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Pellicular changes during division in Astasia longa
Pellicular
changes during
division
in Astasia longa
REFERENCES
1. 2.
BARER,
__
R.,
Phys.
Nature Techn.
169,
336
Biol.
Res. 3, 30 (1956).
(1952).
R. and JOSEPH, S., Quart. J. Micr. Sci. 96, 423 (1955 ix). R., and TKACZYK, S., Nafure 173, 821 (1954). H. G., General Cytochemical Methods, J. F. DANIELLI (ed.) vol. 1, p. 55. Academic Press, New York 1958. Davms, H. G., WILKINS, M. H. F., CIIAYEN, J. and LA COUR, L. F., Quarf. J. Micr. Sci. 95, 271 (1954). DAVIES, k. G. bd DEELEY, E. M., Exptl CeZl Res. 11, 169 (1956). DAVIES. 111. G. and WILKINS. M. H. F.. Report to the Cvtochemistrv Commission of the Sdciety for Cell Biology; Physical AspeEts of Cytochemical Metho”ds, Stockholm, 1951. __ Nature 169, 541 (1952). HALE, A. J., The Interference Microscope in Biological Research. E. & S. LIVINGSTONE Ltd. Edinburgh and London, 1958. HUXLEY, H. E. and HANSON, J., Biochim. Biophys. Acta 23, 229 (1957). HYDISN, H., Biochemistry of the Central Nervous System, BRUCKE F. (ed.) vol. 3, 61. Pergamon Press, New York, 1958. MITCHISON, J. M., PASSANO, L. M. and SMITH, F. H., Quarf. J. Micr. Sci. 97, 287 (1956). PELLEGRIN~, C. and TONGIANI, R., Riu. Isfoch. norm. iat: 53 (1962). SVENSSON, G., Ezpfl Cell Res. 12, 406 (1957). VIOLA, M. P. and PUCCINELLI, E., Riu. Istoch. norm. pat. In Press.
3. BARER, 4. BARER, 5. DAVIES, G. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.
PELLICULAR IN
CHANGES
DURING
ASTASIA
LONG/d
J. R. SOMMER Departments
of Pathology
and J. J. BLUM
and Physiology, Durham, N.C., Received
March
DIVISION
Duke U.S.A. 31,
University
Medical
Center,
19641
Synchronized cultures constitute a source of large numbers of cells at various stages of division, and are therefore ideally suited for the study of cell division at the fine structure level. Light microscopic observations of the nuclear changes of Asfasia longa, synchronized by means of a repetitive temperature cycle [I], have shown that most of the population is in late prophase about 2 hr after the beginning of the warm period. Cells were collected from such cultures by brief centrifugation, fixed in glutaraldehyde for 24 hr, post-fixed in osmium tetroxide, embedded in Maraglas, sectioned with an ultramicrotome, and viewed with the electron microscope. The details of the structure of the pellicle complex of Euglena gracilis have recently been analysed [4]. Each pellicle complex contains one tubule of the endoplasmic reticulum and four 1 Revised 28-
641804
version
June
22,
1964.
Experimental
Cell
Research
35
J. R. Sommer and J. J. Blum tubular fibrils characteristically located with respect to the notch. Astasia longa, a close relative of Eugkna gracilis, has virtually the same pellicle complex organization, except that the notch appears to be more prominent (Fig. 1). Cells taken in early prophase show that each pellicle complex displays a presumptive ridge originating next to the adult ridge (Fig. 2). The size of the presumptive ridge increases as division proceeds (Fig. 3), and becomes comparable to the adult ridges at an as yet undetermined stage of cytokinesis (Fig. 4). It should be noted that the presumptive ridges already possess the characteristic four tubular fibrils (Figs. 3, 5). This is also observed in dividing Euglena gmcilis. Presumptive pellicle complexes are also prominent in the cytostome of non-dividing Euglena gracilis [4, 51. The duplication of pellicle complexes is an essential feature of cytokinesis in the Eugleninae [2, 31. The duplication of pellicle complexes is a process of intercalary growth but presumably begins at the gullet-cytostome junction during early prophase, and is probably preceded by duplication of tubular fibrils. The duplication does not seem to be effected by bifurcation. While such bifurcations (or fusions), exist at the posterior vortex of the cell, we have not observed them in the cytostome region where fission begins in the Eugleninae. This work was supported by grant numbers GM-09730 and CA-06911 from the National Institutes of Health. One of us (J. .J. Blum) acknowledges the support of a Public Health Service Career Program Award (#l-K3-GM-2341) from the National Institute of General Medical Sciences. The authors are indebted to Mr. N. Sarji and Miss Susan Kimball for their excellent technical assistance.
