Skip to content

Research at St Andrews

Phospholipase C-η2 interacts with nuclear and cytoplasmic LIMK-1 during retinoic acid-stimulated neurite growth

Research output: Contribution to journalArticle

Author(s)

Mohammed Arastoo, Christian Hacker, Petra Popovics, John Milton Lucocq, Alan J. Stewart

School/Research organisations

Abstract

Neurite growth is central to the formation and differentiation of functional neurons and recently, an essential role for phospholipase C-η2 (PLCη2) in neuritogenesis was revealed. Here we investigate the function of PLCη2 in neuritogenesis using Neuro2A cells, which upon stimulation with retinoic acid differentiate and form neurites. We first investigated the role of the PLCη2 calcium-binding EF-hand domain, a domain that is known to be required for calcium-dependent PLCη2 activation. To do this we quantified neurite outgrowth in Neuro2A cells, stably overexpressing wild-type PLCη2 and D256A (EF-hand) and H460Q (active site) PLCη2 mutants. Retinoic acid-induced neuritogenesis was highly dependent on PLCη2 activity, with the H460Q mutant exhibiting a strong dominant-negative effect. Expression of the D256A mutant had little effect on neurite growth relative to the control, suggesting that calcium-directed activation of PLCη2 is not essential to this process. We next investigated which cellular compartments contain endogenous PLCη2 by comparing immuno-electron microscopy signals over control and knockdown cell lines. When signals were analyzed to reveal specific labelling for PLCη2, it was found to be localized predominantly over the nucleus and cytosol. Furthermore in these compartments (and also in growing neurites), a proximity ligand assay revealed that PLCη2 specifically interacts with LIMK-1 in Neuro2A cells. Taken together, these data emphasize the importance of PLCη2 and its articulation with LIMK-1 in regulating neuritogenesis.
Close

Details

Original languageEnglish
Pages (from-to)163-173
JournalHistochemistry and Cell Biology
Volume145
Issue number2
Early online date15 Dec 2015
DOIs
Publication statusPublished - Feb 2016

    Research areas

  • Calcium signaling, Cell differentiation, Electron microscopy, Neuritogenesis, Protein-protein interaction

Discover related content
Find related publications, people, projects and more using interactive charts.

View graph of relations

Related by author

  1. The X files: ''The mystery of X chromosome instability in Alzheimer's disease''

    Bajic, V. P., Essak, M., Zivkovic, L., Stewart, A. J., Zafirovic, S., Bajic, V., Gojobori, T., Isenovic, E. & Spremo-Potparevic, B., 13 Dec 2019, (Accepted/In press) In : Frontiers in Genetics.

    Research output: Contribution to journalReview article

  2. Coagulatory defects in type-1 and type-2 diabetes

    Sobczak, A. I. S. & Stewart, A. J., 12 Dec 2019, (Accepted/In press) In : International Journal of Molecular Sciences.

    Research output: Contribution to journalReview article

  3. Reduced plasma magnesium levels in type-1 diabetes associate with prothrombotic changes in fibrin clotting and fibrinolysis

    Sobczak, A. I. S., Phoenix, F. A., Pitt, S. J., Ajjan, R. A. & Stewart, A. J., 11 Nov 2019, (Accepted/In press) In : Thrombosis and Haemostasis.

    Research output: Contribution to journalArticle

  4. A metalloproteomic analysis of interactions between plasma proteins and zinc: elevated fatty acid levels affect zinc distribution

    Coverdale, J. P. C., Barnett, J. P., Adamu, A. H., Griffiths, E. J., Stewart, A. J. & Blindauer, C. A., 1 Nov 2019, In : Metallomics. 11, 11, p. 1805-1819 15 p.

    Research output: Contribution to journalArticle

  5. Plasma non-esterified fatty acids contribute to increased coagulability in type-2 diabetes through altered plasma zinc speciation

    Sobczak, A. I. S., Katundu, K. G. H., Phoenix, F. A., Khazaipoul, S., Yu, R., Lampiao, F., Stefanowicz, F., Blindauer, C. A., Pitt, S. J., Smith, T. K., Ajjan, R. A. & Stewart, A. J., 28 Aug 2019, In : biorxiv. 38 p.

    Research output: Contribution to journalArticle

Related by journal

  1. Quantifying Golgi structure using EM: combining volume-SEM and stereology for higher throughput

    Ferguson, S., Steyer, A. M., Mayhew, T. M., Schwab, Y. & Lucocq, J. M., Jun 2017, In : Histochemistry and Cell Biology. 147, 6, p. 653-669 17 p.

    Research output: Contribution to journalReview article

  2. Antibodies for immunolabeling by light and electron microscopy: not for the faint hearted

    Griffiths, G. & Lucocq, J. M., Oct 2014, In : Histochemistry and Cell Biology. 142, 4, p. 347-360 14 p.

    Research output: Contribution to journalReview article

  3. Developments in cell biology for quantitative immunoelectron microscopy based on thin sections: a review

    Mayhew, T. M. & Lucocq, J. M., Aug 2008, In : Histochemistry and Cell Biology. 130, 2, p. 299-313 15 p.

    Research output: Contribution to journalReview article

ID: 233941196

Top