Phospholipase Cgamma1 in bovine rod outer segments: immunolocalization and light-dependent binding to membranes.
We have investigated the isozymes of a phosphoinositide-specific phospholipase C (PLC) in bovine retina using several monoclonal antisera to PLCbeta1, gamma1, and delta1. Immunoblot analysis showed that all three isozymes were present in the retina. Immunocytochemical localization in frozen bovine retina sections showed that PLCgamma1 was present in the photoreceptor cell layer, outer plexiform cell layer, inner plexiform cell layer, and ganglion cell layer. Immunoreaction within the photoreceptor cell layer was dependent on dark/light adaptation state of retinas. Immunoblot analysis of rod outer segments (ROS) with monoclonal or polyclonal antibodies to PLCgamma1 showed the presence of an immunoreactive band of 140 kDa. ROS prepared from retinas light-adapted in vitro had more PLCgamma1 on immunoblots than ROS from dark-adapted retinas. PLC enzyme activity in ROS from light-adapted retinas was 69 and 46% higher than ROS from dark-adapted retinas, when assayed in the presence and absence of ATP, respectively. This increase in enzyme activity was observed at [Ca2+]free between 0.32 and 100 microM. These results demonstrate the presence of PLCgamma1 in bovine ROS and show that ROS prepared from light-adapted retinas are enriched in this isozyme, suggesting that light may promote the binding of this isozyme to bleached ROS membranes.
Ghalayini
A J
AJ
Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City 73104, USA.
Weber
N R
NR
Rundle
D R
DR
Koutz
C A
CA
Lambert
D
D
Guo
X X
XX
Anderson
R E
RE
D000221:Adaptation, Ocular / Q000502:physiology; D000818:Animals; D002417:Cattle; D003623:Dark Adaptation / Q000502:physiology; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D008027:Light; D008566:Membranes / Q000201:enzymology / Q000528:radiation effects; D051966:Phospholipase C gamma; D011233:Precipitin Tests; D012160:Retina / Q000201:enzymology; D012374:Rod Cell Outer Segment / Q000201:enzymology* / Q000528:radiation effects*; D014018:Tissue Distribution; D010738:Type C Phospholipases / Q000378:metabolism*
D007527:Isoenzymes; D010738:Type C Phospholipases; D051966:Phospholipase C gamma