PMID- 17011819
OWN - NLM
STAT- MEDLINE
DCOM- 20061122
LR  - 20161126
IS  - 0006-3002 (Print)
IS  - 0006-3002 (Linking)
VI  - 1761
IP  - 10
DP  - 2006 Oct
TI  - Molecular probes for sensing the cholesterol composition of subcellular organelle 
      membranes.
PG  - 1169-81
AB  - Neuroendocrine cells contain two types of secretagogue-regulated acidic 
      compartments: secretory granules (SGs) and synaptic-like microvesicles (SLMVs), 
      which can be identified by acidotropic probes such as acridine orange (AO) and 
      DAMP. We investigated the accumulation of these probes in SGs and SLMVs as a 
      function of glucose levels in the culture media using a pancreatic beta-cell line 
      MIN6. AO was accumulated in the low-glucose condition, but not in the 
      high-glucose condition. The AO accumulation correlated well with the SLMV 
      dynamics by glucose and DAMP was localized in the SGs. Because SG membranes are 
      reportedly high in cholesterol, we prepared liposomes with increasing cholesterol 
      levels. AO is well incorporated into liposomes having a 20 to 40 mol% cholesterol 
      composition, whereas DAMP was so in those having over 40 mol% cholesterol levels. 
      Indeed, when cholesterol was depleted from MIN6 SG membranes, DAMP incorporation 
      decreased, instead AO was incorporated. In PC12 cells, AO incorporation into SGs 
      was significant but DAMP incorporation was limited. Consistently, the cholesterol 
      composition was found 37 to 39 mol% in the SG membrane of PC12 cells. We suggest 
      that cholesterol-sensing probes, AO and DAMP, are useful tools for investigating 
      cholesterol compositions in acidic organelle membranes.
FAU - Wang, Rong
AU  - Wang R
AD  - Department of Molecular Medicine, Institute for Molecular and Cellular 
      Regulation, Gunma University, Showa-machi, Maebashi 371-8512, Japan.
FAU - Hosaka, Masahiro
AU  - Hosaka M
FAU - Han, Lu
AU  - Han L
FAU - Yokota-Hashimoto, Hiromi
AU  - Yokota-Hashimoto H
FAU - Suda, Masayuki
AU  - Suda M
FAU - Mitsushima, Dai
AU  - Mitsushima D
FAU - Torii, Seiji
AU  - Torii S
FAU - Takeuchi, Toshiyuki
AU  - Takeuchi T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060830
PL  - Netherlands
TA  - Biochim Biophys Acta
JT  - Biochimica et biophysica acta
JID - 0217513
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Liposomes)
RN  - 0 (Molecular Probes)
RN  - 92585-03-0 (3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine)
RN  - 97C5T2UQ7J (Cholesterol)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/metabolism
MH  - Animals
MH  - Cell Line
MH  - Cholesterol/*analysis
MH  - Dinitrobenzenes/metabolism
MH  - Fluorescent Dyes/metabolism
MH  - Intracellular Membranes/*chemistry
MH  - Islets of Langerhans
MH  - Liposomes/metabolism
MH  - Mice
MH  - *Molecular Probes
MH  - Organelles/ultrastructure
MH  - PC12 Cells
MH  - Rats
MH  - Secretory Vesicles/chemistry
EDAT- 2006/10/03 09:00
MHDA- 2006/12/09 09:00
CRDT- 2006/10/03 09:00
PHST- 2006/02/21 00:00 [received]
PHST- 2006/06/14 00:00 [revised]
PHST- 2006/06/14 00:00 [accepted]
PHST- 2006/10/03 09:00 [pubmed]
PHST- 2006/12/09 09:00 [medline]
PHST- 2006/10/03 09:00 [entrez]
AID - S1388-1981(06)00234-4 [pii]
AID - 10.1016/j.bbalip.2006.06.016 [doi]
PST - ppublish
SO  - Biochim Biophys Acta. 2006 Oct;1761(10):1169-81. doi: 
      10.1016/j.bbalip.2006.06.016. Epub 2006 Aug 30.