, Angiotensin, Type 2; D019831:Sodium-Calcium Exchanger; D000254:Sodium-Potassium-Exchanging ATPase