The role of microtubules in contractile ring function.

During cytokinesis, a cortical contractile ring forms around a cell, constricts to a stable tight neck and terminates in separation of the daughter cells. At first cleavage, Ilyanassa obsoleta embryos form two contractile rings simultaneously. The cleavage furrow (CF), in the animal hemisphere between the spindle poles, constricts to a stable tight neck and separates the daughter cells. The third polar lobe constriction (PLC-3), in the vegetal hemisphere below the spindle, constricts to a transient tight neck, but then relaxes, allowing the polar lobe cytoplasm to merge with one daughter cell. Eggs exposed to taxol, a drug that stabilizes microtubules, before the CF or the PLC-3 develop, fail to form CFs, but form stabilized tight PLCs. Eggs exposed to taxol at the time of PLC-3 formation develop varied numbers of constriction rings in their animal hemispheres and one PLC in their vegetal hemisphere, none of which relax. Eggs exposed to taxol after PLC-3 initiation form stabilized tight CFs and PLCs. At maximum constriction, control embryos display immunolocalization of nonextractable alpha-tubulin in their CFs, but not in their PLCs, and reveal, via electron microscopy, many microtubules extending through their CFs, but not through their PLCs. Embryos which form stabilized tightly constricted CFs and PLCs in the presence of taxol display immunolocalization of nonextractable alpha-tubulin in both constrictions and show many polymerized microtubules extending through both CFs and PLCs. These results suggest that the extension of microtubules through a tight contractile ring may be important for stabilizing that constriction and facilitating subsequent cytokinesis.     

Control of Polar Lobe Formation in Fertilized Eggs of Ilyanassa obsoleta Stimpson

Mechanisms of polar lobe formation in fertilized eggs of Ilyanassaobsoleta are discussed. Data are presented in the context ofa model involving a contractile ring of microfilaments in thecortical cytoplasm of the vegetal hemisphere. The polar lobeneck diameter decreases at two distinct rates during formationof second, third, and fourth polar lobes. During the second,more rapid phase of lobe constriction, eggs contain a band ofmicrofilaments arranged circumferentially in the cortical cytoplasmapposed to the plasma membrane at the base of the polar lobeconstriction. These microfilaments disappear and lobe constrictionsregress in cytochalasin B, but not in colchicine. Colchicineprevents eggs from beginning the second, more rapid phase oflobe constriction. Eggs require the presence of potassium ionsbut not sodium ions for normal polar lobe formation and cleavage.When eggs are placed in isotonic solutions of CaCl₂, within10–15 min they form cytoplasmic blebs which enlarge intolobes. This calcium-induced blebbing is inhibited by cytochalasinB but not by colchicine. Blebbing occurs in the calcium concentrationrange of 0.17 M-0.34 M in the presence of NaCl, MgCl₂, or MnCl₂.Potassium ions actively inhibit the calcium-induced blebbing,however.     

Effects of local anesthetics on cytokinesis and polar lobe formation in fertilized eggs of Ilyanassa obsoleta

Summary

We have examined the ability of fertilized eggs of Ilyanassa obsoleta to form polar lobe constrictions and undergo cytokinesis in the presence of several local anesthetics and compared these effects with those of drugs known to affect microtubules. Procaine, lidocaine (Xylocaine), mepivacaine, tetracaine, and dibucaine all delay the beginning of polar lobe constrictions at low concentrations and in the order of their lipid solubilities. All of the anesthetics are effective at lower concentrations in the absence of extracellular Ca²⁺. Procaine and tetracaine ‘lock’ cells for several hours halfway through the constriction of the polar lobe neck and prevent subsequent cytokinesis, effects similar to those of the microtubule agents, colchicine and nocodazole. Procaine has no effect on membrane potential, ψ_m, or on intracellular chloride activity, (Cl)_c, as determined with ion-selective microelectrodes. This suggests that procaine does not inhibit cellular shape changes by affecting the ionic activities of the predominant intracellular cation (K⁺) or anion (Cl^?).     

Polar lobe formation and cytokinesis in fertilized eggs of Ilyanassa obsoleta. II. Large bleb formation caused by high concentrations of exogenous calcium ions

Fertilized eggs of the mollusk Ilyanassa obsoleta (Nassarius obsoletus) form large blebs resembling polar lobes within 12 min of exposure to solutions of isotonic CaCl₂, whereas control eggs in sea water remain spherical. Under identical conditions, fertilized eggs of the sea urchin, Strongylocentrotus purpuratus, an organism which normally does not form polar lobes, do not form blebs upon exposure to solutions of isotonic CaCl₂. The calcium-induced blebbing in Ilyanassa still occurs if other cations such as Na⁺, Mg²⁺, or Mn²⁺ are present in addition to Ca²⁺, but not if comparable concentrations of K⁺ are present. Cytochalasin B prevents the calcium-induced blebbing, whereas colchicine does not. Cytokinesis in both Ilyanassa and Strongylocentrotus and normal polar lobe formation in Ilyanassa appear to require exogenous K⁺ but not exogenous Ca²⁺. Preliminary electron microscopy of Ilyanassa eggs exposed to isotonic solutions of CaCl₂ has shown microfilaments in the cortical cytoplasm in the region of the bleb constriction but no microfilaments in spherical control eggs in sea water. These data suggest that high concentrations of exogenous Ca²⁺ trigger the polymerization and contraction of a ring of microfilaments in the cortical cytoplasm of the Ilyanassa egg which results in the formation of a lobelike bleb of cytoplasm. The observation that K⁺ antagonizes this Ca²⁺-induced blebbing has led to the formulation of a theory which postulates that the ratio of intracellular Ca²⁺ to intracellular K⁺ is critical in the control of polar lobe formation and cytokinesis.     

Localization of caldesmon and its dephosphorylation during cell division

Mitosis-specific phosphorylation by cdc2 kinase causes nonmuscle caldesmon to dissociate from microfilaments during prometaphase. (Yamashiro, S., Y. Yamakita, R. Ishikawa, and F. Matsumura. 1990. Nature (Lond.). 344:675-678; Yamashiro, S., Y. Yamakita, H. Hosoya, and F. Matsumura. 1991. Nature (Lond.) 349:169-172). To explore the functions of caldesmon phosphorylation during cytokinesis, we have examined the relationship between the phosphorylation level, actin- binding, and in vivo localization of caldesmon in cultured cells after their release of metaphase arrest. Immunofluorescence studies have revealed that caldesmon is localized diffusely throughout cytoplasm in metaphase. During early stages of cytokinesis, caldesmon is still diffusely present and not concentrated in contractile rings, in contrast to the accumulation of actin in cleavage furrows during cytokinesis. In later stages of cytokinesis, most caldesmon is observed to be yet diffusely localized although some concentration of caldesmon is observed in cortexes as well as in cleavage furrows. When daughter cells begin to spread, caldesmon shows complete colocalization with F- actin-containing structures. These observations are consistent with changes in the levels of microfilament-associated caldesmon during synchronized cell division. Caldesmon is missing from microfilaments in prometaphase cells arrested by nocodazole treatment, as shown previously (Yamashiro, S., Y. Yamakita, R. Iskikawa, and F. Matsumura. 1990. Nature (Lond.). 344:675-678). The level of microfilament- associated caldesmon stays low (12% of that of interphase cells) when some cells start cytokinesis at 40 min after the release of metaphase arrest. When 60% of cells finish cytokinesis at 60 min, the level of microfilament-associated caldesmon is recovered to 50% of that of interphase cells. The level of microfilament-associated caldesmon is then gradually increased to 80% when cells show spreading at 120 min. Dephosphorylation appears to occur during cytokinesis. It starts when cells begin to show cytokinesis at 40 min and completes when most cells finish cytokinesis at 60 min. These results suggest that caldesmon is not associated with microfilaments of cleavage furrows at least in initial stages of cytokinesis and that dephosphorylation of caldesmon appears to couple with its reassociation with microfilaments. Because caldesmon is known to inhibit actomyosin ATPase and/or regulate actin assembly, its continued dissociation from microfilaments may be required for the assembly and/or activation of contractile rings.     

