An ultrastructural study of cytomixis in tobacco pollen mother cells

Intercellular chromatin migration (cytomixis) in the pollen mother cells of two tobacco (Nicotiana tabacum L.) lines was analyzed by electron microscopy during the first meiotic prophase. The maximal manifestation of cytomixis was observed in the pachytene. As a rule, several cells connected with one another by cytomictic channels wherein the nuclei migrated were observable at this stage. In the majority of cases, nuclei passed from cell to cell concurrently through several closely located cytomictic channels. Chromatin migrated between cells within the nuclear envelope, and its disintegration was unobservable. The nucleus, after passing through cytomictic channels into another cell, can be divided into individual micronuclei or, in the case of a direct contact with another nucleus, can form a nuclear bridge. It has been demonstrated that the chromatin structure after intracellular migration visually matches the chromatin structure before it passed through the cytomictic channel. No signs of pyknosis were observable in the chromatin of the micronuclei formed after cytomixis, and the synaptonemal complex was distinctly seen. The dynamics of changes in the nucleoli during cytomixis was for the first time monitored on an ultrastructural level. Possible mechanisms determining cytomixis are discussed and the significance of this process in plant development is considered.     

New insights into cytomixis: specific cellular features and prevalence in higher plants

The phenomenon of intercellular migration of nuclei in plant tissues (cytomixis) was discovered over a century ago, which has been followed by numerous attempts to clarify the essence of this process as well as to determine its causes and consequences. Most attention of researchers has been paid to cytomixis in microsporogenesis, since the transfer of part of genetic material between microsporocytes may influence the ploidy level of the produced pollen and, presumably, have an evolutionary significance. This review compiles the data on cytological pattern of cytomixis and proposes a scheme as to how cytomictic channels are formed and function in angiosperms. The prevalence of cytomixis in different plant taxa is analyzed using the published data. The causes, mechanisms, and consequences of the nuclear migration between cells in plant tissues are discussed.     

Cytochemical Localization of Adenosine Triphosphataes Activity During Cytomixis in Pollen Mother Cells of David Lily and Its Relation to the Intercellular Migrating Chromatin Substance

Standard lead precipitation procedures have been used to examine the localization of ATPase activity during cytomixis in pollen mother cells of Lilium davidii var. willmottiae (Wilson) Roffill. Before cytomixis, cells at this stage of development show ATPase activity on plasma membrane, in the endoplasmic reticulum, dictyosomes, plastids, plasmodesmata, and in part of the groundplasm; however, there is no ATPase activity on the chromatin and nucleolus. During cytomixis, the chromatin substance begin to transfer from one cell to an adjacent cell, reaction product indicating ATPase activity is observed associated with the chromatin and nucleolus. ATPase activity is also found with the cistenae of both endoplasmic reticulum and dictyosomes, and some plastids. There is no deposition of ATPase reaction product associated with the plasm membrane and intercellular spaces. After cytomixis, the chromatin is little or no deposition of enzyme reaction product. ATPase activity, however, is consistenlly found within the intercellular space and on the plasm membrane, and also occur in the endoplasmic reticulum, dictyosome and plastid. The presence or absence of ATPase activity in the cell structure of pollen mother cells before, during or after eytomixis is discussed in relation to the active uptake or export of water for short-distance transport. It is also suggested that the intensive ATPase activity in the nucleus during cytomixis of pollen mother cells is evidence for a transport system involved in the active movement of the intercellular migrating ebromatin substance.     

Migration of DNA-Containing Organelles between Tobacco Microsporocytes during Cytomixis

Ultrastructural analysis of intercellular migration of DNA-containing organelles (nuclei, mitochondria, and plastids) in tobacco microsporogenesis during cytomixis was conducted. It was demonstrated for the first time that the migrating part of the nucleus is covered with ribosomes and can contain the accumulation of nuclear pores. The possibility of mitochondrial migration between the plant cells was proven for the first time. It was demonstrated that mitochondria extremely rarely pass into neighboring cells, and their movement occurs through one cytomictic channel. In turn, plastids can generate the accumulations around cytomictic channels and actively migrate between the cells, even through small size cytomictic channels. It was established that plastids can pass into another cell through one or several cytomictic channels, and several plastids can also simultaneously migrate through one channel. The consequences of migration of DNA-containing organelles in the cells producing the pollen are discussed.     

The Relation between the Patterns of Cytomixis and Variation of Chromosome Numbers in Pollen Mother Cells of Jimsonweed ( Datura stramonium L.)