REFERENCES 1. 2. 3. 4. 5.
BLUM, J. J. and PADILLA, G. If., Bxpti Cell Res. 28, 512 (1962). JAMES, T. W., Sgmp. Int. Sac. Cell Bid. 2, (1962). POCHMANV, A., Planta 42, 478 (1963). SOMMER, J. R., I. Cell Dial. In Press. SOMMEH, J. R. and BLUM, J. J., I. Cell Biol. In Press.
Fig. l.-Pellicle subpellicular x 44,000.
of Astasia longa endoplasmic reticulum
Fig. 2.-Pelliclc of Astasia ridges (BR). Golgi complex
in resting (SER),
phase. Cell membrane notch (N), tubular fibril
longa in early prophase. at (C). x 40,000.
Note
adult
(CM), (TF), ridges
ridge (R), groove (G), mitochondrium (MIT). (AR)
with
Fig. 3.-Pellicle of Astasia longa in late prophase. Note (BR). Adult ridge (AR); tubular fibril (TF). x 40,000.
increase
Fig. 4.-Pellicle of Astasia presumptive ridges (BR),
the extent of the alternating cells. x 19,200.
longa in late prophase and adult ridges (AR),
Fig. 5.-Pellicle of Euglena gracilis in division. elevated presumptive ridges. x 40,000.
Experimental
Cell
Research
35
showing in two Note
tubular
fibrils
presumptive
in size of presumptive
(TF)
underlying
ridges pattern
as yet
barely
of
Pellicular
changes during
division
in Astasia longa
425
changes during
division
in Astasia longa
REFERENCES
1. 2.
BARER,
__
R.,
Phys.
Nature Techn.
169,
336
Biol.
Res. 3, 30 (1956).
(1952).
R. and JOSEPH, S., Quart. J. Micr. Sci. 96, 423 (1955 ix). R., and TKACZYK, S., Nafure 173, 821 (1954). H. G., General Cytochemical Methods, J. F. DANIELLI (ed.) vol. 1, p. 55. Academic Press, New York 1958. Davms, H. G., WILKINS, M. H. F., CIIAYEN, J. and LA COUR, L. F., Quarf. J. Micr. Sci. 95, 271 (1954). DAVIES, k. G. bd DEELEY, E. M., Exptl CeZl Res. 11, 169 (1956). DAVIES. 111. G. and WILKINS. M. H. F.. Report to the Cvtochemistrv Commission of the Sdciety for Cell Biology; Physical AspeEts of Cytochemical Metho”ds, Stockholm, 1951. __ Nature 169, 541 (1952). HALE, A. J., The Interference Microscope in Biological Research. E. & S. LIVINGSTONE Ltd. Edinburgh and London, 1958. HUXLEY, H. E. and HANSON, J., Biochim. Biophys. Acta 23, 229 (1957). HYDISN, H., Biochemistry of the Central Nervous System, BRUCKE F. (ed.) vol. 3, 61. Pergamon Press, New York, 1958. MITCHISON, J. M., PASSANO, L. M. and SMITH, F. H., Quarf. J. Micr. Sci. 97, 287 (1956). PELLEGRIN~, C. and TONGIANI, R., Riu. Isfoch. norm. iat: 53 (1962). SVENSSON, G., Ezpfl Cell Res. 12, 406 (1957). VIOLA, M. P. and PUCCINELLI, E., Riu. Istoch. norm. pat. In Press.
