Expression of PAC 1, an epitope associated with two synapse-enriched glycoproteins and a neuronal cytoskeleton-associated polypeptide in developing forebrain neurons.
The monoclonal antibody PAC 1 (postsynaptic density and cytoskeleton enriched) recognizes an epitope present on two postsynaptic density-enriched glycoproteins of 130,000 (postsynaptic density-enriched glycoprotein 130) and 117,000 mol. wt (postsynaptic density-enriched glycoprotein 117), and a cytoskeleton-enriched polypeptide of 155,000 mol. wt (cp155). The PAC 1 antibody has been used to study the development of the PAC 1 antigens in the developing rat forebrain in vivo and in tissue culture. cp155 is detected by embryonic day 14 and its level continues to rise until the sixth postnatal week. Postsynaptic density-enriched glycoproteins 130 and 117 are also expressed in embryonic brain although the level of postsynaptic density-enriched glycoprotein 130 initially increases more rapidly than that of postsynaptic density-enriched glycoprotein 117. Peak values are observed at postnatal days 4 (postsynaptic density-enriched glycoprotein 117) and 9 (postsynaptic density-enriched glycoprotein 130). The level of post synaptic density-enriched glycoprotein 117 subsequently decreases to some 50% of the peak value by postnatal day 42. Immunocytochemical studies show that PAC 1 immunoreactivity in developing cerebral cortex, detectable by postnatal day 0, is primarily associated with the perikarya and dendrites of pyramidal cells. The immunoreactivity develops as patches of PAC 1-positive neurons, uniform staining of the cortex only being fully established after postnatal day 9. Double-immunofluorescence labelling studies of forebrain cultures prepared from embryonic day 18 animals shows that many, but not all, growth-associated protein 43-positive neurons exhibit PAC 1 immunoreactivity. Some non-neuronal cells also stain with the PAC 1 monoclonal antibody. The growth cones of cultured neurons exhibit PAC 1 immunoreactivity and the PAC 1 antigens are detected on immunodeveloped western blots of isolated growth cones. The PAC 1 epitope is intracellular, but immunoreactivity does not co-localize with F-actin as detected by rhod-amine-phalloidin or with tubulin immunoreactivity. Postsynaptic density-enriched glycoprotein 130 is readily detected on PAC 1 immunodeveloped western blots of forebrain cultures maintained for up to 14 days in vitro. Postsynaptic density-enriched glycoprotein 117 is only poorly expressed by these cultures. The PAC 1 glycoproteins are present in forebrain synaptic membranes and postsynaptic densities at an early stage of development. The synaptic membrane level of postsynaptic density-enriched glycoprotein 130 and postsynaptic density-enriched glycoprotein 117 increases markedly between postnatal days 3 and 8. The level of both glycoproteins detected in postsynaptic densities remain virtually constant from postnatal days 9-90. These results are consistent with functional roles for these molecules in neuronal and synapse development.
Willmott
T
T
Department of Biochemistry, Royal Holloway and Bedford New College, Egham, Surrey, U.K.
Williamson
T L
TL
Mummery
R
R
Hawkes
R B
RB
Can
A
A
Gurd
J W
JW
Gordon-Weeks
P R
PR
Beesley
P W
PW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000096:biosynthesis / Q000276:immunology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000737:chemistry*; D000939:Epitopes / Q000276:immunology*; D015870:Gene Expression; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000276:immunology*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000276:immunology*; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure; D016548:Prosencephalon / Q000166:cytology* / Q000196:embryology / Q000254:growth & development; D051381:Rats; D017208:Rats, Wistar; D013569:Synapses / Q000737:chemistry*; D013570:Synaptic Membranes / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000941:Antigens; D000939:Epitopes; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins