Abstract
The freeze-fracture, freeze-etch technique can be employed to reveal new details of the process of fusion of two unit membranes For this study, mucocyst discharge in Tetrahymena pyriformis provides a model system with certain general implications The undischarged mature mucocyst is a saclike, membrane-bound, secretory vesicle containing crystalline material The organelle tip finds its way toward a special site, a rosette of 150 Å diameter particles within the plasma membrane. To match this site, the mucocyst membrane forms an annulus of 110 Å diameter particles, above whose inner edge the rosette particles sit. Discharge of some mucocysts is triggered by fixation. As discharge proceeds, the organelle becomes spherical and its content changes from crystalline to amorphous. The cytoplasm between the two matching membrane sites is squeezed away and the membranes fuse Steps in membrane reorganization can be reconstructed from changes in rosette appearance in the fracture faces. First, a depression in the rosette—the fusion pocket—forms. The rosette particles spread at the lip as the pocket deepens and enlarges from 60 to 200 nm. The annulus particles then become visible at the lip, indicating completed fusion of the A fracture faces of mucocyst and plasma membranes The remaining B faces of the two membranes have opposite polarities When the content of the mucocyst is released, the edges of these faces join so that the unit membrane runs uninterruptedly around the lip and into the pocket.
Full Text
The Full Text of this article is available as a PDF (3.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Akert K., Moor H., Pfenninger K., Sandri C. Contributions of new impregnation methods and freeze etching to the problems of synaptic fine structure. Prog Brain Res. 1969;31:223–240. doi: 10.1016/S0079-6123(08)63241-0. [DOI] [PubMed] [Google Scholar]
- Allen R. D. Fine structure, reconstruction and possible functions of components of the cortex of Tetrahymena pyriformis. J Protozool. 1967 Nov;14(4):553–565. doi: 10.1111/j.1550-7408.1967.tb02042.x. [DOI] [PubMed] [Google Scholar]
- Bodian D. An electron microscopic characterization of classes of synaptic vesicles by means of controlled aldehyde fixation. J Cell Biol. 1970 Jan;44(1):115–124. doi: 10.1083/jcb.44.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Branton D. Fracture faces of frozen membranes. Proc Natl Acad Sci U S A. 1966 May;55(5):1048–1056. doi: 10.1073/pnas.55.5.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chalcroft J. P., Bullivant S. An interpretation of liver cell membrane and junction structure based on observation of freeze-fracture replicas of both sides of the fracture. J Cell Biol. 1970 Oct;47(1):49–60. doi: 10.1083/jcb.47.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deamer D. W., Branton D. Fracture planes in an ice-bilayer model membrane system. Science. 1967 Nov 3;158(3801):655–657. doi: 10.1126/science.158.3801.655. [DOI] [PubMed] [Google Scholar]
- Deamer D. W., Leonard R., Tardieu A., Branton D. Lamellar and hexagonal lipid phases visualized by freeze-etching. Biochim Biophys Acta. 1970;219(1):47–60. doi: 10.1016/0005-2736(70)90060-x. [DOI] [PubMed] [Google Scholar]
- Frye L. D., Edidin M. The rapid intermixing of cell surface antigens after formation of mouse-human heterokaryons. J Cell Sci. 1970 Sep;7(2):319–335. doi: 10.1242/jcs.7.2.319. [DOI] [PubMed] [Google Scholar]
- Gilula N. B., Branton D., Satir P. The septate junction: a structural basis for intercellular coupling. Proc Natl Acad Sci U S A. 1970 Sep;67(1):213–220. doi: 10.1073/pnas.67.1.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilula N. B., Satir P. The ciliary necklace. A ciliary membrane specialization. J Cell Biol. 1972 May;53(2):494–509. doi: 10.1083/jcb.53.2.494. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leak L. V. Frozen-fractured images of blood capillaries in heart tissue. J Ultrastruct Res. 1971 Apr;35(1):127–146. doi: 10.1016/s0022-5320(71)80147-8. [DOI] [PubMed] [Google Scholar]
- Nanney D. L. The pattern of replication of cortical units in Tetrahymena. Dev Biol. 1971 Oct;26(2):296–305. doi: 10.1016/0012-1606(71)90128-x. [DOI] [PubMed] [Google Scholar]
- Nickel E., Grieshaber E. Elektronemikroskopische Darstellung der Muskelkapillaren im Gefrierätzbild. Z Zellforsch Mikrosk Anat. 1969;95(3):445–461. [PubMed] [Google Scholar]
- Palade G. E., Bruns R. R. Structural modulations of plasmalemmal vesicles. J Cell Biol. 1968 Jun;37(3):633–649. doi: 10.1083/jcb.37.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pinto da Silva P., Branton D. Membrane splitting in freeze-ethching. Covalently bound ferritin as a membrane marker. J Cell Biol. 1970 Jun;45(3):598–605. doi: 10.1083/jcb.45.3.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pinto da Silva P. Translational mobility of the membrane intercalated particles of human erythrocyte ghosts. pH-dependent, reversible aggregation. J Cell Biol. 1972 Jun;53(3):777–787. doi: 10.1083/jcb.53.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROBERTSON J. D. The ultrastructure of cell membranes and their derivatives. Biochem Soc Symp. 1959;16:3–43. [PubMed] [Google Scholar]
- Satir B., Dirksen E. R. Nucleolar aging in Tetrahymena during the cultural growth cycle. J Cell Biol. 1971 Jan;48(1):143–154. doi: 10.1083/jcb.48.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tokuyasu K., Scherbaum O. H. Ultrastructure of mucocysts and pellicle of Tetrahymena pyriformis. J Cell Biol. 1965 Oct;27(1):67–81. doi: 10.1083/jcb.27.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WILLIAMS N. E., SCHERBAUM O. H. Morphogenetic events in normal and synchronously dividing Tetrahymena. J Embryol Exp Morphol. 1959 Jun;7:241–256. [PubMed] [Google Scholar]
