Programmed cell death (PCD) of Dictyostelium discoideum cells was triggered precisely and studied quantitatively in an in vitro system involving differentiation without morphogenesis. In temporal succession after the triggering of differentiation, PCD included first an irreversible step leading to the inability to regrow at 8 hours. At 12 hours, massive vacuolisation was best evidenced by confocal microscopy, and prominent cytoplasmic condensation and focal chromatin condensation could be observed by electron microscopy. Membrane permeabilization occurred only very late (at 40–60 hours) as judged by propidium iodide staining. No early DNA fragmentation could be detected by standard or pulsed field gel electrophoresis. These traits exhibit some similarity to those of previously described non-apoptotic and apoptotic PCD, suggesting the hypothesis of a single core molecular mechanism of PCD emerging in evolution before the postulated multiple emergences of multicellularity. A single core mechanism would underly phenotypic variations of PCD resulting in various cells from differences in enzymatic equipment and mechanical constraints. A prediction is that some of the molecules involved in the core PCD mechanism of even phylogenetically very distant organisms, e.g. Dictyostelium and vertebrates, should be related.

REFERENCES

Abrams
J. M.
,
White
K.
,
Fessler
L. I.
,
Steller
H.
(
1993
).
Programmed cell death during Drosophila embryogenesis.
Development
117
,
29
43
Berks
M.
,
Kay
R. R.
(
1988
).
Cyclic AMP is an inhibitor of stalk cell differentiation in Dictyostelium discoideum.
Dev. Biol
126
,
108
114
Blumberg
D. D.
,
Agarwal
A. K.
,
Sloger
M. S.
,
Yoder
B. K.
(
1991
).
Gene expression and chromatin structure in the cellular slime mold, Dictyostelium discoideum.
Dev. Gen
12
,
65
77
Brown
D. G.
,
Sun
X.-M.
,
Cohen
G. M.
) (
1993
).
Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation.
J. Biol. Chem
268
,
3037
3039
Christen
R.
,
Ratto
A.
,
Baroin
A.
,
Perasso
R.
,
Grell
K. G.
,
Adoutte
A.
(
1991
).
An analysis of the origin of metazoans, using comparisons of partial sequences of the 28S RNA, reveals an early emergence of triploblasts.
EMBO J
10
,
499
503
Clarke
P. G. H.
(
1990
).
Developmental cell death: morphological diversity and multiple mechanisms.
Anat. Embryol
181
,
195
213
Cockburn
A. F.
,
Taylor
W. C.
,
Firtel
R. A.
(
1978
).
Dictyostelium rDNA consists of non-chromosomal palindromic dimers containing 5S and 36S coding regions.
Chromosoma
70
,
19
29
Cohen
J. J.
,
Duke
R. C.
(
1984
).
Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death.
J. Immunol
132
,
38
42
Cohen
J. J.
(
1991
).
Programmed cell death in the immune system.
Advan. Immunol
50
,
55
83
Cox
E. C.
,
Vocke
C. D.
,
Walter
S.
,
Gregg
K. Y.
,
Bain
E. S.
(
1990
).
Electrophoretic karyotype for Dictyostelium discoideum.
Proc. Nat. Acad. Sci. USA
87
,
8247
8251
de Chastellier
C.
,
Ryter
A.
(
1977
).
Changes of the cell surface and of the digestive apparatus of Dictyostelium discoideum during the starvation period triggering aggregation.
J. Cell Biol
75
,
218
236
Duvall
E.
,
Wyllie
A. H.
(
1986
).
Death and the cell. Immunol.
Today
7
,
115
119
Eleftheriou
E. P.
(
1986
).
Ultrastructural studies on protophloem sieve elements in Triticum aestivum L. Nuclear degeneration.
J. Ultrastruct. Mol. Struct. Res
95
,
47
60
Ellis
H. M.
,
Horvitz
H. R.
(
1986
).
Genetic control of programmed cell death in the nematode C. elegans.
Cell
44
,
817
829
Ellis
R. E.
(
1992
).
Negative regulators of programed cell death.
Curr. Opin. Gen. Dev
2
,
635
641
Feinberg
A. P.
,
Vogelstein
B.
(
1984
).
A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity.
Analyt. Biochem
137
,
266
267
Field
K. G.
,
Olsen
G. J.
,
Lane
D. J.
,
Giovannoni
S. J.
,
Ghiselin
M. T.
,
Raff
E. C.
,
Pace
N. R.
,
Raff
R. A.
(
1988
).
Molecular phylogeny of the animal kingdom.
Science
239
,
748
753
Garcia-Martinez
V.
,
Macias
D.
,
Gañan
Y.
,
Garcia-Lobo
J. M.
,
Francia
M. V.
,
Fernandez-Teran
M. A.
,
Hurle
J. M.
(
1993
).
