• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Oct 1978; 75(10): 4977–4981.
PMCID: PMC336245

Partial purification and characterization of the intercellular bridge from cultured mouse cells

Abstract

At the completion of mitosis, the two daughter cells are connected by a channel of membrane-bound cytoplasm, the intercellular bridge. This structure contains parallel arrays of spindle microtubules which are associated, at the bridge midline, with a metallophilic band called the midbody. In an effort to characterize midbody components, intercellular bridges were partially purified. The purification consisted of brief sonication of telophase populations of mouse L929 cells in order to shear intercellular bridges from daughter cells, digestion of chromatin by an excess of micrococcal nuclease, and differential centrifugation to enrich for intercellular bridges. Electron microscopy of these preparations substantiated the identity of the bulk of material as intercellular bridges. After solubilization with sodium dodecyl sulfate, the protein components of these preparations were iodinated with Na125I and separated by two-dimensional gel electrophoresis. From these analyses, the major polypeptide components of intercellular bridges appear to be tubulin, varying amounts of plasma membrane proteins, and a polypeptide with an apparent molecular weight of 42,000. Time-course studies reveal that this polypeptide is first detectable in a pelletable form approximately 30 min after cells are released from metaphase block, reaches maximal spot intensity in late telophase, and is no longer detectable in G1 populations. We interpret these data to suggest that the 42,000-dalton polypeptide is a component of the midbody.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • KRISHAN A, BUCK RC. STRUCTURE OF THE MITOTIC SPINDLE IN L STRAIN FIBROBLASTS. J Cell Biol. 1965 Mar;24:433–444. [PMC free article] [PubMed]
  • BUCK RC, TISDALE JM. The fine structure of the mid-body of the rat erythroblast. J Cell Biol. 1962 Apr;13:109–115. [PMC free article] [PubMed]
  • Mullins JM, Biesele JJ. Cytokinetic activities in a human cell line: the midbody and intercellular bridge. Tissue Cell. 1973;5(1):47–61. [PubMed]
  • ROBBINS E, GONATAS NK. THE ULTRASTRUCTURE OF A MAMMALIAN CELL DURING THE MITOTIC CYCLE. J Cell Biol. 1964 Jun;21:429–463. [PMC free article] [PubMed]
  • Erlandson RA, de Harven E. The ultrastructure of synchronized HeLa cells. J Cell Sci. 1971 Mar;8(2):353–397. [PubMed]
  • Mullins JM, Biesele JJ. Terminal phase of cytokinesis in D-98s cells. J Cell Biol. 1977 Jun;73(3):672–684. [PMC free article] [PubMed]
  • Paweletz N. Zur Funktion des "Flemming-Körpers" bei der Teilung tierischer Zellen. Naturwissenschaften. 1967 Oct;54(20):533–535. [PubMed]
  • Allenspach AL, Roth LE. Structural variations during mitosis in the chick embryo. J Cell Biol. 1967 Apr;33(1):179–196. [PMC free article] [PubMed]
  • Jones OP. Elimination of midbodies from mitotic erythroblasts and their contribution to fetal blood plasma. J Natl Cancer Inst. 1969 May;42(5):753–759. [PubMed]
  • Miller OL, Jr, Beatty BR. Visualization of nucleolar genes. Science. 1969 May 23;164(3882):955–957. [PubMed]
  • Miller OL, Jr, Hamkalo BA, Thomas CA., Jr Visualization of bacterial genes in action. Science. 1970 Jul 24;169(3943):392–395. [PubMed]
  • Miller OL, Jr, Bakken AH. Morphological studies of transcription. Acta Endocrinol Suppl (Copenh) 1972;168:155–177. [PubMed]
  • GREENWOOD FC, HUNTER WM, GLOVER JS. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. [PMC free article] [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Cande WZ, Lazarides E, McIntosh JR. A comparison of the distribution of actin and tubulin in the mammalian mitotic spindle as seen by indirect immunofluorescence. J Cell Biol. 1977 Mar;72(3):552–567. [PMC free article] [PubMed]
  • Rattner JB, Krystal G, Hamkalo BA. Selective digestion of mouse metaphase chromosomes. Chromosoma. 1978 Apr 25;66(3):259–268. [PubMed]
  • Rattner JB. Nuclear shaping in marsupial spermatids. J Ultrastruct Res. 1972 Sep;40(5):498–512. [PubMed]
  • Hepler PK, Jackson WT. Microtubules and early stages of cell-plate formation in the endosperm of Haemanthus katherinae Baker. J Cell Biol. 1968 Aug;38(2):437–446. [PMC free article] [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...