Differential Expression and Subcellular Localization of Protein Kinase C α, β, γ, δ, and ɛ Isoforms in SH‐SY5Y Neuroblastoma Cells: Modifications During Differentiation

Leli, Ubaldo; Shea, Thomas B.; Cataldo, Anne M.; Hauser, George; Grynspan, Frida; Beermann, Mary Lou; Liepkalns, Vis A.; Nixon, Ralph A.; Parker, Peter J.

doi:10.1111/j.1471-4159.1993.tb05850.x

Cited by 69 publications

(33 citation statements)

References 56 publications

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“…2). In contrast to a previous observation on another NB cell line [23] we did not find any PKC /I mRNA. This could be ascribed to differences in method evaluation or to a further cell clone-specificity.…”

Section: Resultscontrasting

confidence: 57%

“…Previous reports have documented changes in PKC during neuronal maturation [22][23][24]331. While these studies have demonstrated modulation of some PKC isoenzymes during treatment of neural cells with PKC inhibitors or they have focused on only few PKC isoforms, they have, however, failed to address whether true differentiation-promoting agents, such as IFN-)I and RA, affect the individual isoforms in a differential manner within the same neuronal model.…”

Section: Resultsmentioning

confidence: 99%

“…Several proteins which participate in neurofilament reorganization during neural differentiation are themselves substrates for PKC [37]. Moreover, PKCE is nearly exclusively localized in the membranes and processes in NB cells [23]. Thus, it is possible that both the PKCE isoform and particular substrates colocalize during neurite extension and that proximal distribution of the kinase and substrate plays a key role in the acquisition of neuronal phenotype.…”

Section: Febs Letters May 1993mentioning

confidence: 99%

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Protein kinase C isoenzymes in human neuroblasts involvement of PKCε in cell differentiation

et al. 1993

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Although neuronal cells are a major target of phorbol ester action, the activity of the various protein kinase C (PKC) isoenzymes have not been studied in detail in human neuroblasts. Differentiation of the LAN-5 human neuroblastoma cell line by interferon-y (IFN-y) is accompanied by a twofold increase in PKC activity. Since PKC is a multigene family, we investigated which isoforms were expressed in control and differentiated cells, and which of these isoenzymes is involved in neuronal differentiation. We found that: (1) PKC activity is higher in differentiated than in undifferentiated cells; (2) RT-PCR analysis showed the expression of mRNA for PKCa, -y, -6, -E and -c and the absence of mRNA forp in untreated LAN-5 cells; (3) Western blot evaluation with PKC isoform-specific antibodies showed the same pattern of PKC expression in non-differentiated cells; (4) Expression of PKCE mRNA was significantly enhanced by IFN-y-induced differentiation, while the other isoforms were not affected; (5) Differentiation of LAN-5 cells with IFN-I or retinoic acid induced overexpression of the PKCE protein, while inhibition of cell proliferation by fetal calf serum starvation was without effect. These findings suggest that expression of PKCE isoform is tightly coupled with neuronal differentiation and may play a role in the maintenance of the differentiated state.

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Section: Resultscontrasting

confidence: 57%

Section: Resultsmentioning

confidence: 99%

Section: Febs Letters May 1993mentioning

confidence: 99%

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Protein kinase C isoenzymes in human neuroblasts involvement of PKCε in cell differentiation

et al. 1993

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“…This is a difference between NG 108-15 cells and other neuroblastoma cells used to study the PKC isoforms in the context of differentiation. The human neuroblastoma cell line SH-SYSY was shown to express PKC a, P, y, 6 and E isoforms, and the PKC a and the PKC E isoforms are downregulated during differentiation (Leli et al, 1993). The same behavior was observed with the murine neuroblastoma cell line Neuro 2a, where selectively PKC a and PKC E are downregulated (Wada et al, 1989).…”

Section: Pkc Isozymes In Subcellular Fractionssupporting

confidence: 49%

Differential nuclear localization of protein kinase C isoforms in neuroblastoma × glioma hybrid cells