Internucleosomal DNA fragmentation and programmed cell death (apoptosis) in the interdigital tissue of the embryonic chick leg bud.
J. Cell Sci
106
,
201
208
George
R. P.
,
Hohl
H. R.
,
Raper
K. B.
(
1972
).
Ultrastructural development of stalk-producing cells in Dictyostelium discoideum, a cellular slime mould.
J. Gen. Microbiol
70
,
477
489
Golstein
P.
,
Ojcius
D. M.
,
Young
J. D.-E.
(
1991
).
Cell death mechanisms and the immune system.
Immunol. Rev.
121
,
29
65
Hedgecock
E. M.
,
Sulston
J. E.
,
Thomson
J. N.
(
1983
).
Mutations affecting programmed cell death in the nematode Caenorhabitis elegans.
Science
220
,
1277
1279
Humbert
M.
,
Raposo
G.
,
Cosson
P.
,
Reggio
H.
,
Davoust
J.
,
Salamero
J.
(
1993
).
The invariant chain induces compact forms of Class II molecules localized in late endosomal compartments.
Eur. J. Immunol
23
,
3158
3166
Johnson
E. M.
Jr.
,
Deckwerth
T. L.
(
1993
).
Molecular mechanisms of developmental neuronal death.
Annu. Rev. Neurosci
16
,
31
46
Kaiser
D.
(
1986
).
Control of multicellular development: Dictyostelium and Myxococcus.
Annu. Rev. Genet
20
,
539
566
Kay
R. R.
(
1987
).
Cell differentiation in monolayers and the investigation of slime mold morphogens.
Meth. Cell Biol
28
,
433
448
Kerr
J. F. R.
,
Wyllie
A. H.
,
Currie
A. R.
(
1972
).
Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics.
Br. J. Cancer
26
,
239
257
Kopachik
W.
,
Oohata
A.
,
Dhokia
B.
,
Brookman
J. J.
,
Kay
R. R.
(
1983
).
Dictyostelium mutants lacking DIF, a putative morphogen.
Cell
33
,
397
403
Kopachik
W. J.
,
Dhokia
B.
,
Kay
R. R.
(
1985
).
Selective induction of stalk-cell-specific proteins in Dictyostelium.
Differentiation
28
,
209
216
Kuspa
A.
,
Loomis
W. F.
(
1992
).
Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.
Proc. Nat. Acad. Sci. USA
89
,
8803
8807
Loomis
W. F.
(
1987
).
Genetic tools for Dictyostelium discoideum.
Meth. Cell Biol
28
,
31
65
Maeda
Y.
,
Takeuchi
I.
(
1969
).
Cell differentiation and fine structures in the development of the cellular slime molds.
Dev. Growth Differ
11
,
232
245
Michaelis
C.
,
Weeks
G.
(
1992
).
Isolation and characterization of a cdc2 cDNA from Dictyostelium discoideum.
Biochim. Biophys. Acta
1132
,
35
42
Miura
M.
,
Zhu
H.
,
Rotello
R.
,
Hartwieg
E. A.
,
Yuan
J.
(
1993
).
Induction of apoptosis in fibroblasts by IL-1-converting enzyme, a mammalian homolog of the C. elegans cell death gene ced-3.
Cell
75
,
653
660
Morris
H. R.
,
Taylor
G. W.
,
Masento
M. S.
,
Jermyn
K. A.
,
Kay
R. R.
(
1987
).
Chemical structure of the morphogen differentiation inducing factor from Dictyostelium discoideum.
Nature
328
,
811
814
Oberhammer
F.
,
Wilson
J. W.
,
Dive
C.
,
Morris
I. D.
,
Hickman
J. A.
,
Wakeling
A. E.
,
Walker
P. R.
,
Sikorska
M.
(
1993
).
Apoptotic death in epithelial cells: Cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation.
EMBO J
12
,
3679
3684
Oppenheim
R. W.
(
1991
).
Cell death during development of the nervous system.
Annu. Rev. Neurosci
14
,
453
501
Owens
G. P.
,
Hahn
W. E.
,
Cohen
J. J.
(
1991
).
Identification of mRNAs associated with programmed cell death in immature thymocytes.
Mol. Cell. Biol
11
,
4177
4188
Quiviger
B.
,
Benichou
J.-C.
,
Ryter
A.
) (
1980
).
Comparative cytochemical localization of alkaline and acid phosphatases during starvation and differentiation of Dictyostelium discoideum.
Biol. Cellulaire
37
,
241
250
Robertson
A. M. G.
,
Thomson
J. N.
(
1982
).
Morphology of programmed cell death in the ventral nerve cord of Caenorhabditis elegans larvae.
J. Embryol. Exp. Morph
67
,
89
100
Saunders
J. W.
Jr.
(
1966
).
Death in embryonic systems.
Science
154
,
604
612
Schaap
P.