The phenomenon of cytomixis is a general occurenee in the pollen mother cells (PMCs) of certain angiosperms. Recently, we found this phenomenon also occurred in jimsonweed. According to our observations and a test of chi-square, it has been shown that the discrepancies between the patterns of intercellular chromatin migration and the variation of chromosome numbers in the PMCs of jimsonweed, as well as rye, are not significant. It means that the patterns of intercellulr chromatin migration and the variation chromosome numbers are related to each other.     

Cytomixis and its possible evolutionary role in a Kuwaiti population of Diplotaxis harra (Brassicaceae)

During a cytotaxonomical study of a Kuwaiti diploid (2 n  = 26) population of Diplotaxis harra (Forssk.) Boiss (Brassicaceae), cytomixis and aneuploidy were found in 1.5% and 7.8% of the pollen mother cells (PMCs), respectively. Cytomictic cells revealed wide variability as far as the number of involved PMCs and their stage of division were concerned. The cytoplasmic channels, which were of different sizes, contained migrating chromosomes in a few cases. In some PMCs, the location of some of the chromosomes was clearly indicative of an imminent migration from a donor to its attached recipient cell. At metaphase I and anaphase I, PMCs showed variable degree of chromosomal transfer. In addition to a genetic control of the phenomenon, stress factors such as high temperature or drought in certain periods of the growing season may have contributed to the incidence of cytomixis in the material. A possible relationship between cytomixis and detected aneuploid PMCs, as well as a reconsideration of an evolutionary role of cytomixis is discussed.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 143 , 169–175.     

百合花粉母细胞间细胞融合期间腺苷三磷酸酶活性的细胞化学定位及其与染色质胞间转移的关系

用标准的磷酸铅沉淀的细胞化学方法,对百合花粉母细胞间染色质穿壁运动期间及其前后三个时期中的腺苷三磷酸酶(ATP 酶)活性进行了超微结构的定位。结果表明:(1)在穿壁前,ATP 酶活性主要定位于质膜、胞间连丝及细胞间隙;在内质网、高尔基体、质体和某些局部的基质(groundplasm)中,也表现有 ATP 酶活性反应的产物;但在染色质和核仁中,一般都没有这种反应。(2)在穿壁时,染色质从一个细胞穿壁转移到另一个相邻细胞,同时看到染色质和核仁内出现密集的 ATP 酶活性反应产物;在内质网和高尔基体的腔内以及质体的片层上也产生明显的 ATP 酶活性反应;而在质膜、胞间连丝及细胞间隙内 ATP 酶活性明显降低,甚至看不到明显的活性反应。(3)在穿壁后,质膜及细胞间隙中又产生明显的 ATP 酶活性反应产物,但核内染色质上的 ATP 酶活性则显著降低,而核仁内则仍有较高的活性。同前二个时期一样,内质网、高尔基体和质体上的 ATP 酶仍表现明显的活性反应。最后讨论了三个不同发育时期 ATP 酶活性及其分布部位的改变与染色质胞间转移的关系。     

Induction of cytomixis affects microsporogenesis in Sesamum indicum L. (Pedaliaceae)

Cytomixis was recorded during microsporogenesis in sesame (Sesamum indicum L.), a member of the family Pedaliaceae. The phenomenon of cytomixis was observed at various stages of meiosis in 0.5% sodium azide (SA) treated populations of Sesamum indicum L. Cytomixis was observed to occur through various methods, i.e. by forming cytoplasmic channels and direct fusion of pollen mother cells (PMCs), the former was more frequent than the latter. The migration of nuclear content involved all the chromatin/chromosomes or part of it from donor to recipient cell/cells. Some completely empty meiocytes were also observed. Stickiness, precocious movement, laggards, unorientation and micronuclei were observed in almost all the sets treated with various doses of SA. Increase in the doses of SA had a positive effect on the percentage of PMCs showing cytomixis and chromosomal abnormalities. The impact of cytomixis on meiotic behaviour, reduced pollen viability and heterogeneous sized pollen grains were observed.     

Cytomixis in shoot apex of Norway spruce [<Emphasis Type="Italic">Picea abies</Emphasis> (L.) Karst.]

Atypical cell walls and nuclei were observed in the apex of Norway spruce shoots from late April to early May on the material collected from a few grafts of a clone of Norway spruce growing on an experimental area. Images of ultrastructure attest to cytomixis. The phenomenon of cytomixis has previously been described in various plant material, both in the meiotic and mitotic cells, but this is the first report of cytomixis in gymnosperms.     

