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 language | English |
|---|---|
| Pages (from-to) | 181-191 |
| Number of pages | 11 |
| Journal | Journal of Histochemistry and Cytochemistry |
| Volume | 43 |
| Issue number | 2 |
| Publication status | Published - 1995 |
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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
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 journal › Article
}
TY - JOUR
T1 - Immunocytochemical detection of phosphatidylinositol 4,5-bisphosphate localization sites within the nucleus
AU - Mazzotti, G.
AU - Zini, N.
AU - Rizzi, E.
AU - Rizzoli, R.
AU - Galanzi, A.
AU - Ognibene, A.
AU - Santi, S.
AU - Matteucci, A.
AU - Martelli, A. M.
AU - Maraldi, N. M.
PY - 1995
Y1 - 1995
N2 - 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.
AB - 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.
KW - 3T3 cells
KW - Friend cells
KW - Immunogold labeling
KW - Isolated nuclei
KW - Nuclear matrix
KW - Phosphatidylinositol 4,5-bisphosphate
KW - Rat pancreas
KW - Signal transduction
UR - http://www.scopus.com/inward/record.url?scp=0028942224&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028942224&partnerID=8YFLogxK
M3 - Article
C2 - 7822774
AN - SCOPUS:0028942224
VL - 43
SP - 181
EP - 191
JO - Journal of Histochemistry and Cytochemistry
JF - Journal of Histochemistry and Cytochemistry
SN - 0022-1554
IS - 2
ER -