Holoblastic early cleavage of Tetrodontophora bielanensis (Collembola) eggs, with special reference to its irregularity.

The fertilized eggs of Tetrodontophora bielanensis start to cleave 6 to 8 days after oviposition and initially only karyokineses occur. The cytokinesis begins after two karyokineses, when four nuclei are observed in the ooplasm. Two cleavage furrows, perpendicular to each other, appear simultaneously at the egg poles where polar bodies are located and gradually the furrows encompass the whole egg diameter. The furrow formation is initiated by the bundle of microfilaments that contract and pull superficial fragments of the oolemma into the yolk and subsequently new membranes, separating the daughter cells, start to form. However, they do not grow towards the egg centre but bifurcate, leaving the central part of the ooplasm outside of the newly formed blastomeres. Starting from the fourth or fifth cleavage division, the bifurcations permanently occur and multiple cleavage furrows are formed on the embryo surface. Moreover, fragments of the ooplasm, enclosed within the cell membrane but devoid of cell nucleus are observed. During further development such cell fragments become reincorporated into the embryo. This mode of cleavage leads eventually to the formation of cellular blastoderm on the embryo surface. The results presented in the paper suggest that the control of cleavage in T. bielanensis acts not at the level of cytoplasmic determinants but rather at the level of positional information of blastomeres.     

Effect of hexylene glycol-altered microtubule distributions on cytokinesis and polar lobe formation in fertilized eggs of Ilyanassa obsoleta

Some effects of gravity on early morphogenesis are correlated with microtubule locations within cells. During first cleavage in Ilyanassa obsoleta embryos, a transitory polar lobe constriction forms and then relaxes, allowing the polar lobe to merge with one daughter cell. If the polar lobe is equally divided or removed, morphogenesis is severely disrupted. To examine microtuble locations during early Ilyanassa development, eggs were fixed and stained for polymerized alpha-tubulin during first cleavage. The mitotic apparatus assembles at the animal pole. The cleavage furrow forms between the asters, constricting to a stabilized intercellular bridge encircling midbody-bound microtubules, whereas the polar lobe constriction forms below and parallel to the spindle, constricting to a transitory intercellular bridge encircling no detectable microtubules. At metaphase an alpha-tubulin epitope is distributed throughout the spindle, whereas a beta-tubulin epitope is present predominantly in the asters. Incubation in hexylene glycol, a drug that increases microtubule polymerization, during mitosis causes the polar lobe constriction to tighten around polymerized alpha-tubulin and remain stably constricted. If hexylene glycol is removed, alpha-tubulin staining disappears from the polar lobe constriction, which relaxes, whereas microtubules remain in the cleavage furrow, which remains constricted. These observations suggest that asymmetric distribution of microtubules affects early Ilyanassa cleavage patterns, and that continued presence of microtubules extending through an intercellular bridge is important for stabilization of the bridge constriction prior to completion of cytokinesis. These data provide the basis for further analysis of the role of microtubules in possible microgravity disruptions of Ilyanassa development.     