Intra- and intertissular cytomictic interactions in the microsporogenesis of mono- and dicotyledonous plants

Comparative cytological analysis of intra- and intertissular cytomictic interactions in the microsporogenesis of mono- and dicotyledonous plants has been performed for two cellular systems: the microsporocytes and the tapetum. Cytomixis was shown to be more common for intratissular interactions, and cytomixis in the tapetum exhibited taxon-specific features, both structural and temporal. Nuclear migration in the microsporocytes mostly occurred during the zygotene–pachytene and exhibited certain synchrony with cytomixis in the tapetum. Intertissular cytomictic interactions (between the tapetum and the microsporocytes) were detected only in monocotyledonous plant anthers. Intertissular interactions may reflect more intense competition for space between the tapetum and the microsporocytes during the differentiation of anther tissues. The polyploid nuclei of the tapetum and the syncytia are powerful acceptors that can compete with the microsporocytes and attract the chromatin during translocation of the latter. The absence of intertissular interactions in dicotyledonous plants may be indicative of a better balance between the processes of differentiation of somatic and generative tissues of the microsporangium as compared to monocotyledonous plants.     

First detection of cytomixis and its consequences in <Emphasis Type="Italic">Thalictrum cultratum</Emphasis> Wall. (Ranunculaceae)

In current paper, we studied the meiotic chromosome number and details of cytomixis recorded for the first time in Thalictrum cultratum Wall. from the high altitude areas of northwest Himalaya (India). Cytomixis, the phenomenon of inter PMC (pollen mother cell) migration of chromatin material has been reported in many angiosperms and other groups of plants, but there is no published report in Thalictrum cultratum. All the presently studied accessions of the species existed at diploid level (x = 7). The present chromosome count of n = 14 has been reported for the first time from the study area. The cytological stability of any plant is an important consideration in view of its existence in nature. Meiotic disturbances can impose threat to the reproductive potential of plant. Current study reveals the occurrence of inter PMC migration of chromatin material. In present investigation, the phenomenon of cytomixis can be observed between 2 to 9 PMCs. Chromatin migration occurs through narrow and broad cytoplasmic channels or occasionally fused together during metaphase-I and anaphase-I. Cytomixis resulted in the formation of hypo- (n = 2, 2 + 1 small fragment, n = 3 + 3 small fragments, 4, 5 + extra mass of chromatin, 13) and hyperploid (n = 7 + 2 small fragments, 8 + 2 small fragments, 14, 14 + 2 small fragments, 15) PMCs, variable sized pollen grains and pollen sterility (20%). The current study reveals that cytomixis is responsible for formation of PMCs with variable chromosomes (hypoand hyperploid) and pollen grains of heterogeneous sizes and pollen sterility in T. cultratum. This is the first report of cytomixis in the species.     

高等植物中原生质细胞间运动研究的进展

20世纪初已在植物营养组织和生殖体中观察到细胞核或染色质在细胞间的穿壁迁移.但多年来涉及这一研究的学者大多倾向于将它看做是人为赝象而未予深究.50年代中期,自吴素萱在葱(Allium L.)、蒜(Allium sativum L.)等植物中重新发现了核穿壁运动现象以来,我国植物细胞与生理学家随之进行了系统与广泛的探讨.穿壁运动的研究及进展主要包括两个方面.一方面是营养组织中原生质的细胞间运动,由吴素萱、娄成后合作主持的有关研究项目着重与营养组织中有机物的运输、分配与再利用生理过程相联系.多年来积累的成果表明:1)核物质的穿壁运动是广泛存在于植物营养组织中的固有正常现象,是一定生理状态下有机物在细胞间运输与细胞内含物再分配的一种方式;2)通过对蒜芽鞘、小麦(Triticum aestivum L.)珠心、胚乳等组织的显微活体观察与亚显微结构分析,揭示与记录了原生质在细胞间运动的全过程与动态细节,发现参与穿壁过程的不限于核,核和胞质各组分可同时、亦可分别借自身的主动伸缩穿壁迁移;呈现穿壁运动的组织中,壁上部分胞间连丝结构剧烈变更,胞间通道明显扩大,可达100～300 nm,核或胞质可经此开放通道迁移而不致有损通道外沿质膜的完整;3)原生质穿壁运动的动力来自细胞的能量代谢,与微丝活动密切有关,穿壁的原生质组分呈现活跃的ATP酶活性.近期的研究表明,小麦胚乳细胞中作活跃伸缩运动的胞质纤索是由F肌动蛋白集束组成,肌动蛋白纤丝可跨胞分布而存在于常态胞间连丝中,从而使相邻细胞的胞质骨架得以实现胞间连续.另一方面是生殖体中染色质的穿壁运动研究,主要在郑国主持下开展,着重探讨花粉母细胞间染色质穿壁现象的普遍性及其与遗传、变异和进化的关系;对多种植物的系统观察证明,染色质穿壁出现在减数分裂前期Ⅰ的凝线期,染色质通过胞质通道在一系列细胞间依次迁移;由电镜观察可见,参与穿壁运动的还包括胞质的多种成员;穿壁运动的动力被认为由原生质的收缩蛋白提供.郑国等的研究成果表明,染色质穿壁运动可导致染色体数目、结构的改变,由此产生的部分有活力的配子体经受精后有可能导致新一代个体出现多倍体或非整倍体,引起遗传性状的变异.     

The role of chromatin structure in cell migration

Chromatin dynamics play a major role in regulating genetic processes. Now, accumulating data suggest that chromatin structure may also affect the mechanical properties of the nucleus and cell migration. Global chromatin organization appears to modulate the shape, the size and the stiffness of the nucleus. Directed-cell migration, which often requires nuclear reshaping to allow passage of cells through narrow openings, is dependent not only on changes in cytoskeletal elements but also on global chromatin condensation. Conceivably, during cell migration a physical link between the chromatin and the cytoskeleton facilitates coordinated structural changes in these two components. Thus, in addition to regulating genetic processes, we suggest that alterations in chromatin structure could facilitate cellular reorganizations necessary for efficient migration.     

Observations on the Rate of Early Embryogenesis and Cytomixis in Spring Wheat

The rate of early embryogenesis and cytomixis of spring wheat has been observed. The results obtained are summarized as follows: 2 h. after pollination, the two sperms entered into the egg nucleus and the polar nucleus respectively; after 6 h. triple fusion has been completed, a male nucleolus is discernible in egg nucleus. Twenty-four h. after pollination, 2-celled proembryo can be detectable, after 6 d the differentiation in some embryo has initiated. Three d after pollination, the formation of endosperm cells are proceeding, upto 6 d, the embryo sac are full of endosperm cells. After pollination through full development, the degeneration of the antipodals appears, 4 d later the structure of cell and nucleus disappeared. Only naked nucleoli and chromatin mass are remained. After 8 d the degeneration of antipodals almost has completed. Cytomixis has been seen in the cell of nucellus, endosperms, ovary, proembryo and differentiated embryo and it frequently appeared near the proembryo.     

Cytoplasmic Channels and Chromatin Migration in the Meiocytes of Arnebia hispidissima (Sieb.) DC

Microsporogenesis was studied in Arnebia hispidissima whichprovided interesting evidence for the occurrence of cytoplasmicchannels and DNA migration between the meiocytes. The intercommunicatingchannels (0.25–1.75 µ in diameter) are consideredto be a prerequisite for chromatin migration and to act as pathwaysbetween the connected meiocytes. They may develop at any stageof meiosis from early prophase to metaphase II and the migrationmay take place at any time following channel formation. Thetendency of chromatin to migrate seemed to be higher in theearly prophasic stages when it moved in the form of ill-definedmasses or clumps. The chances of migration as well as the amountof migrating chromatin is reduced as the meiosis advances. Theamount of chromatin migrating varies from a small portion tothe entire chromatin mass or nucleus. Pollen mother cells withdouble the normal amount of chromatin as well as those withlittle or none were studied as well as cells with increasedor decreased chromosome numbers. In the chromatin mass, uni-or bivalents stretched between the meiocytes or attenuated towardsthe channel, were frequently seen. Occasional semi-asyndesis,multivalent formation, precocity, and nondisjunction was alsonoted. The possibility of channel formation and the induction of chromatinmovement as a consequence of mechanical or procedural defectis ruled out. It is believed that these cytoplasmic channelshave no relation with the plasmodesmata but they arise de novoand that chromatin migration occurs as a natural phenomenon(in some species). The observations are not due to faulty fixation,staining, or squashing.     

Modeling of adhesion, protrusion, and contraction coordination for cell migration simulations

Cell migration is a highly complex, dynamical biological phenomenon that involves precise spatio-temporal coordination of distinctive sub-processes including adhesion, protrusion, and contraction of the cell. Observations of individual tumor cell migration reveal that cells generally exhibit either mesenchymal-type or amoeboid-type migration modes in native like environments. However, it has also been observed that some migrating cells are capable of morphologically adapting to their environment by modifying their type of migration. Recent studies suggest in fact that changes in biophysical and biomechanical properties of tumor cells can reversibly control their transition from one type of migration to the other. These changes may be caused by internal cell biomechanical mechanisms as well as mechanical and topological properties of the extracellular matrix. In order to understand the complex transition between the two modes and the role played by internal cellular mechanics during migration, we have developed a novel axisymmetric hyperviscoelastic cell model to simulate the dynamical behavior of a migrating cell. Numerical results from our study quantitatively demonstrate that the biomechanical properties of the cell may play an important role in the amoeboid-mesenchymal transition during migration. Our study will therefore not only help in creating a new platform for simulating cellular processes but will also provide insights into the role of sub-cellular mechanics in regulating various modes of migration during tumor invasion and metastasis.     

Calculation of the force field required for nucleus deformation during cell migration through constrictions

During cell migration in confinement, the nucleus has to deform for a cell to pass through small constrictions. Such nuclear deformations require significant forces. A direct experimental measure of the deformation force field is extremely challenging. However, experimental images of nuclear shape are relatively easy to obtain. Therefore, here we present a method to calculate predictions of the deformation force field based purely on analysis of experimental images of nuclei before and after deformation. Such an inverse calculation is technically non-trivial and relies on a mechanical model for the nucleus. Here we compare two simple continuum elastic models of a cell nucleus undergoing deformation. In the first, we treat the nucleus as a homogeneous elastic solid and, in the second, as an elastic shell. For each of these models we calculate the force field required to produce the deformation given by experimental images of nuclei in dendritic cells migrating in microchannels with constrictions of controlled dimensions. These microfabricated channels provide a simplified confined environment mimicking that experienced by cells in tissues. Our calculations predict the forces felt by a deforming nucleus as a migrating cell encounters a constriction. Since a direct experimental measure of the deformation force field is very challenging and has not yet been achieved, our numerical approaches can make important predictions motivating further experiments, even though all the parameters are not yet available. We demonstrate the power of our method by showing how it predicts lateral forces corresponding to actin polymerisation around the nucleus, providing evidence for actin generated forces squeezing the sides of the nucleus as it enters a constriction. In addition, the algorithm we have developed could be adapted to analyse experimental images of deformation in other situations.     

The granulocyte nucleus and lamin B receptor: avoiding the ovoid

The major human blood granulocyte, the neutrophil, is an essential component of the innate immunity system, emigrating from blood vessels and migrating through tight tissue spaces to the site of bacterial or fungal infection where they kill and phagocytose invading microbes. Since the late nineteenth century, it has been recognized that the human neutrophil nucleus is distinctly not ovoid as in other cell types, but possesses a lobulated (segmented) shape. This deformable nucleus enhances rapid migration. Recent studies have demonstrated that lamin B receptor (LBR) is necessary for the non-ovoid shape. LBR is an integral membrane protein of the nuclear envelope. A single dominant mutation in humans leads to neutrophils with hypolobulated nuclei (Pelger–Huet anomaly); homozygosity leads to ovoid granulocyte nuclei. Interestingly, LBR is also an enzyme involved in cholesterol metabolism. Homozygosity for null mutations is frequently lethal and associated with severe skeletal deformities. In addition to the necessity for LBR, formation of the mature granulocyte nucleus also depends upon lamin composition and microtubule integrity. These observations are part of a larger question on the relationships between nuclear shape and cellular function.     

Induced cytomictic variations and syncyte formation during microsporogenesis in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.

The intercellular translocation of chromatin material along with other cytoplasmic contents among the proximate meiocytes lying in close contact with each other commonly referred as cytomixis was reported during microsporogenesis in Phaseolus vulgaris L., a member of the family Fabaceae. The phenomenon of cytomixis was observed at three administered doses of gamma rays viz. 100, 200, and 300 Gy respectively in the diploid plants of Phaseolus vulgaris L. The gamma rays irradiated plants showed the characteristic feature of inter-meiocyte chromatin/chromosomes transmigration through various means such as channel formation, beak formation or by direct adhesion between the PMC’s (Pollen mother cells). The present study also reports the first instance of syncyte formation induced via cytomictic transmigration in Phaseolus vulgaris L. Though the frequency of syncyte formation was rather low yet these could play a significant role in plant evolution. It is speculated that syncyte enhances the ploidy level of plants by forming 2n gametes and may lead to the production of polyploid plants. The phenomenon of cytomixis shows a gradual inclination along with the increasing treatment doses of gamma rays. The preponderance of cytomixis was more frequent during meiosis I as compared to meiosis II. An interesting feature noticed during the present study was the channel formation among the microspores and fusion among the tetrads due to cell wall dissolution. The impact of this phenomenon is also visible on the development of post-meiotic products. The formation of heterosized pollen grains; a deviation from the normal pollen grains has also been reported. The production of gametes with unbalanced chromosomes is of utmost importance and should be given more attention in future studies as they possess the capability of inducing variations at the genomic level and can be further utilized in the improvement of germplasm.