Endogenously inhibited protein kinase C in transgenic Drosophila embryonic neuroblasts down regulates the outgrowth of type I and II processes of cultured mature neurons.
Embryonic neurons were cultured from transgenic Drosophila melanogaster expressing a highly specific pseudosubstrate inhibitor of protein kinase C (PKC). Flies homozygous for this transgene, which is under the control of the yeast UAS promoter, were crossed to flies homozygous for the yeast heat shock inducible transcription factor GAL 4. Following heat shock, the progeny express the pseudosubstrate inhibitor at high levels. This strategy, which has the advantage of avoiding the non-specific effects of drugs, was used to study the role of PKC in process growth of cultured, differentiating neuroblasts. An external gold particle labeling procedure using a cell surface antigen expressed by mature neurons and processes was used to visualize neuronal processes directly in the scanning electron microscope. We observed that cell cultures expressing a low concentration of the pseudosubstrate inhibitor showed a significant decrease in the number of type I and II processes as compared to control cultures, while the proportions of neuroblasts, ganglion mother cells (GMCs), and mature neurons in the clusters were little affected.
Broughton
S J
SJ
Department of Biology, New York University, New York 10003, USA.
Kane
N S
NS
Arthur
B
B
Yoder
M
M
Greenspan
R J
RJ
Robichon
A
A
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D015536:Down-Regulation; D004331:Drosophila melanogaster / Q000196:embryology / Q000201:enzymology / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D008969:Molecular Sequence Data; D009474:Neurons / Q000201:enzymology* / Q000648:ultrastructure; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D013234:Stem Cells / Q000201:enzymology*
D004791:Enzyme Inhibitors; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases