Abstract:
:A novel cytochemical method for the in situ, ultrastructural localization of phospholipids in biological tissues is reported. The method is based on the enzyme-gold approach (M. Bendayan: J. Histochem. Cytochem. 29, 531, 1981). Phospholipase A2 from bee venom was adsorbed on colloidal gold particles (PLA2-gold) and applied for the specific labeling of its substrate, sn3-glycerophospholipids. The binding and enzymic competence of the PLA2-gold complex were confirmed by in vitro, preembedding experiments with erythrocytes and a crude lung surfactant preparation. The substrate specificity of the probe was assessed by labeling Epon thin sections of pure phospholipids. To test the potential applications of the PLA2-gold complex, lung and pancreatic tissues were fixed with glutaraldehyde-osmium and embedded in Epon for transmission electron microscopy (TEM). They were also prepared for critical-point-drying fracture-label (CPD-FL) replicas and thin-section fracture-label (TS-FL) specimens. On TEM thin sections incubated with PLA2-gold, all cellular membranes were labeled. The labeling density over each membrane compartment, as quantitated in lung type II pneumocytes, was classified in order of magnitude as follows: a) nuclear membranes; b) outer mitochondrial membrane and rough endoplasmic reticulm (RER); and c) Golgi complex, mitochondrial cristae and plasma membranes. In lung alveoli, the phospholipid-rich surfactant material was intensely labeled. Labeling of lung thin sections from chlorphentermine-treated rats (phospholipidosis-inducing drug) further demonstrates the reliability of PLA2-gold to label phospholipids. CPD-FL replicas and TS-FL specimens further extended the TEM observations: nuclear membranes and RER were more intensely labeled than plasma membranes. In exocrine pancreatic cells, two distinct labeling patterns were found for secretory granule membranes: sparse and dense. The specificity and reliability of the labeling were confirmed through several control experiments. The studies performed thus demonstrate the great potential of the PLA2-gold technique as a new approach to the high-resolution study of phospholipid distribution and density among biological structures.
journal_name
Eur J Cell Bioljournal_title
European journal of cell biologyauthors
Coulombe PA,Kan FW,Bendayan Msubject
Has Abstractpub_date
1988-08-01 00:00:00pages
564-76issue
3eissn
0171-9335issn
1618-1298journal_volume
46pub_type
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journal_title:European journal of cell biology
pub_type: 杂志文章
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
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journal_title:European journal of cell biology
pub_type: 杂志文章
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journal_title:European journal of cell biology
pub_type: 杂志文章
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1078/0171-9335-00434
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1990-12-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1992-06-01 00:00:00
abstract::Lysosomal Ca2+ release channel TRPML1 has been suggested to regulate lysosome size by activating calmodulin (CaM). To further understand how TRPML1 and CaM regulate lysosome size, in this study, we report that inhibiting mTORC1 causes enlarged lysosomes, and the recovery of enlarged lysosomes is suppressed by inhibiti...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/j.ejcb.2019.05.001
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1016/S0171-9335(99)80053-3
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1984-05-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1983-11-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:10.1078/0171-9335-00136
更新日期:2001-01-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00
abstract::Pretreatment of Acanthamoeba castellanii (Neff strain) with sublytic concentrations of surface-active alkyltrimethylammonium salts (C12, C14, C16) enhanced ConA-mediated agglutination of the amoebae. Treatment with the surfactants alone did not affect the "spontaneous" agglutination of the amoebae. Electron microscopi...
journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1981-08-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-01-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1985-05-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1985-07-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1984-05-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
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journal_title:European journal of cell biology
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doi:
更新日期:1994-02-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1983-01-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1979-08-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1990-12-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1983-05-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
更新日期:1983-07-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
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更新日期:1998-10-01 00:00:00
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journal_title:European journal of cell biology
pub_type: 杂志文章
doi:
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