Cytoplasmic differentiation during tetrad formation and early microspore development inImpatiens sultani

Summary From early prophase stage until probaculae formation within the tetrad stage considerable cytoplasmic changes occur. The changes merely concern the ribosome population, the plasma matrix and, the endomembrane system formed by endoplasmic reticulum, dictyosomes and dictyosome-vesicles.The ultrastructure and morphology of mitochondria and plastids remain fairly unchanged, apart from the mobilization of starch during primexine formation.During meiotic prophase there is an increase in ribosome number, accompanied by the presence of nucleoloids in the cytoplasm. Simultaneously the electron density of the cytoplasm strongly increases, indicating a fair increase in protein content. Nucleoloids are also observed in the cytoplasm after primexine formation, accompanied by localized accumulation of ribosomes. Up to the individualization of the microspores the dictyosomes are in an inactive state. After that, they become very active, especially during primexine formation when numerous large dictyosome-vesicles are present.The endoplasmic reticulum (ER), initially in a plate-like configuration, disappears from the cytoplasm during primexine formation. Abundant, smooth and tubular ER is present when probaculum formation starts.     

Pre-fertilization ovule development inCapsella: ultrastructure and ultracytochemical localization of acid phosphatase in the meiocyte

Summary Pre-meiotic and prophase I ovules ofCapsella bursa-pastoris (L.) Medic.(monosporic,Polygonum type of gametophyte development) were fixed routinely or incubated in a modified Gomori medium containing -glycerophosphate as a substrate. Prior to the beginning of meiosis the potential meiocyte is ultrastructurally similar to the other cells of the nucellus and is distinguished only by its size and position. At the initiation of prophase I dramatic ultrastructural and ultracytochemical changes take place in the female meiocyte. These include the sudden appearance of cytoplasmic structures composed of single and multiple concentric cisternae, distinctive changes in plastids and mitochondria, and the blebbing of 0.3 m double-membraned vesicles from the nuclear envelope. The concentric cisternae encapsulate portions of cytoplasm containing ribosomes, plastids, mitochondria, ER fragments and vesicles. Both single and multiple concentric cisternae localize high levels of acid phosphatase and function as autophagic vesicles (AVs) that sequester ribosomes and organelles for destruction during meiosis. Plastids stop dividing and become more spherical during prophase I. Some plastids localize acid phosphatase and many show continuities between the outer membrane and the plastid envelope and acid phosphatase-rich RER cisternae. Mitochondria appear as dense, contracted spheres or rods. Some mitochondria localize acid phosphatase but they do not show membrane confluencies with the ER. Some of the plastids and mitochondria that are segregated into the functional megaspore at meiosis II are destroyed but others apparantly survive meiosis and give rise to the plastid and mitochondrial populations of the young gametophyte (Schulz andJensen, unpublished). The lateral and end walls of the meiocyte show patches of intense aniline blue fluorescence and the chalazal end wall of the cell is perforated with large numbers of plasmodesmata.Research supported by NSF Grant PCM-79-11018. The authors gratefully acknowledge the valuable assistance of David Lee Ivans in this project.     

Ribosomes, membranes and organelles during meiosis in angiosperms.

Evidence is presented from both observational and analytical techniques indicating profound changes to take place in the ribosome population during male and female meiosis in some flowering plants. During microsporogenesis these appear to involve the elimination of the major part of the ribosome complement early in the meiotic prophase, and its subsequent restoration by the disintegration in the tetrad cytoplasm of 'nucleoloids', themselves synthesized in the nucleus during late prophase. In female tissue the process is essentially similar except for differences in the restoration of the ribosome population. Immediately before the eradication of the ribosomes in both sexes, a sizeable proportion of the meiocyte cytoplasm is encapsulated by double, or multiple unit membrane profiles. Significantly this cytoplasm remains unaffected by the agents responsible for the degredation of the ribosome population. These events are also reflected in the organelle populations where cycles of dedifferentiation and redifferentiation take place. In view of evidence from other organisms, it is considered unlikely that these cycles are in any way a prerequisite of meiosis, but more a characteristic of cells undergoing major changes of phase, where a cytoplasmic 'clean-up' is required before the next stage of growth may begin.     

Ultrastructural Aspects of Sporogenesis in a Fern, Pteridium aquilinum (L.) Kuhn

The meiotic process of sporogenesis in Pteridium is accompaniedby a regular cycle of ultrastructural events. These includethe dedifferentiation and redifferentiation of organelles, andthe elimination and restoration of ribosomes. Although similarin nature to corresponding stages in other land plants, sporogenesisin Pteridium provides several unique features. During prophase the plastids pass through a stage during whichno envelope can be resolved at the periphery of the stroma.They appear to survive this period and there is no evidencethat plastid lineage is broken from primary archesporium tospore. The plastid cycle is accompanied by the presence of theiron-protein complex phytoferritin. The repopulation of the spore cytoplasm with ribosomes, followingthe prophase elimination, is achieved through the agency ofnucleoloid-like bodies. These bodies, however, are not synthesizedwithin the nucleus, but form within membrane-bound regions ofthe early prophase cytoplasm. Prophase is also characterized by the development of nuclearvacuoles, expansions of the perinuclear space, which progressivelyramify through the karyoplasm prior to the dissolution of thenuclear envelope at metaphase I. The significance of these events, their similarities with, anddifferences from, comparable stages of meiosis in other plantsare discussed. Pteridium aquilinum (L.) Kuhn, bracken, meiosis, sporogenesis     

Evolution of the cytoplasmic organelles during female meiosis in Pisum sativum L.

In this paper we have traced the evolution of the cytoplasmic organelles in the female germinal cell of Pisum sativum L., from the beginning of meiosis to the early stages of the maturing megaspore, in order to correlate the morphological changes with the physiological aspects of megasporogenesis.A process of intense cytoplasmic vacuolation takes place in the megaspore mother cell (MMC) during prophase I, probably proceeding from the smooth endoplasmic reticulum and dictyosomes; it results in the formation of big vacuoles, which play a role in MMC polarization. By means of this polarization most plastids and mitochondria are incorporated into the functional megaspore at the end of meiosis.There are plastid and mitochondria cycles which consist of dedifferentiation followed by redifferentiation, During these cycles a transient morphology appears, called a cup-shaped form, which we interpret as an expression of low organelle activity.The wall of the MMC thickens throughout megasporogenesis and loses its plasmodesmata during middle prophase I. The ribosome population is reduced during prophase I and then restored during the early stages of the megaspore maturing process, as shown by the quantitative study that we have carried out. The nucleolar cytoplasmic bodies play a part in this restoring process. These bodies have a special morphology and appear to be originated from the activity of the nucleolar organizing region (NOR) during nucleolar disorganization in prophase I.We think that this cytoplasmic evolution is a response to nuclear genic recombination, in order to provide the most adequate expression of the zygote genome.Abbreviations EDTA ethylene-diamine-tetracetic acid - ER endoplasmic reticulum (SER: smooth ER) - MMC megaspore mother cell - NOR nucleolar organizing region - RNP ribonucleoproteins This work has been partially supported by the Comisión Asesora para la investigación Cientifica by Técnica Projects n^o 613/02 and 613/10     

The ribosome cycle,nucleoli, and cytoplasmic nucleoloids in the meiocytes ofLilium

Summary During the meiotic prophase in pollen mother cells ofLilium species the ribosome population of the cytoplasm diminishes to a minimum reached during the diplotene-diakinesis interval. This fall is associated with a drop in the ribonuclease-extractable RNA present in whole, fixed cells. A rise in RNA and in number of observable ribosomes follows during the meiotic mitoses and in the very early life of the spores. Bodies with the characteristics of nucleoli—here termed nucleoloids—are released into the cytoplasm at anaphase I and anaphase II, and it is suggested that these may represent the main mechanism through which the ribosome population of the cytoplasm is restored after the prophase elimination.     

Cellular organisation and differentiation of organelles in pre-meiotic rice anthers

Pre-meiotic cellular organisation of rice anthers has a great significance in pollen formation. We have used a combination of confocal laser and transmission electron microscopy (TEM) to characterise and differentiate organelles in pre-meiotic rice anthers. Along with the characteristic organelles in the cytoplasm the epidermal cells of the pre-meiotic rice anther are coated on their outer surface by a conspicuous bi-lamellate cuticle. Chloroplasts of the endothecium contain immature grana, thylakoids and also starch granules. These plastids clearly contain photosynthetic pigments as shown by autofluorescence in confocal microscope studies. Both confocal and TEM studies reveal clusters of mitochondria in the middle layer. The tapetum contains electron opaque ribosomes, bundles of mitochondria and plastids. The nuclei of the tapetum occupy a large volume of the cytoplasm indicating the onset of mitotic prophase. Intense Rhodamine 123 staining reveals that a major portion of the structurally indistinguishable organelles that were seen throughout the densely ribosomic cytoplasm of sporogenous cells are mitochondria.     

Ultrastructure of organelles during microsporogenesis inTillandsia pallidoflavens (Bromeliaceae)

InTillandsia pallidoflavens none of the organelles undergoes fundamental de- and redifferentiation during microsporogenesis. The plastids are amoeboid, exhibit complex internal structures and gradually start accumulating polysaccharides from meiotic prophase I onwards. These observations contradict reports for other taxa. The ultrastructure of mitochondria and dictyosomes, respectively, is more or less orthodox. The extensive ER, which is only poorly stained by standard methods was identified by image intensifiying techniques. The ribosomes are not only associated with the ER or occur as polyribosomes free in the cytoplasm, but can also form more or less dense clusters.     

Ultrastructure of meiosis in the mossRhynchostegium serrulatum I. Prophasic microtubules and spindle dynamics

Summary An extensive system of microtubules develops during meiotic prophase in the mossRhynchostegium serrulatum (Hedw.)Jaeg. &Sauerb. Development of the cytoskeleton can be traced to early prophase when the nucleus is acentric and the single plastid divides into four plastids. The cytoskeletal microtubules are associated with equidistant positioning of the four plastids at the distal tetrad poles and with migration of the nucleus to a central position in the sporocyte. The cytoskeleton, which interconnects plastids and encloses the nucleus, contributes to the establishment of moss sporocyte polarity. Just prior to metaphase I evidence of the prophase cytoskeleton is lost as the bipolar metaphase I spindle develops in association with discrete polar organizers located in opposite cleavage furrows between plastids.     

LIGHT AND ELECTRON MICROSCOPIC STUDIES OF THE FUSION CELL IN ASTEROCOLAX GARDNERI SETCH. (RHODOPHYTA,CERAMIALES)12

The fusion cell in Asterocolax gardneri Setch, is a large, multinucleate, irregularly-shaped cell resulting from cytoplasmic fusions of haploid and diploid cells. Subsequent enlargement takes place by incorporating adjacent gonimoblast cells. The resultant cell consists of two parts—a central portion of isolated cytoplasm, surrounded by an electron dense cytoplasmic barrier, and the main component of the fusion cell cytoplasm surrounding the isolated cytoplasm. The fusion cell contains many nuclei, large quantities of floridean starch, endoplasmic reticulum, and vesicles, but few mitochondria, plastids and dictyosomes. The endoplasmic reticulum forms vesicles that apparently secrete large quantities of extracellular mucilage which surrounds the entire carposporophyte. The isolated cytoplasm also is multinucleate but lacks starch and a plasma membrane. Few plastids, ribosomes and mitochondria are found in this cytoplasm. However, numerous endoplasmic reticulum cisternae occur near the cytoplasmic barrier and they appear to secrete material for the barrier. In mature carposporophytes, all organelles in the isolated cytoplasm have degenerated.     

