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Phospholipase Cγ 1 in Bovine Rod Outer Segments: Immunolocalization and Light‐Dependent Binding to Membranes
Author(s) -
Ghalayini Abboud J.,
Weber Nathan R.,
Rundle Dana R.,
Koutz Cynthia A.,
Lambert David,
Guo Xiao X.,
Anderson Robert E.
Publication year - 1998
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1998.70010171.x
Subject(s) - phospholipase c , ganglion cell layer , retina , biology , outer plexiform layer , microbiology and biotechnology , inner plexiform layer , isozyme , polyclonal antibodies , biochemistry , enzyme , signal transduction , antibody , immunology , neuroscience
We have investigated the isozymes of a phosphoinositide‐specific phospholipase C (PLC) in bovine retina using several monoclonal antisera to PLCβ 1 , γ 1 , and δ 1 . Immunoblot analysis showed that all three isozymes were present in the retina. Immunocytochemical localization in frozen bovine retina sections showed that PLCγ 1 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 PLCγ 1 showed the presence of an immunoreactive band of 140 kDa. ROS prepared from retinas light‐adapted in vitro had more PLCγ 1 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 [Ca 2+ ] free between 0.32 and 100 µ M . These results demonstrate the presence of PLCγ 1 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.