Structure of membrane holes in osmotic and saponin hemolysis.

The Journal of cell biology ·Vol. 56 ·No. 2 ·1973-02-00 ·Pages 519-27

Seeman P, Cheng D, Iles GH

Abstract

Serial section electron microscopy of hemolysing erythrocytes (fixed at 12 s after the onset of osmotic hemolysis) revealed long slits and holes in the membrane, extending to around 1 microm in length. Many but not all of the slits and holes (about 100-1000 A wide) were confluent with one another. Ferritin and colloidal gold (added after fixation) only permeated those cells containing membrane defects. No such large holes or slits were seen in saponin-treated erythrocytes, and the membrane was highly invaginated, giving the ghost a scalloped outline. Freeze-etch electron microscopy of saponin-treated membranes revealed 40-50 A-wide pits in the extracellular surface of the membrane. If these pits represent regions from which cholesterol was extracted, then cholesterol is uniformly distributed over the entire erythrocyte membrane.

MeSH Terms

Aldehydes Cell Membrane/analysis,drug effects Cell Membrane Permeability Cholesterol/isolation & purification Erythrocytes/cytology,drug effects Ferritins/isolation & purification Freeze Etching Gold/isolation & purification Hemolysis/drug effects Histocytochemistry Histological Techniques Humans Microscopy, Electron Osmotic Fragility Saponins/pharmacology

Chemicals

Aldehydes Saponins Gold Ferritins Cholesterol

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Seeman P

Cheng D

Iles G H

References (29)

29 references, click to expand

Simple methods for "staining with lead" at high pH in electron microscopy.
J Biophys Biochem Cytol. 1961 Dec;11:729-32 PMID: 14454024
The minimum energy of bending as a possible explanation of the biconcave shape of the human red blood cell.
J Theor Biol. 1970 Jan;26(1):61-81 PMID: 5411112
Action of saponin on biological cell membranes.
Nature. 1962 Dec 8;196:952-5 PMID: 13966357
ERYTHROCYTE METABOLISM. VI. CELL SHAPE AND THE LOCATION OF CHOLESTEROL IN THE ERYTHROCYTE MEMBRANE.
J Lab Clin Med. 1965 May;65:756-74 PMID: 14281372
[Experimental loading of erythrocyte stromata with hemoglobin; a contribution to the problem of the reversion of hemolysis].
Klin Wochenschr. 1956 Jan 15;34(3-4):101-2 PMID: 13307872
Accumulation of dextran in human red cells after haemolysis.
Nature. 1959 Aug 29;184(Suppl 10):723-4 PMID: 14421492
The erythrocyte ghost is a perfect osmometer.
J Gen Physiol. 1970 Feb;55(2):208-19 PMID: 5413078
Ultrastructure of red cell membranes.
Semin Hematol. 1970 Jul;7(3):259-74 PMID: 4913045
Demonstration of the outer surface of freeze-etched red blood cell membranes.
J Cell Biol. 1970 Jun;45(3):649-53 PMID: 4918217
The entry of ferritin into hemoglobin-free human erythrocyte ghosts prepared under different conditions.
J Ultrastruct Res. 1970 Sep;32(5):405-16 PMID: 5478685
The arrangement of subunits in the shell of ferritin. Electron microscopic observations of molecules negatively stained with uranyl acetate.
J Ultrastruct Res. 1970 Dec;33(5):442-50 PMID: 4099128
Electron microscopic studies of lipid protein films.
Nature. 1971 May 28;231(5300):267-8 PMID: 4930694
[The erythrocyte membrane. Fine structure of the freeze etched membrane after treatment with hypotonic solutions and saponin].
Klin Wochenschr. 1970 Aug 1;48(15):939-43 PMID: 4933166
The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies.
J Cell Biol. 1971 Oct;51(1):265-72 PMID: 5000071
Synaptic vesicles in freeze-etched electric tissue of Torpedo.
Philos Trans R Soc Lond B Biol Sci. 1971 Jun 17;261(839):383-5 PMID: 4399629
Translational mobility of the membrane intercalated particles of human erythrocyte ghosts. pH-dependent, reversible aggregation.
J Cell Biol. 1972 Jun;53(3):777-87 PMID: 4554989
A study of fixation for electron microscopy.
J Exp Med. 1952 Mar;95(3):285-98 PMID: 14927794
Physiological characteristics of human red blood cell ghosts.
J Gen Physiol. 1958 Sep 20;42(1):9-28 PMID: 13575771
Osmotic hemolysis by a gradual decrease in the ionic strength of the surrounding medium.
J Cell Comp Physiol. 1961 Apr;57:111-7 PMID: 13719676
Electron microscopic observations on Rous sarcoma virus and cell membranes.
Nature. 1962 Jun 23;194:1116-9 PMID: 13887581
Cell junctions in amphibian skin.
J Cell Biol. 1965 Jul;26(1):263-91 PMID: 5859021
The iron oxide core of the ferritin molecule.
J Mol Biol. 1965 Dec;14(2):598-602 PMID: 5880869
A simple freeze-fracture replication method for electron microscopy.
J Cell Biol. 1966 Jun;29(3):435-47 PMID: 5962938
The loss of certain cellular components from human erythrocytes during hypotonic hemolysis in the presence of dextran.
Acta Physiol Scand. 1966 May;67(1):43-9 PMID: 5963301
Entry of ferritin into human red cells during hypotonic haemolysis.
Nature. 1967 Jul 22;215(5099):424-5 PMID: 6058309
Ultrastructure of the red blood cell.
Fed Proc. 1967 Nov-Dec;26(6):1785-801 PMID: 4169459
The proton relaxation of benzyl alcohol in erythrocyte membranes.
Mol Pharmacol. 1968 Jan;4(1):87-95 PMID: 5640782
Visualization of substructure in ferritin molecules: an artifact.
J Microsc. 1969;89(2):251-61 PMID: 5820004
Structure of red-cell ghosts and the effect of saponin treatment.
Nature. 1963 Feb 23;197:822 PMID: 13955901

Article Info

Journal

The Journal of cell biology

Abbr.

J Cell Biol

ISSN

0021-9525

DOI

10.1083/jcb.56.2.519

Published

1973-02-00

Pages

519-27

Language

English

Region

United States

NLM ID

0375356

PMCID

PMC2108893

Subset

External Links

PubMed source DOI: 10.1083/jcb.56.2.519 PMC Full Text View journal details

Analysis Services

Structure of membrane holes in osmotic and saponin hemolysis.

Contact

WeChat Official Account