Division polarity,development and configuration of microtubule arrays in bryophyte meiosis

Summary First and second division spindles and the three cell plates of moss meiosis are oriented in accordance with polarity established during meiotic prophase. Plastids are located at the second division poles and cytoplasmic infurrowing marks the planes along which the cytoplasm will cleave into four spores. Anaphase I spindles that terminate in two focal points of microtubules straddling opposite cleavage furrows reflect the unusual tetrahedral origin of the functionally bipolar spindle. The organelles (except for the plastids which remain in the four cytoplasmic lobes) are polarized in the first division equatorial region at the time of phragmoplast microtubule assembly and remain in a distinct band after microtubule disassembly. Prophasic spindles appear to be directly transformed into metaphase II spindles in the predetermined axes between mutually perpendicular pairs of plastids. Cell plates form by vesicle coalescence in the equatorial regions of the two sets of second division phragmoplasts at approximately the same time as a cell plate belatedly forms in the organelle band. The cytoplasmic markers (plastid migration, cytoplasmic lobing and infurrowing) that predict poles and cleavage planes in free cells lacking a preprophase band strongly strengthens the concept that division sites are capable of preserving preprogrammed signals that can be triggered later in the process of cell division.     

Subcellular aspects of differentiation of microspore mother cells ofNicotiana tabacum

Summary Differentiation of microspore mother cells inNicotiana starts with a period of active vesiculation in archesporial cells which is attended by a reduction of ribosome population. Vesiculation has been interpreted to cause cell reorganization and with their presumed hydrolytic functions the vesicles are reckoned to eliminate macromolecules associated with the sporophytic phase and result in the formation of cells capable of gametophytic functions. However, a part of cell cytoplasm is preserved in the form of membrane inclusions, which are bi- or multimembranous structures. Therefore, the rationale behind elimination and preservation of part of ribosomes remains to be understood. Another important feature of cell reorganization, the significance of which is quite elusive, is the dedifferentiation of organelles mitochondria and plastids. The former become electron dense and lose their cristae and the latter become cup-shaped and show regression of lamellae.     

Fine structure of spermatogonia and primary spermatocytes in rabbits

Summary The fine structure of rabbit Spermatogonia and primary spermatocytes in meiotic prophase has been studied with different methods of preparation, including a technique for acid phosphatase activity. The spermatogonial cytoplasm is rich in free ribosomes and containes moderate amounts of vesicular, smooth-surfaced endoplasmic reticulum and mitochondria, a simple Golgi-apparatus, some micropinocytotic vesicles, and occasional multivesicular bodies, vacuoles and dense bodies with acid phosphatase activity. The large type A Spermatogonia have a prominent nucleolus and their mitochondria sometimes form clusters with a dense intermitochondrial substance, similar to that in spermatocytes.The nucleus and cytoplasm of primary spermatocytes increase markedly in volume and density during meiotic prophase. The Golgi apparatus enlarges and becomes more differentiated and finally forms small proacrosome granules. The endoplasmic reticulum produces numerous small, mainly smooth vesicles and might also be the source of a new organelle: numerous piles of narrow cisternae with opaque contents. These piles disintegrate late in prophase. The mitochondria become aggregated in clusters with dense intermitochondrial substance and their internal structure is characterized by highly dilated cristae and small particles, interpreted as mitochondrial ribosomes, in the matrix. The role of these structures in the formation of new mitochondria is discussed. The clusters of mitochondria finally disperse and their cores of dense intermitochondrial substance, possibly containing ribonucleoprotein, coalesce into a large chromatoid body similar to that in spermatids. Micropinocytosis and a few lysosomes occur in most spermatocytes. The pachytene nuclei show prominent nucleoli and a distinct sex vesicle without any synaptinemal complex.The importance for spermatid differentiation of some events taking place in the cytoplasm of primary spermatocytes is emphasized.Financial support for this study was received from the Swedish Medical Research Council.     

Spermatogenesis in Tenebrio molitor (Tenebrionidae,Coleoptera): A Fine Structure and Anti-tubulin Immunofluorescence Study

Spermatogonia and both generations of spermatocytes of Tenebrio molitor possess conventional bipolar spindles with only few aster MTs. Spindles in metaphase spermatogonia are surrounded by fenestrated two-layered cisternae and do not contain intraspindle membranes. In metaphase spermatocytes, a spindle envelope is missing, but intraspindle membranes are abundant. Mitochondria form long threads lateral to the nucleus in prophase I of meiosis. The elongated mitochondria also align parallel to the spindle apparatus in prometaphase I. As a consequence, the spindles reside in a cage formed of mitochondria. This arrangement may guarantee proper bisection of the chondriome during division. Cells are tightly packed during spermatogonial divisions and in prophase I, but large intercellular spaces develop when the first meiotic spindle assembles. Then, cytoplasmic bridges which persist between the cells as a result of incomplete cytokinesis appear as slender tubes. Anti-tubulin immunofluorescence using an antibody against acetylated α-tubulin revealed intense acetylation throughout spermatogonial mitosis but a low degree of α-tubulin acetylation in meiotic spindles prior to telophase. This may indicate a high microtubule turnover in meiosis.     

葡萄果实发育过程中果肉细胞超微结构的观察

用透射电镜观察了“巨峰”葡萄(Vitis vinifera×V.labrusca)果实3个发育时期中果肉细胞超微结构的变化。果实第一次快速生长期的果肉细胞超微结构表现出物质和能量代谢旺盛的特点。缓慢生长期的果实虽外部形态平静少变,但果肉细胞超微结构表现出深刻的变化:细胞核形状特化为裂瓣状是最显著的特点;线粒体数目丰富;粗面内质网槽库膨大形成的囊泡富集,出现向液泡汇融和向质膜靠近的现象;质膜内陷;液泡膜完整。另外,原生质也出现一些降解的现象。但总体结构特点表明果肉细胞在此期处于十分活跃的物质周转代谢和信息交换过程中。果实第二次快速生长期果肉细胞超微结构表现出衰老降解的特点,但线粒体结构依然完整,数量仍然丰富,原生质膜也保持了很好的完整性,这似乎与维持第二次快速生长或成熟有关。     

The ultrastructure of Sorubim lima (Teleostei, Siluriformes, Pimelodidae) spermatogenesis: premeiotic and meiotic periods

The ultrastructure of Sorubim lima spermatogenesis during the premeiotic and meiotic periods was studied. Our observations showed that the germ cells in the cysts are connected by cytoplasmic bridges and the mitotic and meiotic divisions are slightly asynchronous. The first and the last spermatogonial generations differ in the cellular and nuclear volume, nucleolus, chromatin condensation, distribution, size, density, and shape of the mitochondria, presence of 'lamellae anulata', amount and dimension of the 'nuages', and movement of the centrioles. In addition to the nuclear prophase structures, the spermatocyte I shows changes in all other cellular organelles and elongated vesicles appear in the cytoplasm. The accentuated cytoplasmic density and thickened walled vesicles are morphological characteristics that differentiate spermatocytes II from the other germ cells in the cysts of Sorubim lima testis.     

