Summary
Embryos of the paedogenetically reproducing gall midge Heteropeza pygmaea develop floating in the haemocoel of a so-called mother larva. The egg membranes remain permeable and the embryos increase in size during embryonic development by taking up nutrients from the haemolymph. Such embryos can be cultured in vitro, i.e. in haemolymph drops obtained from mother larvae. We tested the effects of several drugs known to interact with cytoskeletal elements on different stages of embryonic development, including cleavage and gastrulation. The drugs were added to the in vitro cultures and the effects were studied with time-lapse cine-micrography. Colchicine and vinblastine blocked cleaving eggs in metaphase stage and arrested yolk globule oscillation. In spite of such a block blastoderms once formed continued development through germ band formation and extension and also increased in size. Cytochalasin B did not affect the stage of cleavage; however, it inhibited gastrulation and subsequent morphogenetic processes and also prevented size increase. We conclude that (1) the functioning of microtubules is needed for yolk globule oscillation during cleavage interphases but not for the gastrulation processes subsequent to blastoderm formation and (2) microfilaments do not play an important role in cleavage, at least not for the orderly succession of the cleavage divisions, but are essential for the morphogenetic movements associated with gastrulation. We suggest that during cleavage a limited stock of microtubules and their precursors is responsible for both transport of chromosomes during mitoses and translocation of organelles during interphase. Yolk oscillation seems to be a secondary effect and of minor or no importance for the normal course of embryonic development.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bajer AS, Molè-Bajer J (1982) Asters, poles, and transport properties within spindlelike microtubule arrays. Cold Spring Harbor Symp Quant Biol 46:263–283
Brown SS, Spudich JA (1981) Mechanism of action of cytochalasin: evidence that it binds to actin filament ends. J Cell Biol 88:487–491
Camenzind R (1962) Untersuchungen über die bisexuelle Fortpflanzung einer paedogenetischen Gallmücke. Rev Suisse Zool 69:377–384
De Brabander M, Geuens G, De Mey J, Joniau M (1981) Nucleated assembly of mitotic microtubules in living PTK2 cells after release from nocodazole treatment. Cell Motil 1:469–483
Dustin P (1984) Microtubules, 2nd edn. Springer, Berlin Heidelberg New York
Foe VE, Alberts BM (1983) Studies of nuclear and cytoplasmic behaviour during the five mitotic cycles that precede gastrulation in Drosophila embryogenesis. J Cell Sci 61:31–70
Fux T (1974) Chromosome elimination in Heteropeza pygmaea. II. Ultrastructure of the spindle apparatus. Chromosoma 49:99–112
Fux T, Camenzind R (1976) Effects of nitrous oxide on gall midge embryos: reversible arrest of mitosis and developmental disturbances. Cytobios 16:23–39
Fux T, Went DF, Camenzind R (1978) Movement pattern and ultrastructure of rotating follicles of the paedogenetic gall midge Heteropeza pygmaea Winnertz (Diptera: Cecidomyiidae). Int J Insect Morphol Embryol 7:415–426
Himes RH, Kersey RN, Heller-Bettinger I, Samson FE (1976) Action of the Vinca alkaloids vincristine, vinblastine, and desacetyl vinblastine amide on microtubules in vitro. Cancer Res 36:3798–3802
Junquera P (1983) The role of the follicular epithelium in growing eggs of a dipteran insect during late oogenesis and cleavage. J Morphol 178:303–312
Junquera P (1985) Cleavage and blastoderm formation in normal and experimentally deformed naked eggs of a dipteran insect. Wilhelm Roux's Arch 194:155–165
Kaiser J, Lang AB, Went DF (1982) Pulsation of nuclei in insect oocytes is reversibly inhibited by cytochalasin B. Exp Cell Res 139:460–463
Karr TL, Alberts BM (1986) Organization of the cytoskeleton in early Drosophila embryos. J Cell Biol 102:1494–1509
Miyamoto DM, van der Meer JM (1982) Early egg contractions and patterned parasynchronous cleavage in a living insect egg. Wilhelm Roux's Arch 191:95–102
Olmsted JB, Borisy GG (1973) Microtubules. Annu Rev Biochem 42:507–540
Plagemann PGW, Wohlhueter RM, Graff JC, Marz R (1978) Inhibition of carrier-mediated and non-mediated permeation processes by cytochalasin B. In: Tanenbaum SW (ed) Cytochalasins, biochemical and cell biological aspects. North-Holland Publishing Co., Amsterdam, pp 445–473
