Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus

G. Mazzotti, N. Zini, E. Rizzi, R. Rizzoli, A. Galanzi, A. Ognibene, S. Santi, A. Matteucci, A. M. Martelli, N. M. Maraldi

Research output: Contribution to journalArticle

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Abstract

Phosphatidylinositol 4,5-bisphosphate {PIP2) is a key element of signal transduction, being the preferential substrate of specific phospholipase that produce second messengers such as inositol trisphosphate (IP3) and diacylglycerol (DG). Because PIP2 has been cytochemically identified by monoclonal antibodies not only in the cytoplasmic membranes but also in the nuclear envelope and within the nucleus, we performed a study by immunoblotting and by confocal and electron microscopic immunocytochemistry to identify the nuclear sites of PIP2 localization and to exclude any cross- reactivity of the antibody with other nuclear molecules. The results confirm the specificity of the immunolabeling and indicate that PIP2 is localized at precise intranuclear sites both in in situ and in isolated nuclei. They also show that a significant amount of the phospholipid is retained by the cytoskeleton and by the inner nuclear matrix in in situ matrix preparations. Moreover the sensitivity of the immunocytochemical reaction is capable of detecting quantitative variations of PIP2 nuclear content induced by agonists that modulate the signal transduction system at the nuclear level.

Original languageEnglish
Pages (from-to)181-191
Number of pages11
JournalJournal of Histochemistry and Cytochemistry
Volume43
Issue number2
Publication statusPublished - 1995

Fingerprint

Phosphatidylinositols
Signal Transduction
Nuclear Matrix
Phospholipases
Diglycerides
Nuclear Envelope
Second Messenger Systems
Inositol
Cytoskeleton
Immunoblotting
Phospholipids
Immunohistochemistry
Monoclonal Antibodies
Cell Membrane
Electrons
Antibodies

Keywords

  • 3T3 cells
  • Friend cells
  • Immunogold labeling
  • Isolated nuclei
  • Nuclear matrix
  • Phosphatidylinositol 4,5-bisphosphate
  • Rat pancreas
  • Signal transduction

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Mazzotti, G., Zini, N., Rizzi, E., Rizzoli, R., Galanzi, A., Ognibene, A., ... Maraldi, N. M. (1995). Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus. Journal of Histochemistry and Cytochemistry, 43(2), 181-191.

Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus. / Mazzotti, G.; Zini, N.; Rizzi, E.; Rizzoli, R.; Galanzi, A.; Ognibene, A.; Santi, S.; Matteucci, A.; Martelli, A. M.; Maraldi, N. M.

In: Journal of Histochemistry and Cytochemistry, Vol. 43, No. 2, 1995, p. 181-191.

Research output: Contribution to journalArticle

Mazzotti, G, Zini, N, Rizzi, E, Rizzoli, R, Galanzi, A, Ognibene, A, Santi, S, Matteucci, A, Martelli, AM & Maraldi, NM 1995, 'Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus', Journal of Histochemistry and Cytochemistry, vol. 43, no. 2, pp. 181-191.
Mazzotti, G. ; Zini, N. ; Rizzi, E. ; Rizzoli, R. ; Galanzi, A. ; Ognibene, A. ; Santi, S. ; Matteucci, A. ; Martelli, A. M. ; Maraldi, N. M. / Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus. In: Journal of Histochemistry and Cytochemistry. 1995 ; Vol. 43, No. 2. pp. 181-191.
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