Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
}
TY - JOUR
T1 - Endogenously inhibited protein kinase C in transgenic Drosophila embryonic neuroblasts down regulates the outgrowth of type I and II processes of cultured mature neurons
AU - Broughton, S J
AU - Kane, N S
AU - Arthur, B
AU - Yoder, M
AU - Greenspan, R J
AU - Robichon, A
PY - 1996/3/15
Y1 - 1996/3/15
N2 - 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.
AB - 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.
KW - Amino Acid Sequence
KW - Animals
KW - Animals, Genetically Modified
KW - Calcium-Calmodulin-Dependent Protein Kinases
KW - Cell Aging
KW - Cells, Cultured
KW - Cyclic AMP-Dependent Protein Kinases
KW - Down-Regulation
KW - Drosophila melanogaster
KW - Enzyme Inhibitors
KW - Molecular Sequence Data
KW - Neurons
KW - Phosphorylation
KW - Protein Kinase C
KW - Stem Cells
U2 - 10.1002/(SICI)1097-4644(19960315)60:4<584::AID-JCB14>3.0.CO;2-H
DO - 10.1002/(SICI)1097-4644(19960315)60:4<584::AID-JCB14>3.0.CO;2-H
M3 - Journal article
C2 - 8707897
VL - 60
SP - 584
EP - 599
JO - Journal of Cellular Biochemistry
JF - Journal of Cellular Biochemistry
SN - 0730-2312
IS - 4
ER -