Beckmann

Lindschau

Haller

et al. 1994

European Journal of Biochemistry

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The protein kinase C (PKC) a, p and E isoforms have distinct nuclear localizations in neuroblastoma X glioma hybrid cells NG 108-15. We found by immunoblotting that PKC a, pII, 6 and E are the predominant isoforms in these cells. In contrast to other neuronal cell lines, none of these isoforms is down-regulated during differentiation.Confocal immunofluorescence microscopy revealed that in undifferentiated cells PKC a is located in the cytoplasm and in the nucleus excluding nucleoli. In differentiated cells PKC a was almost exclusively located in the cytoplasm. Stimulation of the cells with phorbol ester resulted in translocation to the plasma membrane. PKC PII was not detectable in the nuclei. PKC 6 was found in the nucleoli and in the cytoplasm, in differentiated cells particularly in the neurites. Phorbol ester failed to induce a translocation to other compartments. PKC E was localized with the nuclear-pore complexes at the nuclear envelope. In differentiated cells after stimulation with phorbol ester, partial translocation to the plasma membrane was observed.Protein kinase C (PKC) plays a key role in many signaltransduction pathways and cellular processes (for review see Nishizuka, 1986;Lester and Epand, 1992). Ten isoforms of PKC are found in mammals, showing characteristic patterns of expression in different tissues. Four isoforms (called PKC A forms), PKC a, PI, pII and 7, are phospholipid and Ca2+ dependent in contrast to the B forms such as PKC 6, E and cwhich are Ca2+ independent (Stabel and Parker, 1991). After an increase of intracellular Ca2+ levels the PKC A isoforms were shown to be translocated to the plasma membrane. Binding to the plasma membrane via phospholipids enables activation by the second messenger diacylglycerol (Huang, 1989).There is evidence that members of the PKC family respond differently to various combinations of Caz+ and phospholipid-degradation products. This could be one of the reasons for distinct intracellular distribution and activation patterns (Asaoka et al., 1992).Several investigators have observed translocation to other cell compartments including the cell nucleus in addition to the plasma membrane. With fibroblast cells, Leach et al. (1992) observed translocation of PKC a to the cell nucleus after treatment with phorbol ester or a-thrombin. In promyelocytic leukemia (HL 60) cells, stimulation with the PKC activator bryostatin leads to a selective translocation of PKC P but not PKC a to the nucleus (Hocevar and Fields, 1991).In liver, PKC BI1 was found in the nuclei (Rogue et al., 1990 found PKC a and PKC y as the 'membrane inserted' and constitutively active form.The current view is that each cell type expresses a specific set of PKC isoforms, which are localized and regulated in a cell-specific manner. It is not yet understood how this differential expression, localization, and regulation is achieved. The specific localizations of PKC isoforms imply specific functions for each cell type. Due to the known effects of PKC activation on nuclear events (induction of proli...

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“…However, despite the efforts that have been made, no clear agreement has been reached onto whether an increase or a decrease of PKC activity is associated to neuronal differentiation. Down-regulation or inhibition of PKC activity has been associated to initiation of neuronal differentiation in N2A neuroblastoma cells [37] and SH-SY5Y neuroblastoma cells [38][39][40]. Also, dibutyryl cAMP-induced differentiation of NG108-15 cells is accompanied by a decrease in PKC~z [24].…”

Section: Phase Contrastmentioning

confidence: 99%

Expression of protein kinase C isozymes in hippocampal neurones in culture

1995

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Several protein kinase C (PKC) isozymes were analyzed by immunoblot and immunocytochemistry in cultures of hippocampal neurones at several stages of differentiation. Our findings reveal the existence of two distinct patterns of expression. Firstly, conventional PKC isozymes a, /3 and % that are expressed at very low levels during the initial stages and then increase continuously with time of culture. Secondly, novel PKC isozymes 6, ~ and 5, whose contents increase very early to reach a maximum after three days of culture and then progressively decline. Specific proteolysis for PKC isozymes i~ and y was observed throughout the period studied. The developmental profile obtained for the different PKC isozymes is discussed in relation to the differentiation of hippocampal neurones in culture.

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Differential Expression and Subcellular Localization of Protein Kinase C α, β, γ, δ, and ɛ Isoforms in SH‐SY5Y Neuroblastoma Cells: Modifications During Differentiation

Cited by 69 publications

References 56 publications

Protein kinase C isoenzymes in human neuroblasts involvement of PKCε in cell differentiation

Protein kinase C isoenzymes in human neuroblasts involvement of PKCε in cell differentiation

Differential nuclear localization of protein kinase C isoforms in neuroblastoma × glioma hybrid cells

Expression of protein kinase C isozymes in hippocampal neurones in culture

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