,
van der Molen
L.
,
Konijn
T. M.
(
1981
).
The vacuolar apparatus of the simple cellular slime mold Dictyostelium minutum. Biol.
Cell
41
,
133
142
Schwartz
L. M.
,
Osborne
B. A.
(
1993
).
Programmed cell death, apoptosis and killer genes.
Immunol. Today
14
,
582
590
Schwartz
L. M.
,
Smith
S. W.
,
Jones
M. E. E.
,
Osborne
B. A.
(
1993
).
Do all programmed cell deaths occur via apoptosis?.
Proc. Nat. Acad. Sci. USA
90
,
980
984
Shi
L.
,
Nishioka
W. K.
,
Th'ng
J.
,
Bradbury
E. M.
,
Litchfield
D. W.
,
Greenberg
A. H.
(
1994
).
Premature p34cdc2activation required for apoptosis.
Science
263
,
1143
1145
Shimonkevitz
R.
,
Kappler
J.
,
Marrack
P.
,
Grey
H.
(
1983
).
Antigen recognition by H-2-restricted T cells. I. Cell-free antigen processing.
J. Exp. Med
158
,
303
316
Sobolewski
A.
,
Neave
N.
,
Weeks
G.
(
1983
).
The induction of stalk cell differentiation in submerged monolayers of Dictyostelium discoideum. Characterization of the temporal sequence for the molecular requirements.
Differentiation
25
,
93
100
Takeshige
K.
,
Baba
M.
,
Tsuboi
S.
,
Noda
T.
,
Ohsumi
Y.
(
1992
).
Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction.
J. Cell Biol
119
,
301
311
Tomei
L. D.
,
Shapiro
J. P.
,
Cope
F. O.
(
1993
).
Apoptosis in C3H/10T® mouse embryonic cells: Evidence for internucleosomal DNA modification in the absence of double-strand cleavage.
Proc. Nat. Acad. Sci. USA
90
,
853
857
Town
C. D.
,
Gross
J. D.
,
Kay
R. R.
(
1976
).
Cell differentiation without morphogenesis in Dictyostelium discoideum.
Nature
262
,
717
719
Town
C.
,
Stanford
E.
(
1979
).
An oligosaccharide-containing factor that induces cell differentiation in Dictyostelium discoideum.
Proc. Nat. Acad. Sci. USA
76
,
308
312
Vaux
D. L.
,
Weissman
I. L.
,
Kim
S. K.
(
1992
).
Prevention of programmed cell death in Caenorhabditis elegans by human bcl-2.
Science
258
,
1955
1957
Vaux
D. L.
(
1993
).
Toward an understanding of the molecular mechanisms of physiological cell death.
Proc. Nat. Acad. Sci. USA
90
,
786
789
Walker
P. R.
,
Smith
C.
,
Youdale
T.
,
Leblanc
J.
,
Whitfield
J. F.
,
Sikorska
M.
(
1991
).
Topoisomerase II-reactive chemotherapeutic drugs induce apoptosis in thymocytes.
Cancer Res
51
,
1078
1085
Watts
D. J.
,
Ashworth
J. M.
(
1970
).
Growth of myxamoebae of the cellular slime mould Dictyostelium discoideum in axenic culture.
Biochem. J
119
,
171
174
Weismann
A.
(
1890
).
Prof. Weismann's theory of heredity.
Nature
41
,
317
323
Welker
D. L.
,
Hirth
K. P.
,
Williams
K. L.
(
1985
).
Inheritance of extrachromosomal ribosomal DNA during the asexual life cycle of Dictyostelium discoideum: examination by use of DNA polymorphisms.
Mol. Cell. Biol
5
,
273
280
White
K.
,
Grether
M. E.
,
Abrams
J. M.
,
Young
L.
,
Farrell
K.
,
Steller
H.
(
1994
).
Genetic control of programmed cell death in Drosophila.
Science
264
,
677
683
Whittingham
W. F.
,
Raper
K. B.
(
1960
).
Non-viability of stalk cells in Dictyostelium.
Proc. Nat. Acad. Sci. USA
46
,
642
649
Wyllie
A. H.
(
1980
).
Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation.
Nature
284
,
555
556
Wyllie
A. H.
,
Kerr
J. F. R.
,
Currie
A. R.
(
1980
).
Cell death: the significance of apoptosis.
Int. Rev. Cytol
68
,
251
306
Wyllie
A. H.
,
Morris
R. G.
,
Smith
A. L.
,
Dunlop
D.
(
1984
).
Chromatincleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis.
J. Pathol
142
,
67
77
Zakeri
Z. F.
,
Quaglino
D.
,
Latham
T.
,
Lockshin
R. A.
(
1993
).
Delayedinternucleosomal DNA fragmentation in programmed cell death.
FASEB J
7
,
470
478
This content is only available via PDF.