Rickoll WL (1976) Cytoplasmic continuity between embryonic cells and the primitive yolk sac during early gastrulation in Drosophila melanogaster. Dev Biol 49:304–310
Rickoll WL, Counce SJ (1980) Morphogenesis in the embryo of Drosophila melanogaster germ band extension. Wilhelm Roux's Arch 188:163–177
Sander K (1976) Morphogenetic movements in insect embryogenesis. In: Lawrence PA (ed) Insect development. Blackwell Scientific Publications, Oxford, pp 35–52
Schiff PB, Fant J, Horwitz SB (1979) Promotion of microtubule assembly in vitro by taxol. Nature 277:665–667
Spooner BS (1978) Cytochalasins as probes in selected morphogenetic processes. In: Tanenbaum SW (ed) Cytochalasins, biochemical and cell biological aspects. North-Holland Publishing Co., Amsterdam, pp 65–89
Stossel TP, Hartwig JH (1976) Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis. J Cell Biol 68:602–619
Suchard SJ, Goode D (1982) Microtubule-dependent transport of secretory granules during stalk secretion in a peritrich ciliate. Cell Motil 2:47–71
Takesue S, Owaribe K, Keino H (1984) Possible involvement of cytoskeletal organelles in blastoderm formation of the silkworm, Bombyx mori. XVIIth Int Congr Entomol Hamburg (GDR) 20–26 Aug 1984 (Abstr) S 8.2, p 419
Tucker JB, Meats M (1976) Microtubules and control of insect egg shape. J Cell Biol 71:207–217
Warn RM, Magrath R (1983) F-actin distribution during the cellularization of the Drosophila embryo visualized with FL-phalloidin. Exp Cell Res 143:103–114
Went DF (1971) In vitro culture of eggs and embryos of the viviparous paedogenetic gallmidge Heteropeza pygmaea. J Exp Zool 177:301–312
Went DF (1972) Zeitrafferfilmanalyse der Embryonalentwicklung in vitro der vivipar paedogenetischen Gallmücke Heteropeza pygmaea. Wilhelm Roux's Arch 170:13–47
Went DF (1977) In vitro culture of ovaries of a viviparous gall midge. In Vitro 13:76–84
Went DF (1978) Ovarentwicklung und Follikelbildung bei der Gallmücke Heteropeza pygmaea. Tonfilm D 1267, Inst Wiss Film Göttingen, Publ Wiss Film Sekt Biol 11/26:1–10
Went DF (1982) Insect ovaries and follicles in culture: oocyte and early embryonic development in pedogenetic gall midges. Adv Cell Cult 2:197–235
Went DF, Junquera P (1981) Embryonic development of insect eggs formed without follicular epithelium. Dev Biol 86:100–110
Went DF, Fux T, Camenzind R (1978) Movement pattern and ultrastructure of pulsating oocyte nuclei of the paedogenetic gall midge, Heteropeza pygmaea Winnertz (Diptera: Cecidomyiidae). Int J Insect Morphol Embryol 7:301–314
Wessels NK, Spooner BS, Ash JF, Bradley MO, Luduena MA, Taylor EL, Wrenn JT, Yamada KM (1971) Microfilaments in cellular and developmental processes. Science 171:135–143
Wilson L, Anderson K, Creswell K (1974) On the mechanism of action of vinblastine. J Cell Biol 63:373a
Wilson L, Creswell KM, Chin D (1975) The mechanism of action of vinblastine. Binding of [acetyl-3H]vinblastine to embryonic chick brain tubulin and tubulin from sea urchin sperm tail outer doublet microtubules. Biochemistry 14:5586–5592
Wolf R (1978) The cytaster, a colchicine-sensitive migration organelle of cleavage nuclei in an insect egg. Dev Biol 62:464–472
Wolf R (1980) Migration and division of cleavage nuclei in the gall midge, Wachtliella persicariae. II. Origin and ultrastructure of the migration cytaster. Wilhelm Roux's Arch 188:65–73
Wolf R (1985) Migration and division of cleavage nuclei in the gall midge, Wachtliella persicariae. III. Pattern of anaphase-triggering waves altered by temperature gradients and local gas exchange. Wilhelm Roux's Arch 194:257–270
Zalokar M, Erk I (1976) Division and migration of nuclei during early embryogenesis of Drosophila melanogaster. J Microsc Biol Cell 25:97–106
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Gerhard Krause on the occasion of his 80th birthday
Rights and permissions
About this article
Cite this article
Kaiser, J., Went, D.F. Early embryonic development of the dipteran insect Heteropeza pygmaea in the presence of cytoskeleton-affecting drugs. Roux's Arch Dev Biol 196, 356–366 (1987). https://doi.org/10.1007/BF00375772
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00375772