Polar organizers and girdling bands of microtubules are associated with γ-tubulin and act in establishment of meiotic quadripolarity in the hepatic Aneura pinguis (Bryophyta)

Summary. Meiosis in Aneura pinguis is preceded by extensive cytoplasmic preparation for quadripartitioning of the diploid sporocyte into a tetrad of haploid spores. In early prophase the four future spore domains are defined by lobing of the cytoplasm and development of a quadripolar prophase spindle focused at polar organizers (POs) centered in the lobes. Cells entering the reproductive phase become isolated and, instead of hooplike cortical microtubules, have endoplasmic microtubule systems centered on POs. These archesporial cells proliferate by mitosis before entering meiosis. In prophase of each mitosis, POs containing a distinct concentration of γ-tubulin appear de novo at tips of nuclei and initiate the bipolar spindle. Cells entering meiosis become transformed into quadrilobed sporocytes with four POs, one in each lobe. This transition is a complex process encompassing assembly of two opposite POs which subsequently disperse into intersecting bands of microtubules that form around the central nucleus. The girdling bands define the future planes of cytokinesis and the cytoplasm protrudes through the restrictive bands becoming quadrilobed. Two large POs reappear in opposite cleavage furrows. Each divides and the resulting POs migrate into the tetrahedral lobes of cytoplasm. Cones of microtubules emanating from the four POs interact to form a quadripolar microtubule system (QMS) that surrounds the nucleus in meiotic prophase. The QMS is subsequently transformed into a functionally bipolar metaphase spindle by migration of poles in pairs to opposite cleavage furrows. These findings contribute to knowledge of microtubule organization and the role of microtubules in spatial regulation of cytokinesis in plants. Correspondence and reprints: Department of Biology, University of Louisiana-Lafayette, Lafayette, LA 70504-2451, U.S.A.     

Polar effects of concanavalin A on the cortical cytoskeleton of a molluscan egg (Nassarius reticulatus,Gastropoda)

Summary The organization of the surface of fertilizedNassarius reticulatus eggs was probed by investigating the effects of treatment with concanavalin A (Con A). This lectin causes abnormal polar lobe formation as well as inhibition of cleavage. At low concentrations of Con A (0.3–1.0 μg/ml) the polar lobe constriction becomes considerably elongated, whereas at higher concentrations (2.5–50 μ/ml) the position of the constriction undergoes an extreme shift towards the animal pole. In the latter case, the surface of the animal part of the egg forms large protrusions and folds. Con A also causes resorption of microvilli and disappearance of the extracellular layer covering these villi; this process starts at the vegetal pole and propagates towards the animal pole. These changes in surface architecture are associated with profound changes in the organization of filamentous (F-) actin as assessed by confocal laser scanning microscopy of NBD-phallacidin-labelled eggs. Divalent succinyl-Con A has the same effects on polar lobe formation and surface architecture as tetravalent Con A, but only at very high concentrations (100–200 μg/ml), indicating that Con A exerts its effects by cross-linkage of its binding sites. Experiments with cytoskeleton inhibitors (cytochalasin D, colchicine, and nocodazole) reveal that in Con A-treated eggs — as in untreated eggs — microfilaments, but not microtubules, are involved in the formation of the polar lobe constriction. The calcium ion channel blocker D600 affects neither normal nor Con A-induced abnormal polar lobe formation, which suggests that influx of external calcium is not required. In contrast, treatment with TMB-8, an antagonist of internal calcium release, prevents the formation of a polar lobe in both normal and Con A-treated eggs. Finally, eggs from which the polar lobe has been removed prior to Con A treatment show none of the effects described, whereas isolated polar lobes react similarly to intact eggs. These results suggest that binding of Con A to sites present at the vegetal pole of the egg is responsible for the observed effects of the lectin.     

Effect of microinjected N-ethylmaleimide-modified heavy meromyosin on cell division in amphibian eggs

N-Ethylmaleimide-modified heavy meromyosin (NEM-HMM) microinjected into amphibian eggs inhibits cytokinesis and the cortical contractions associated with wound closure. Injection of NEM-HMM into two-cell Rana pipiens embryos produces a zone of cleavage inhibition around the point of injection. Early furrows followed by time-lapse microcinematography are seen to slow and stop as they enter the NEM-HMM-injected zone. Arrested furrows slowly regress, leaving a large region of cytoplasm uncleaved. Few nuclei are found in these regions of cleavage inhibition. Wound closure is often inhibited by NEM-HMM, especially when this inhibitor is injected just beneath the egg cortex. We observe that the surface of an unfertilized Rana egg is covered with microvilli that disappear during the course of development. The surfaces of NEM- HMM-inhibited zones remain covered with microvilli and resemble the unfertilized egg surface.     

Predetermination of cytokinesis and transition from holoblastic to partial superficial cleavage in early embryonic development of Tetrodontophora bielanensis (Collembola)

In the T. bielanensis embryo, only karyokinesis occurs during the first cleavage division, and a two-nuclear syncytial embryo forms. Then, two cytoplasmic concentrations in the form of elongated rolls perpendicular to each other develop below the periplasm at the animal pole of the egg. The second cleavage division is also associated with karyokineses only. After the embryo reaches the four-nuclear stage, cytokinesis occur at its animal pole, and two cleavage furrows perpendicular to each other develop in the periplasm above the cytoplasmic concentrations. The cell membranes forming within the furrows do not invade the cytoplasmic concentrations, but their growing tips push them into the egg interior, where they merge and form the central cytoplasmic concentration. The developing cell membranes do not invade the central cytoplasm; they band and grow above its surface. Four pyramidal blastomeres form as a result of this. The eight-blastomere embryo forms through both karyokinesis and cytokinesis, but the growing cell membranes now band below the previous ones and cut off anucleate parts of the mother blastomeres, which fuse with the central cytoplasm. Thus, during this phase of development the transition from holoblastic to partial superficial cleavage is initiated. Morphological analysis suggests that the formation of the first two cytokinesis is predetermined by and depends on factors connected with the animal pole periplasm. It also suggests that the central cytoplasm constitutes the morphological field, inducing the transition from holoblastic to partial superficial cleavage.     

The ultrastructural organization of the isolated cortex in eggs ofNassarius reticulatus (Mollusca)

Summary We have studied the organization of the cortex in fertilized eggs ofNassarius reticulatus by examining rotary-shadowed whole mounts of isolated cortices in the transmission electron microscope. The following components were distinguished: (a) the plasma membrane, with clathrin-coated areas and coated pits, (b) microfilaments and microtubules, and (c) a tubulovesicular network of endoplasmic reticulum. Microfilaments were identified by labeling with heavy meromyosin, and microtubules with a monoclonal anti-tubulin antibody, using both immunofluorescence microscopy and immunogold labeling for transmission electron microscopy. The microfilaments are organized in a network parallel to and closely associated with the plasma membrane, with typical Y- and X-shaped intersections. The endoplasmic reticulum is associated with this microfilamentous lattice. The microtubules also run parallel to the plasma membrane, but they are located at a greater distance, as can be inferred from stereo images. In the uncleaved egg, numerous microtubules are present in the egg cortex. Shortly before polar lobe formation, at the onset of mitosis, the microtubules disappear almost entirely. They reappear again at the end of first cleavage, as the polar lobe is being resorbed. The synthesis of cortical microtubules at this stage appears to depend on the presence of microtubule-organizing centers in the animal hemisphere of the egg, since microtubules do not reappear in isolated polar lobes. Clathrin-coated areas are present in both the animal and vegetal hemisphere before polar lobe formation. During mitosis, the clathrin-coated plaques and pits are found almost exclusively in the animal hemisphere. After resorption of the polar lobe, at the two-cell stage, no clathrin-coated areas were found at all.     

Cell Division in Tetraedron

Mitosis and cytokinesis in Tetraedron are described. Persistentcentrioles replicate before division and the pairs separateto define the future poles of the spindle whilst increasingnumbers of microtubules become associated with them. By prophase,the centrioles and most extranuclear microtubules have becomeenclosed within a 'perinuclear envelope' of endoplasmic reticulum.The nuclear envelope near the centrioles then becomes indentedand finally ruptures to form polar fenestrae during prometaphase;the extranuclear microtubules soon vanish and appear to movethrough the fenestrae into the forming spindle. Metaphase, anaphase,and telophase follow as usual. After mitosis, arrays of 'phycoplast'microtubules proliferate between nuclei. The cytoplasm is cleavedby membrane furrows coplanar with and growing through the phycoplasttubules. However, this cleavage is delayed until the cells havebecome multinucleate, and it appears to be irregular in extentand disposition in the cell until after a final set of synchronousmitoses. Then cytokinesis cuts up the cytoplasm into numeroussmall autospores which secrete their own wall; they are laterreleased following rupture of the parental wall. Some autosporesare binucleate which indicates that this cleavage apparatusdoes not necessarily cut up all the cytoplasm into uninucleatesegments. Vegetative reproduction in these organisms is comparedto that of other members of the Chlorococcales.     

Visualization of actin polymerization and depolymerization cycles during polyamine-induced cytokinesis in living Amoeba proteus

Summary Microinjection of spermine induces cytokinesis of Amoeba proteus. Within 30–60 s after spermine injection cells form one, or less commonly, two cleavage furrows and within the following 4–10 min the constrictions are completed. The resulting nucleated cell parts show normal streaming and locomotion, whereas the non-nucleated cell parts remain stationary and later degenerate.The intracellular distribution of fully polymerization-competent fluorescently labelled muscle actin was followed by image intensification. Double injection experiments initially using labelled actin and 30 min later spermine revealed a ring-like structure of enhanced fluorescence corresponding to the constricting cleavage furrow. Immediately after cleavage was completed, the ring disappeared. Electron microscopy of cells fixed during spermine-induced cytokinesis showed numerous well aligned actin and myosin filaments in the developing cleavage furrow. These filaments are a specialized manifestation of the cell cortex.The results demonstrate that cycles of actin and myosin polymerization and depolymerization and the parallel alignment of preexisting filaments (crosslinking) represent a basic mechanism in the generation of the motive force during cytokinesis.     

Effect of sodium dodecyl sulfate on polar lobe formation and function in Ilyanassa obsoleta embryos

Polar lobes, anucleate vegetal pole protrusions formed by Ilyanassa obsoleta embryos, serve as a mechanism for shunting morphogenetic determinants to one cell during the first two cleavages. Polar lobe material becomes segregated in the CD cell during first cleavage and in the D cell during second cleavage, resulting in a very unequal four-cell stage. Larval structures including external shell, foot, operculum, statocysts, and eyes develop only when polar lobe material is present. Treatment with the anionic detergent sodium dodecyl sulfate (SDS) before and during the first cleavage inhibited polar lobe formation and equalized cleavage, as the lobe material was distributed to two cells. No polar lobes formed during second clevage in SDS-equalized embryos, and the four-cell stage consisted of four equal cells with reduced cell contacts. SDS inrreversibly inhibited polar lobe formation without affecting cytokinesis. Although 27% of the larvae from SDS-equalized embryos had one or more lobe-dependent structures duplicated, morphogenesis was impaired: more than 40% of such larvae failed to form shell and/or statocysts. When cells were separated after equalized first cleavage and raised as pairs, the pairs of resulting larvae duplicated lobe-dependent structures with the same frequency as whole equalized embryos. Possible explanations for impaired morphogenesis in SDS-treated embryos are discussed.     

Myosin and actin are necessary for polar lobe formation and resorption in Ilyanassa obsoleta embryos

 During the first mitotic divisions many spiralian embryos form a cytoplasmic protrusion at the vegetal pole called the polar lobe. In the gastropod Ilyanassa obsoleta the polar lobe is constricted by a contractile ring composed of filamentous actin, myosin, and associated proteins, similar to the contractile ring of the cleavage furrow. To resolve the role of myosin and actin in polar lobe formation and resorption, we have applied 2,3-butanedione monoxime and Latrunculin B at different stages of the first cleavage to inhibit myosin and F-actin, respectively. Our results show that myosin is important for both cytokinesis and polar lobe formation. Additionally, we have found that the resorption of the polar lobe is a two-step process: the first step is passive, driven by the tension of the actin-cortex and the second step is active, in which the ATP-hydrolysis of myosin/actin interaction supplies the force to complete the resorption of the polar lobe. We have summarized our results in a scheme of the first cleavage of Ilyanassa obsoleta. Received: 6 November 1997 / Accepted:15 March 1998     

Effects of cytochalasin B on cytokinesis in mouse blastomeres. I. Light microscopic study

Mouse blastomeres exposed to cytochalasin B (CB) at subsequent stages of cytokinesis become spherical within 5–15 min. This change of form is preceded by rapid constriction of the furrow and by shape changes in the nonfurrow region such as blebbing, surface expansion, and formation of additional constrictions. While rounding up of blastomeres is completed within a few minutes in CB, 0.5 hr is the minimal duration of treatment that prevents resumption of cytokinesis after removal from CB.     

Manipulation of cytokinesis affects polar ionic current around the egg of Lymnaea stagnalis

Summary The fertilized egg of the mollusc Lymnaea stagnalis generates a polarized current pattern as measured with the vibrating probe. Here we investigated the basis of these polar ionic currents. Ionic currents were measured around eggs during the second meiotic division after interference with cytokinesis. Cytokinesis was either displaced by centrifugation or inhibited with cytochalasin or nocodazole. Furthermore, ectopic constrictions were induced with lectin treatment. It appeared that the inward current of the animal pole can be displaced by centrifugation and remains associated with the position of the meiotic apparatus. The influence of the meiotic apparatus on the polar current pattern seems to be directly related to membrane constrictions rather than to karyokinesis. This was demonstrated by a change in current density after induction of an ectopic constriction at the vegetal pole and by the abolishment of currents after cytochalasin treatment. Since the location of the outward current was not sensitive to centrifugation, it may be concluded that the vegetal outward current depends upon properties of the vegetal cortex. On the basis of these results, we conclude that the Lymnaea egg generates two types of ionic currents during the second meiotic division. The first is an inward current activated at the site of membrane constrictions. The second is an outward current associated with the vegetal cortex.     

Cleavage and membrane formation in the blastoderm of the dipteran Ceratitis capitata wied

In the Ceratitis capitata embryo, furrow formation follows the last mitosis divisions and leads to cellular blastoderm formation. This process displays several interesting features and requires the participation of bundles of microfilaments which are located at the furrow base at the onset of cytokinesis and contract synchronously, determining furrow growth. The new membranes for furrow growth seem to be largely provided by the fusion of many vesicles. Surface projections do not appear to contribute significantly to this phenomenon. At the end of cytokinesis the microfilaments are sandwiched between the plasma membrane and cisternae of endoplasmic reticulum. Subsequently the microfilaments disappear from the cytoplasmic side of the membrane but remain beneath the membranes of the connections and at the periphery of the yolk sack until gastrulation. On the basis of these observations some ultrastructural aspects of furrow formation and the role that the microfilaments may play during this process are discussed.     

Cell Constriction: Contractile Role of Microfilaments in Division and Development

The role of microfilaments in causing cell constrictions isdiscussed from a comparative point of view. Morphologicallysimilar microfilaments occur in the contractile ring of dividingcells and in the apices of neural plate cells during neurulation.New evidence is presented regarding the distribution and orientationof apical microfilaments in neural plates of chicks and salamanderembryos. These findings complement what is known about neurulationin frogs and cell cleavage in a variety of cells. In all cases,cell constriction occurs precisely and exclusively at thoseplanes in which circular arrays of microfilaments are found.A sliding mechanism of microfilament contractility is discussed,as are possible mechanisms involved in filament alignment. Attentionis given to the cell surface as a substratum for microfilamentassembly. New evidence is presented regarding the early morphologicaldetermination of the neural plate in Xenopus, even before microfilamentsare clearly evident or invagination begins.