3. BARER, 4. BARER, 5. DAVIES, G. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16.
PELLICULAR IN
CHANGES
DURING
ASTASIA
LONG/d
J. R. SOMMER Departments
of Pathology
and J. J. BLUM
and Physiology, Durham, N.C., Received
March
DIVISION
Duke U.S.A. 31,
University
Medical
Center,
19641
Synchronized cultures constitute a source of large numbers of cells at various stages of division, and are therefore ideally suited for the study of cell division at the fine structure level. Light microscopic observations of the nuclear changes of Asfasia longa, synchronized by means of a repetitive temperature cycle [I], have shown that most of the population is in late prophase about 2 hr after the beginning of the warm period. Cells were collected from such cultures by brief centrifugation, fixed in glutaraldehyde for 24 hr, post-fixed in osmium tetroxide, embedded in Maraglas, sectioned with an ultramicrotome, and viewed with the electron microscope. The details of the structure of the pellicle complex of Euglena gracilis have recently been analysed [4]. Each pellicle complex contains one tubule of the endoplasmic reticulum and four 1 Revised 28-
641804
version
June
22,
1964.
Experimental
Cell
Research
35
J. R. Sommer and J. J. Blum tubular fibrils characteristically located with respect to the notch. Astasia longa, a close relative of Eugkna gracilis, has virtually the same pellicle complex organization, except that the notch appears to be more prominent (Fig. 1). Cells taken in early prophase show that each pellicle complex displays a presumptive ridge originating next to the adult ridge (Fig. 2). The size of the presumptive ridge increases as division proceeds (Fig. 3), and becomes comparable to the adult ridges at an as yet undetermined stage of cytokinesis (Fig. 4). It should be noted that the presumptive ridges already possess the characteristic four tubular fibrils (Figs. 3, 5). This is also observed in dividing Euglena gmcilis. Presumptive pellicle complexes are also prominent in the cytostome of non-dividing Euglena gracilis [4, 51. The duplication of pellicle complexes is an essential feature of cytokinesis in the Eugleninae [2, 31. The duplication of pellicle complexes is a process of intercalary growth but presumably begins at the gullet-cytostome junction during early prophase, and is probably preceded by duplication of tubular fibrils. The duplication does not seem to be effected by bifurcation. While such bifurcations (or fusions), exist at the posterior vortex of the cell, we have not observed them in the cytostome region where fission begins in the Eugleninae. This work was supported by grant numbers GM-09730 and CA-06911 from the National Institutes of Health. One of us (J. .J. Blum) acknowledges the support of a Public Health Service Career Program Award (#l-K3-GM-2341) from the National Institute of General Medical Sciences. The authors are indebted to Mr. N. Sarji and Miss Susan Kimball for their excellent technical assistance.
REFERENCES 1. 2. 3. 4. 5.
BLUM, J. J. and PADILLA, G. If., Bxpti Cell Res. 28, 512 (1962). JAMES, T. W., Sgmp. Int. Sac. Cell Bid. 2, (1962). POCHMANV, A., Planta 42, 478 (1963). SOMMER, J. R., I. Cell Dial. In Press. SOMMEH, J. R. and BLUM, J. J., I. Cell Biol. In Press.
Fig. l.-Pellicle subpellicular x 44,000.
of Astasia longa endoplasmic reticulum
Fig. 2.-Pelliclc of Astasia ridges (BR). Golgi complex
in resting (SER),
phase. Cell membrane notch (N), tubular fibril
longa in early prophase. at (C). x 40,000.
Note
adult
(CM), (TF), ridges
ridge (R), groove (G), mitochondrium (MIT). (AR)
with
Fig. 3.-Pellicle of Astasia longa in late prophase. Note (BR). Adult ridge (AR); tubular fibril (TF). x 40,000.
increase
Fig. 4.-Pellicle of Astasia presumptive ridges (BR),
the extent of the alternating cells. x 19,200.
longa in late prophase and adult ridges (AR),
Fig. 5.-Pellicle of Euglena gracilis in division. elevated presumptive ridges. x 40,000.
Experimental
Cell
Research
35
showing in two Note
tubular
fibrils
presumptive
in size of presumptive
(TF)
underlying
ridges pattern
as yet
barely
of
Pellicular
changes during
division
in Astasia longa
425