Meiosis requires a translational positive loop where CPEB1 ensues its replacement by CPEB4

Meiotic progression is driven by the sequential translational activation of maternal messenger RNAs stored in the cytoplasm. This activation is mainly induced by the cytoplasmic elongation of their poly(A) tails, which is mediated by the cytoplasmic polyadenylation element (CPE) present in their 3′ untranslated regions. Although polyadenylation in prophase I and metaphase I is mediated by the CPE‐binding protein 1 (CPEB1), this protein is degraded during the first meiotic division. Thus, raising the question of how the cytoplasmic polyadenylation required for the second meiotic division is achieved. In this work, we show that CPEB1 generates a positive loop by activating the translation of CPEB4 mRNA, which, in turn, replaces CPEB1 and drives the transition from metaphase I to metaphase II. We further show that CPEB1 and CPEB4 are differentially regulated by phase‐specific kinases, generating the need of two sequential CPEB activities to sustain cytoplasmic polyadenylation during all the meiotic phases. Altogether, this work defines a new element in the translational circuit that support an autonomous transition between the two meiotic divisions in the absence of DNA replication.     

Differentiation and Ultrastructure of the Protophloem Sieve Elements of Minor Veins in the Maize Leaves: Changes in the Protoplast

Protophloem sieve element differentiation in the minor veins of the maize ( Zea mays L. ) leaves was first evidenced as an increase of the wall thickness, which began in the comers of the cell and then extended to other parts of the wall, and the appearance of long rough endoplasmic reticulum cistemae distributed throughout the cytoplasm, and then the presence of characteristic crystalloid inclusions within the plastids. As differentiation progressed, long cisternae of rough endoplasmic reticulum appeared to transform into shorter forms and eventually aggregated into small stacks, losing their ribosomes during the process. The nuclei degenerated, although frequently persisted until very late in differentiation the stages of maturation, as darkly stained amorphous aggregates surrounded by double nuclear envelope or only inner membrane of nuclear envelope. Subsequently, the nuclear envelope collapsed and became discontinuous. At the beginning of nuclear degeneration the perinuclear spaces were partly dilated and sometimes the outer nuclear envelope in the dilated portions then ruptured, and was accompanied by the disappearance of the cytoplasmic portion near it. During the peried of nuclear degeneration, in addition to the endoplasmic reticulum, plastids and mitochondria underwent structural modification, while components such as ribosomes, cytoplasmic ground substances, vacuoles and dictyosomes disintegrated and disappeared. At maturity, the surviving protoplasmic components, including plasmalemma, mitochondria, small stacked smooth endoplasmic reticulum and P-type plastids with crystalloids, became parietal in position. As differentiation of adjacent metaphloem sieve elements proceeded, the protoplasmic components of the mature protophloem sieve elements progresively degenerated and finally obliterated.     

Spermatogenesis and spermiogenesis in Ascaris lumbricoides Var. suum

Reorganization of the prophase I nucleus marks the beginning of the first meiotic division. A pair of centrioles is present at each pole at metaphase I and mitochondria are not observed in the spindle area. A chromosomal pellicle, which resembles a kinetochore plate but has no apparent association with microtubules, surrounds each autosome at metaphase I and II. The sex body lags behind the autosomes at anaphase I and segregates differentially to one daughter cell. Mitochondria and a pair of centrioles are present in the spindle during the second meiotic division. Localized condensation of chromatin and fusion of the condensed chromatin of the secondary spermatocyte telophase nucleus results in a compact spermatid nucleus. Loss of spermatid cytoplasm is effected by the ejection of a cytophore vesicle.     

The ultrastructure of wheat leaves I. Changes due to natural senescence and the effects of kinetin and ABA on detached leaves incubated in the dark

Summary Detached wheat leaves incubated in water in darkness rapidly lose starch and are ultrastructurally degenerate by 5 days. Chloroplast ribosomes disappear before cytoplasmic ribosomes and groups of wavy membranes can be detected in both mature and degenerating chloroplasts. Large lipid bodies appear in the cytoplasm and vacuole during senescence, and osmiophilic fibrils and deposits develop in the microbodies. Treatment with kinetin via the transpiration stream markedly delays the loss of starch and maintains, but does not increase, chloroplast and cytoplasmic ribosome populations. ABA accelerates degenerative changes as compared with the water treated control leaves but does not induce any specific ultrastructural effects.