Abstract
Nuclear Ca2+ regulates key cellular processes, including gene expression, apoptosis, assembly of the nuclear envelope, and nucleocytoplasmic transport. Quantifi cation of subcellularly resolved Ca2+ signals is, therefore, essential for understanding physiological and pathological processes in various cell types. However, the properties of commonly used Ca2+ -fl uorescent indicators in intracellular compartments may differ, thus affecting the translation of Ca2+ -dependent fl uorescence changes into quantitative changes of Ca2+ concentration. Here, we describe technical approaches for reliable subcellular quantifi cation of [Ca2+] in the cytoplasm vs. the nucleus and the nuclear envelope by in situ calibration of fl uorescein-derived fl uorescent indicators Fluo-4 and Fluo-5N.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 135-147 |
| Number of pages | 13 |
| Journal | Methods in Molecular Biology |
| Volume | 1234 |
| DOIs | |
| State | Published - 2015 |
Fingerprint
Keywords
- Ca concentration
- Cytoplasm
- Fluo-4
- Fluo-5N
- In situ calibration
- Nuclear envelope
- Nucleus
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Measuring intranuclear and nuclear envelope [ca2+] vs. cytosolic [ca2+]. / Ljubojević, Senka; Bers, Donald M.
In: Methods in Molecular Biology, Vol. 1234, 2015, p. 135-147.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Measuring intranuclear and nuclear envelope [ca2+] vs. cytosolic [ca2+]
AU - Ljubojević, Senka
AU - Bers, Donald M.
PY - 2015
Y1 - 2015
N2 - Nuclear Ca2+ regulates key cellular processes, including gene expression, apoptosis, assembly of the nuclear envelope, and nucleocytoplasmic transport. Quantifi cation of subcellularly resolved Ca2+ signals is, therefore, essential for understanding physiological and pathological processes in various cell types. However, the properties of commonly used Ca2+ -fl uorescent indicators in intracellular compartments may differ, thus affecting the translation of Ca2+ -dependent fl uorescence changes into quantitative changes of Ca2+ concentration. Here, we describe technical approaches for reliable subcellular quantifi cation of [Ca2+] in the cytoplasm vs. the nucleus and the nuclear envelope by in situ calibration of fl uorescein-derived fl uorescent indicators Fluo-4 and Fluo-5N.
AB - Nuclear Ca2+ regulates key cellular processes, including gene expression, apoptosis, assembly of the nuclear envelope, and nucleocytoplasmic transport. Quantifi cation of subcellularly resolved Ca2+ signals is, therefore, essential for understanding physiological and pathological processes in various cell types. However, the properties of commonly used Ca2+ -fl uorescent indicators in intracellular compartments may differ, thus affecting the translation of Ca2+ -dependent fl uorescence changes into quantitative changes of Ca2+ concentration. Here, we describe technical approaches for reliable subcellular quantifi cation of [Ca2+] in the cytoplasm vs. the nucleus and the nuclear envelope by in situ calibration of fl uorescein-derived fl uorescent indicators Fluo-4 and Fluo-5N.
KW - Ca concentration
KW - Cytoplasm
KW - Fluo-4
KW - Fluo-5N
KW - In situ calibration
KW - Nuclear envelope
KW - Nucleus
UR - http://www.scopus.com/inward/record.url?scp=84921745693&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84921745693&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-1755-6_12
DO - 10.1007/978-1-4939-1755-6_12
M3 - Article
C2 - 25304354
AN - SCOPUS:84921745693
VL - 1234
SP - 135
EP - 147
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -