Nuclear Cytoplasmic Domains Research Articles

Effect of estrone on somatic and female gametophyte cell division and differentiation in Arabidospis thaliana cultured in vitro

The aim of the study was to determine the effect of the mammalian female sex hormone estrone on differentiation of somatic tissues and on induction of autonomous endosperm in culture of female gametophyte cells of Arabidopsis thaliana ecotype Columbia (Col-0). In culture, estrone-stimulated development of autonomous endosperm (AE) occurred in 14.7% of unpollinated pistils. The AE represented development stages similar to those of young endosperm after fertilization and AE of fis mutants in vivo. In the majority of ovules the AE was in a few-nucleate young stage. Some ovules showed more advanced stages of AE development, with nuclei and cytoplasm forming characteristic nuclear cytoplasmic domains (NCDs). Sporadically, AE was divided into regions characteristic for Arabidopsis endosperm formed after fertilization. Direct organogenesis (caulogenesis, rhizogenesis), callus proliferation and formation of trichome-like structures were observed during in vitro culture of hypocotyls and cotyledons of 3-day-old seedlings cultured on medium supplemented with estrone for 28 days. Histological analysis showed adventitious root formation and changes in explant anatomy caused by estrone.

Occurrence and cytological mechanism of numerically unreduced pollen in diploid Populus euphratica

AbstractUnreduced gametes are the driving force for the polyploidizaiton of plants in nature, and are also an important tool for breeding of triploid individuals. The final heterozygosity of a 2n pollen grain depends on the cytological mechanism behind 2n pollen formation. In this study, meiotic abnormalities were analysed using fluorescent chromosome staining and indirect immunofluorescence during the microsporogenesis of 18 genotypes of diploid P. euphratica Oliv. (2n = 2x = 38). Among the 18 genotypes, 16 genotypes produce 2n pollen and two genotypes produce only normal n pollen. In all 2n pollen producers, we found that the first meiotic division was normal but that the second division was characterized by frequent abnormal spindle orientation (parallel, tripolar, and fused spindles) and premature cytokinesis. The parallel, fused spindles and premature cytokinesis were considered to be leading dyad formation, and tripolar spindles seemed to be causing triad formation at the tetrad stage. There was a higher frequency of parallel spindles than other spindle forms, but no significant correlations between parallel spindles and dyads were observed. However, a significant association (r = 0.68, P < 0.05) between the tripolar spindles and dyads was found. In some Microspore mother cells (MMCs), an indirect immunofluorescence examination of meiosis II revealed that the parallel spindles led to the gathering of one or two non-sister groups of chromosomes, causing an incorporation of RMSs from two daughter nuclei. Therefore, the incorporated RMSs established two nuclear cytoplasmic domains for the control of division plane, resulting in either triad or dyad formation.

Cytological characteristics of numerically unreduced pollen production in Populus tomentosa Carr.

Natural triploids of Populus tomentosa (2n = 3x = 57) are presumably the result of sexual polyploidization through the union of normal n female gametes and numerically unreduced (2n) male gametes. In our microscopic study of microspore mother cells (MMCs) of diploid P. tomentosa (2n = 2x = 38), we observed that the first meiotic division was normal but that the second division was characterized by frequent abnormal spindle orientation (parallel, tripolar, and fused spindles) and premature cytokinesis. The parallel, fused spindles and premature cytokinesis were considered to be leading dyad formation, and tripolar spindles seemed to be leading triad formation at the tetrad stage. There was a higher frequency of parallel spindles than other spindle forms, but there were no significant correlations between parallel spindles and dyads. An indirect immunofluorescence examination of meiosis II revealed that four tetragonally arranged nuclei were formed in MMCs with parallel spindles and that there were radial microtubules systems (RMSs) among these four nuclei, leading to the tetragonal tetrad. In some MMCs, however, the parallel spindles led to the gathering of one or two non-sister groups of chromosomes, causing an incorporation of RMSs from two daughter nuclei. Thus, the incorporated RMSs established three or two nuclear cytoplasmic domains for the control of division plane, resulting in either triad or dyad formation. These results provide new insights on the mechanism of parallel spindles leading to numerically unreduced pollen formation and on the selection and utilization of this type of pollen in polyploid breeding of P. tomentosa.

Arabidopsis Separase Functions beyond the Removal of Sister Chromatid Cohesion during Meiosis

Separase is a capase family protease that is required for the release of sister chromatid cohesion during meiosis and mitosis. Proteolytic cleavage of the alpha-kleisin subunit of the cohesin complex at the metaphase-to-anaphase transition is essential for the proper segregation of chromosomes. In addition to its highly conserved role in cleaving the alpha-kleisin subunit, separase appears to have acquired additional diverse activities in some organisms, including involvement in mitotic and meiotic anaphase spindle assembly and elongation, interphase spindle pole body positioning, and epithelial cell reorganization. Results from the characterization of Arabidopsis (Arabidopsis thaliana) separase (ESP) demonstrated that meiotic expression of ESP RNA interference blocked the proper removal of cohesin from chromosomes and resulted in the presence of a mixture of fragmented chromosomes and intact bivalents. The presence of large numbers of intact bivalents raised the possibility that separase may also have multiple roles in Arabidopsis. In this report, we show that meiotic expression of ESP RNA interference blocks the removal of cohesin during both meiosis I and II, results in alterations in nonhomologous centromere association, disrupts the radial microtubule system after telophase II, and affects the proper establishment of nuclear cytoplasmic domains, resulting in the formation of multinucleate microspores.

Microtubules in early development of the megagametophyte of Ginkgo biloba

Food storage tissue in the seeds of gymnosperms is female gametophyte (megagametophyte) that develops before fertilization, whereas, in seeds of angiosperms, food is stored as endosperm initiated by double fertilization. The megagametophyte is haploid, and endosperm is usually triploid, at least initially. Despite differences in origin, ploidy level, and developmental trigger, the early events of female gametophyte development in ginkgo are very similar to nuclear endosperm development in the seeds of angiosperms. In both, development begins as a single cell that undergoes multiple mitoses without cytokinesis, to produce a large syncytium. This study provided evidence that microtubule involvement in organization of the syncytium into nuclear cytoplasmic domains (NCDs) via nuclear-based radial microtubule systems is a critical developmental feature in the ginkgo megagametophyte, as it is in endosperm. Once the initial anticlinal walls have been deposited at the boundaries of NCDs, cellularization proceeds by the process of alveolation. Continued unidirectional growth of the alveolar walls is an outstanding example of polar cytokinesis. Ginkgo megagametophyte development appears to occur uniformly throughout the entire chamber, whereas nuclear type endosperm usually exhibits distinct developmental domains. These observations suggest that there is a fundamental pathway for the development and cellularization of syncytia in seed development.

The Pleiomorphic Plant MTOC: An Evolutionary Perspective

Abstractγ‐Tubulin is an essential component of the microtubule organizing center (MTOC) responsible for nucleating microtubules in both plants and animals. Whereas γ‐tubulin is tightly associated with centrosomes that are inheritable organelles in cells of animals and most algae, it appears at different times and places to organize the myriad specialized microtubule systems that characterize plant cells. We have traced the distribution of γ‐tubulin through the cell cycle in representative land plants (embryophytes) and herein present data that have led to a concept of the pleiomorphic and migratory MTOC. The many forms of the plant MTOC at spindle organization constitute pleiomorphism, and stage‐specific “migration” is suggested by the consistent pattern of redistribution of γ‐tubulin during mitosis. Mitotic spindles may be organized at centriolar centrosomes (only in final divisions of spermatogenesis), polar organizers (POs), plastid MTOCs, or nuclear envelope MTOCs (NE‐MTOCs). In all cases, with the possible exception of centrosomes in spermatogenesis, the γ‐tubulin migrates to broad polar regions and along the spindle fibers, even when it is initially a discrete polar entity. At anaphase it moves poleward, and subsequently migrates from polar regions (distal nuclear surfaces) into the interzone (proximal nuclear surfaces) where interzonal microtubule arrays and phragmoplasts are organized. Following cytokinesis, γ‐tubulin becomes associated with nuclear envelopes and organizes radial microtubule systems (RMSs). These may exist only briefly, before establishment of hoop‐like cortical arrays in vegetative tissues, or they may be characteristic of interphase in syncytial systems where they serve to organize the common cytoplasm into nuclear cytoplasmic domains (NCDs).

Temporal and spatial appearance of wall polysaccharides during cellularization of barley (Hordeum vulgare) endosperm

Barley endosperm begins development as a syncytium where numerous nuclei line the perimeter of a large vacuolated central cell. Between 3 and 6 days after pollination (DAP) the multinucleate syncytium is cellularized by the centripetal synthesis of cell walls at the interfaces of nuclear cytoplasmic domains between individual nuclei. Here we report the temporal and spatial appearance of key polysaccharides in the cell walls of early developing endosperm of barley, prior to aleurone differentiation. Flowering spikes of barley plants grown under controlled glasshouse conditions were hand-pollinated and the developing grains collected from 3 to 8 DAP. Barley endosperm development was followed at the light and electron microscope levels with monoclonal antibodies specific for (1-->3)-beta-D: -glucan (callose), (1-->3,1-->4)-beta-D: -glucan, hetero-(1-->4)-beta-D: -mannans, arabino-(1-->4)-beta-D: -xylans, arabinogalactan-proteins (AGPs) and with the enzyme, cellobiohydrolase II, to detect (1-->4)-beta-D: -glucan (cellulose). Callose and cellulose were present in the first formed cell walls between 3 and 4 DAP. However, the presence of callose in the endosperm walls was transient and at 6 DAP was only detected in collars surrounding plasmodesmata. (1-->3,1-->4)-beta-D: -Glucan was not deposited in the developing cell walls until approximately 5 DAP and hetero-(1-->4)-beta-D: -mannans followed at 6 DAP. Deposition of AGPs and arabinoxylan in the wall began at 7 and 8 DAP, respectively. For arabinoxylans, there is a possibility that they are deposited earlier in a highly substituted form that is inaccessible to the antibody. Arabinoxylan and heteromannan were also detected in Golgi and associated vesicles in the cytoplasm. In contrast, (1-->3,1-->4)-beta-D: -glucan was not detected in the cytoplasm in endosperm cells; similar results were obtained for coleoptile and suspension cultured cells.

γ-Tubulin and microtubule organization during microsporogenesis in Ginkgo biloba

This is the first report on gamma-tubulin and microtubule arrays during microsporogenesis in a gymnosperm. Meiosis in Ginkgo biloba is polyplastidic, as is typical of the spermatophyte clade, and microtubule arrays are organized at various sites during meiosis and cytokinesis. In early prophase, a cluster of gamma-tubulin globules occurs in the central cytoplasm adjacent to the off-center nucleus. These globules diminish in size and spread over the surface of the nucleus. A system of microtubules focused on the gamma-tubulin forms a reticulate pattern in the cytoplasm. As the nucleus migrates to the center of the microsporocyte, gamma-tubulin becomes concentrated at several sites adjacent to the nuclear envelope. Microtubules organized at these foci of gamma-tubulin give rise to a multipolar prophase spindle. By metaphase I, the spindle has matured into a distinctly bipolar structure with pointed poles. In both first and second meiosis, gamma-tubulin becomes distributed throughout the metaphase spindles, but becomes distinctly polar again in anaphase. In telophase I, gamma-tubulin moves from polar regions to the proximal surface of chromosome groups/nuclei where interzonal microtubules are organized. No cell wall is deposited and the interzonal microtubules embrace a plate of organelles between the two nuclear cytoplasmic domains (NCDs) of the dyad. Following second meiosis, phragmoplasts that form between sister and non-sister nuclei fuse to form a complex six-sided structure that directs simultaneous cytokinesis. Gamma-tubulin becomes associated with nuclei after both meiotic divisions and is especially conspicuous in the distal hemisphere of each young microspore where an unusual encircling system of cortical microtubules develops.

Events during the first four rounds of mitosis establish three developmental domains in the syncytial endosperm of Arabidopsis thaliana.

Endosperm begins development as a single fertilized cell that undergoes many rounds of mitosis without cytokinesis resulting in a syncytium. The multinucleate cytoplasm is organized by nucleus-based radial microtubule systems into nuclear-cytoplasmic domains. When microtubules are organized into mitotic spindles, the integrity of the common cytoplasm is maintained by an unaltered network of filamentous actin. The first four rounds of mitosis result in the establishment of three developmental domains within the common cytoplasm. The spindles of the first two rounds of mitosis are oriented parallel to the long axis of the central cell, resulting in four nuclear-cytoplasmic domains in a filamentous arrangement. A switch in spindle orientation occurs in the third round of mitosis; all four spindles are oriented perpendicular to the long axis resulting in eight nuclear-cytoplasmic domains arranged in two adjacent files. Whereas the first three rounds of mitosis are synchronous, the fourth occurs as a wave of successive mitoses that begins at the micropylar pole. By the 16-nuclei stage, differences in nuclear shape, cytoskeletal arrays, and cytoplasmic characteristics mark the differentiation of the syncytium into micropylar, central, and chalazal developmental chambers. Nuclei in the micropylar chamber are fusiform and sheathed by parallel microtubules that flare from their tips, while those in the central and chalazal chambers are spherical. Nuclei in the central chamber are surrounded by radial microtubule systems, while those in the chalaza are enmeshed in a reticulum of microtubules. Whereas the cytoplasm in both micropylar and chalazal chambers is dense and nearly nonvacuolate, the syncytium in the central chamber consists of a single layer of evenly spaced nuclear-cytoplasmic domains surrounding a large central vacuole.

Cytoskeletal organization of the micropylar endosperm in Coronopus didymus L. (Brassicaceae).

The micropylar chamber of the mustard Coronopus didymus is a developmental domain distinct from the contiguous central chamber and the more extreme chalazal chamber. Early in syncytial development the micropylar endosperm surrounding the embryo becomes populated with unusual fusiform to multilobed nuclei. These nuclei are sheathed by unique parallel arrays of microtubules that focus at tips of the nuclei and flare to connect with a reticulate network in the common cytoplasm. F-actin does not closely invest the nuclei but instead forms an extensive but separate cytoplasmic reticulum. When the embryo is in the early heart stage, the cytoskeleton of the endosperm undergoes a remarkable transition in preparation for cellularization. Microtubules become reorganized into radial arrays emanating from the nuclei, which themselves become spherical. Radial microtubule systems (RMSs), which replace both the parallel microtubules and the cytoplasmic reticulum, organize the common cytoplasm into evenly spaced nuclear cytoplasmic domains (NCDs). F-actin gradually becomes coaligned with the RMSs. Phragmoplasts are initiated adventitiously at the interfaces of opposing RMSs in the absence of mitosis. Cell plate deposition, which is initiated at multiple sites, results in a network of walls formed more or less simultaneously around the densely packed NCDs. The walls, which are rich in 1-3-beta-glucans, join with one another and with the existing walls of both the central cell and embryo to complete cellularization in the micropylar chamber. In the adjacent central chamber where the syncytium is restricted to a thin peripheral layer by the large central vacuole, basic organization of the syncytium into NCDs is followed by alternating cycles of alveolation and periclinal cell division resulting in cellularization.

Phragmoplasts in the Absence of Nuclear Division

All land plants (embryophytes) use a phragmoplast for cytokinesis. Phragmoplasts are distinctive cytoskeletal structures that are instrumental in the deposition of new walls in both vegetative and reproductive phases of the life cycle. In meristems, the phragmoplast is initiated among remaining non-kinetochore spindle fibers between sister nuclei and expands to join parental walls at the site previously marked by the preprophase band of microtubules (PPB). The microtubule cycle and cell cycle are closely coordinated: the hoop-like cortical microtubules of interphase are replaced by the PPB just prior to prophase, the PPB disappears as the spindle forms, and the phragmoplast mediates cell plate deposition after nuclear division. In the reproductive phase, however, cortical microtubules and PPBs are absent and cytokinesis may be uncoupled from the cell cycle resulting in multinucleate cells (syncytia). Minisyncytia of 4 nuclei occur in microsporocytes and several (typically 8) nuclei occur in the developing megagametophyte. Macrosyncytia with thousands of nuclei may occur in the nuclear type endosperm. Cellularization of syncytia involves formation of adventitious phragmoplasts at boundaries of nuclear-cytoplasmic domains (NCDs) defined by radial microtubule systems (RMSs) emanating from non-sister nuclei. Once initiated in the region of microtubule overlap at interfaces of opposing RMSs, the adventitious phragmoplasts appear structurally identical to interzonal phragmoplasts. Phragmoplasts are constructed of multiple opposing arrays similar to what have been termed microtubule converging centers. The individual phragmoplast units are distinctive fusiform bundles of anti-parallel microtubules bisected by a dark mid-zone where vesicles accumulate and fuse into a cell plate.

Patterns of Cytoskeletal Organization Reflect Distinct Developmental Domains in Endosperm ofCoronopus didymus(Brassicaceae)

New data on the interrelationships of F‐actin and microtubules during endosperm development reveal distinct domains in the micropylar chamber (MC) containing the embryo, the large curved central chamber (CC), and the small chalazal chamber (ChC). Reported for the first time in any endosperm are the following: (1) data on the interrelationships between microtubules and F‐actin during development in all three chambers, and (2) an early stage of syncytial endosperm characterized by unusual fusiform to multangular nuclei sheathed by parallel arrays of microtubules. As is characteristic of nuclear endosperm development, the common cytoplasm is organized into nuclear cytoplasmic domains (NCDs) defined by radial microtubule systems, which determine placement of walls at the cellularization stage. Both microtubules and F‐actin are reorganized in preparation of the syncytium for simultaneous cytokinesis. Cellularization directly follows formation of adventitious phragmoplasts in the MC but is delayed in the CC as ...

Development of endosperm in Arabidopsis thaliana

The process of endosperm development in Arabidopsis was studied using immunohistochemistry of tubulin/microtubules coupled with light and confocal laser scanning microscopy. Arabidopsis undergoes the nuclear type of development in which the primary endosperm nucleus resulting from double fertilization divides repeatedly without cytokinesis resulting in a syncytium lining the central cell. Development occurs as waves originating in the micropylar chamber and moving through the central chamber toward the chalazal tip. Prior to cellularization, the syncytium is organized into nuclear cytoplasmic domains (NCDs) defined by nuclear-based radial systems of microtubules. The NCDs become polarized in axes perpendicular to the central cell wall, and anticlinal walls deposited among adjacent NCDs compartmentalize the syncytium into open-ended alveoli overtopped by a crown of syncytial cytoplasm. Continued centripetal growth of the anticlinal walls is guided by adventitious phragmoplasts that form at interfaces of microtubules emanating from adjacent interphase nuclei. Polarity of the elongating alveoli is reflected in a subsequent wave of periclinal divisions that cuts off a peripheral layer of cells and displaces the alveoli centripetally into the central vacuole. This pattern of development via alveolation appears to be highly conserved; it is characteristic of nuclear endosperm development in angiosperms and is similar to ancient patterns of gametophyte development in gymnosperms.

Development of the endosperm in rice (Oryza sativa L.): Cellularization

The syncytial endosperm of rice undergoes cellularization according to a regular morphogenetic plan. At 3 days after pollination (dap) mitosis in the peripheral synctium ceases. Radial systems of microtubules emanating from interphase nuclei define nuclear-cytoplasmic domains (NCDs) which develop axes perpendicular, to the embryo sac wall. Free-growing anticlinal walls between adjacent NCDs compart-mentalize the cytoplasm into open-ended alveoli which are overtopped by syncytial cytoplasm adjacent to the central vacuole. At 4 dap, mitosis resumes as a wave originating adjacent to the vascular bundle. The spindles are oriented parallel to the alveolar walls and cell plates formed in association with interzonal phragmoplasts result in periclinal walls that cut off a peripheral layer of cells and an inner layer of alveoli displaced toward the center. Polarized growth of the newly formed alveoli and elongation of the anticlinal walls occurs during interphase. The next wave of cell division in the alveoli proceeds as the first and a second cylinder of cells is cut off inside the peripheral layer. The periods of polarized growth/anticlinal wall elongation alternating with periclinal cell division are repeated 3–4 times until the grain is filled by 5 dap.

Polarization predicts the pattern of cellularization in cereal endosperm

The endosperm of cereal grains develops as a multinucleate mass of wall-less cytoplasm (syncytium) that lines the periphery of the central cell before becoming cellular. The pattern of initial wall formation is precisely oriented and is followed by a round of precisely oriented formative cell division that gives rise to initials for the two tissues of endosperm. The initial anticlinal walls form at boundaries of nuclear-cytoplasmic domains (NCDs) defined by radial microtubules emanating from nuclei in the syncytium. Polarized growth of the NCDs in axes perpendicular to the embryo sac wall and centripetal elongation of the anticlinal walls results in a single layer of open ended alveoli overtopped by the remaining syncytial cytoplasm. This arboreal stage, so named because the elongate nucleate columns of cytoplasm resemble an orchard of trees, predicts the division polarity of the imminent formative division. Mitosis occurs as a wave which, like polarization, moves in both directions from ventral to dorsal. Spindles are oriented parallel to the long axis of the alveoli and cell plates give rise to periclinal walls. The outer daughter nuclei (aleurone initials) are thus completely enclosed by walls and the inner nuclei (starchy endosperm initials) are in alveoli adjacent to the central vacuole.

Nuclear cytoplasmic domains, microtubules and organelles in microsporocytes of the slipper orchid Cypripedium californicum A. Gray dividing by simultaneous cytokinesis

Nuclear cytoplasmic domains, microtubules and organelles in microsporocytes of the slipper orchid Cypripedium californicum A. Gray dividing by simultaneous cytokinesis

Nuclear cytoplasmic domains, microtubules and organelles in microsporocytes of the slipper orchid Cypripedium californicum A. Gray dividing by simultaneous cytokinesis

Microsporocytes of the slipper orchidCypripedium californicum A. Gray divide simultaneously after second meiosis. The organization and apportionment of the cytoplasm throughout meiosis are functions of nuclear-based radial microtubule systems (RMSs) that define domains of cytoplasm - a single sporocyte domain before meiosis, dyad domains within the undivided cytoplasm after first meiosis, and four spore domains after second meiosis. Organelles migrate to the interface of dyad domains in the undivided cytoplasm after first meiotic division, and second meiotic division takes place simultaneously on both sides of the equatorial organelle band. Microtubules emanating from the telophase II nuclei interact to form columnar arrrays that interconnect all four nuclei, non-sister as well as sister. Cell plates are initiated in these columns of microtubules and expand centrifugally along the interface of opposing RMSs, coalescing in the center of the sporocyte and joining with the original sporocyte wall at the periphery to form the tetrad of microspores. Organelles are distributed into the spore domains in conjunction with RMSs. These data, demonstrating that cytokinesis in microsporogenesis can occur in the absence of both components of the typical cytokinetic apparatus (the preprophase band of microtubules which predicts the division site and the phragmoplast which controls cell-plate deposition), suggest that plant nuclei have an inherent ability to establish a domain of cytoplasm via radial microtubule systems and to regulate wall deposition independently of the more complex cytokinetic apparatus of vegetative cells.

Endosperm Development in Barley: Microtubule Involvement in the Morphogenetic Pathway.

An immunofluorescence study of sectioned barley endosperm imaged by confocal laser scanning microscopy provided three-dimensional data on the relationship of microtubules to the cytoplasm, nuclei, and cell walls during development from 4 to 21 days after pollination (DAP). Microtubules play an important role throughout endosperm ontogeny. The syncytium is organized into units of nuclear-cytoplasmic domains by nuclear-based radial microtubule systems that appear to control the pattern of the first anticlinal walls at 5 to 6 DAP. After 7 DAP, phragmoplasts of two origins (interzonal and cytoplasmic) guide wall formation. Large compartments formed by the "free growing" walls in association with cytoplasmic phragmoplasts formed adventitiously at interfaces of opposing microtubule systems are subsequently subdivided by interzonal phragmoplast/cell plates to give rise to the starchy endosperm. During development of the aleurone layer from 8 to 21 DAP, the microtubule cycle is typical of plant histogenesis; cortical microtubules are hooplike, and preprophase bands of microtubules predict the division plane.

The role of microtubules in establishing nuclear spatial patterns in multinucleate green algae

In uninucleate cells, cytokinesis follows karyokinesis, thereby reestablishing a specific nucleus-to-cytoplasm ratio. In multinucleate cells, cytokinesis is absent or infrequent; no plasmalemma boundary defines the cytoplasmic territory of an individual nucleus. Several genera of large multinucleate green algae were examined with epifluorescence light microscopy to determine whether the patterns of cytoplasmic organization establish nuclear cytoplasmic domains. Randomly spaced nuclei, singular mitotic events and cytoplasmic streaming characterize the organization of two genera,Derbesia andBryopsis (Caulerpales). The cells ofValonia, Valoniopsis, Boergesenia, Ventricaria (Siphonocladales), andHydrodictyon (Chlorococcales) display regularly spaced nuclei which undergo synchronous divisions in a stationary cytoplasm. In the cytoplasm of genera with regularly spaced nuclei, microtubules radiate from all nuclei in late telophase-early interphase. These internuclear microtubule arrays are not found in algal genera with randomly spaced nuclei. It is hypothesized that these microtubule arrays play a role in establishing the cytoplasmic domain of each nucleus in genera with regularly spaced nuclei. Loss of microtubule arrays during the events of mitosis correlated positively with the increasing randomization of nuclear patterns in algae grown in microtubule inhibitors. Cytoplasmic domains were maintained when cells were grown in the same media in the dark. This suggests that, after a round of division, regular nuclear spacing in certain multinucleate algae is reestablished by internuclear microtubule arrays, which are not, however, required to maintain spacing during interphase.