Diphtheria toxin-mediated cell ablation in developing pollen: Vegetative cell ablation blocks generative cell migration

PROTOPLASMA ◽  
1995 ◽  
Vol 187 (1-4) ◽  
pp. 144-154 ◽  
Author(s):  
D. Twell
Keyword(s):  
Cell Migration ◽  
Diphtheria Toxin ◽  
Vegetative Cell ◽  
Generative Cell ◽  
Cell Ablation

Diphtheria toxin receptor–mediated conditional and targeted cell ablation in transgenic mice

10.1038/90795 ◽  
2001 ◽  
Vol 19 (8) ◽  
pp. 746-750 ◽  
Author(s):  
Michiko Saito ◽  
Takao Iwawaki ◽  
Choji Taya ◽  
Hiromichi Yonekawa ◽  
Munehiro Noda ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Diphtheria Toxin ◽  
Cell Ablation ◽  
Diphtheria Toxin Receptor ◽  
Toxin Receptor

scholarly journals Autoradiographic studies of DNA and histone synthesis in successive differentiation stages of pollen grain in Hyacinthus orientalis L.

2014 ◽  
Vol 50 (3) ◽  
pp. 367-380 ◽  
Author(s):  
Elżbieta Bednarska
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Vegetative Cell ◽  
Developmental Stages ◽  
Generative Cell ◽  
Pollen Grain ◽  
Cell Nuclei ◽  
Hyacinthus Orientalis ◽  
Histone Synthesis ◽  
Generative Cells

DNA and histone synthesis in five consecutive morphological stages of <em>Hyacinthus orientalis</em> L. pollen grain differentiation were studied autoradiographically. DNA synthesis was found to occur in both the generative and the vegetative cell. DNA replication in the generative cell took place when the generative cell was still adhered to the pollen grain wall but already devoid of callose wall. DNA synthesis in the generative cell slightly preceded that in the vegetative cell. Histones were synthesized in phase S of the generative and vegetative cell. In the generative cell histone synthesis also continued at a lower level after completion of DNA replication. In the developmental stages under study the nuclei of the generative cells were decidedly richer in lysine histones than vegetative cell nuclei.

Development of sperm cells in barley

1975 ◽  
Vol 53 (10) ◽  
pp. 1051-1062 ◽  
Author(s):  
David D. Cass ◽  
Ilana Karas
Keyword(s):  
Cell Membrane ◽  
Vegetative Cell ◽  
Generative Cell ◽  
Pollen Grains ◽  
Complete Separation ◽  
Pollen Wall ◽  
Vegetative Nucleus ◽  
Sperm Cells ◽  
Subsequent Stage ◽  
Microspore Stage

Ultrastructural events in barley sperm development were examined from the uninucleate microspore stage to establishment of two mature sperm cells in pollen grains. Microspore mitosis produces a vegetative nucleus and a naked generative cell, both embedded in vegetative cell cytoplasm. The generative cell membrane is enclosed by vegetative cell membrane. The generative cell, at first apparently unattached, becomes attached to the pollen wall and acquires a cell wall by centripetal vesicle accumulation. Wall formation may be complete at the time of generative cell karyokinesis; karyokinesis occurs while the generative cell is attached to the pollen wall. Cytokinesis of the generative cell is delayed. The subsequent stage is a binucleate, attached generative cell with a wall. Generative cell cytokinesis appears to involve formation of a partition between the two sperm nuclei. Eventual complete separation of the sperm cells occurs only after the two-celled derivative of the generative cell detaches from the pollen wall. Final stages in sperm cell separation are considered to result from degradation of the partitioning and surrounding wall, not from furrowing of a naked binucleate generative cell according to previous suggestions. Mature plastids were not observed in the generative cell or the sperms.

scholarly journals Ultrastructural study of maturing pollen in Arabidopsis thaliana (L.) Heynh. (Brassicaceae)

2014 ◽  
Vol 66 (2) ◽  
pp. 125-131 ◽  
Author(s):  
Krystyna Zając
Keyword(s):  
Arabidopsis Thaliana ◽  
Vegetative Cell ◽  
Generative Cell ◽  
Spherical Shape ◽  
Ultrastructural Changes ◽  
Lipid Bodies ◽  
Cell Cytoplasm ◽  
Cell Form ◽  
Microspore Stage ◽  
Pollen Stage

Ultrastructural changes in <em>Arabidopsis thaliana</em> pollen, between late microspore stage and mature pollen stage were described. When the generative cell was peeled off from the intine, it was of spherical shape and had all usual organelles with the exception of plastids. The cytoplasm transformation of the vegetative cell included an increase in the number of mitochondria and changes in the accumulation of starch and lipid bodies. The starch plastids were observed at the bicellular and early tricellular pollen stages and next starch was utilized during the maturation procces. The lipid bodies of the vegetative cell form a very regular sheath around the generative cell and then, around the sperm cells. Before anthesis the lipid bodies were dispersed within the whole vegetative cell cytoplasm.

scholarly journals Vestibular and Auditory Hair Cell Regeneration Following Targeted Ablation of Hair Cells With Diphtheria Toxin in Zebrafish

2021 ◽  
Vol 15 ◽  
Author(s):  
Erin Jimenez ◽  
Claire C. Slevin ◽  
Luis Colón-Cruz ◽  
Shawn M. Burgess
Keyword(s):  
Hair Cell ◽  
Inner Ear ◽  
Lateral Line ◽  
Hair Cells ◽  
Diphtheria Toxin ◽  
Adult Zebrafish ◽  
Hair Cell Regeneration ◽  
Cell Ablation ◽  
Aquatic Vertebrates

Millions of Americans experience hearing or balance disorders due to loss of hair cells in the inner ear. The hair cells are mechanosensory receptors used in the auditory and vestibular organs of all vertebrates as well as the lateral line systems of aquatic vertebrates. In zebrafish and other non-mammalian vertebrates, hair cells turnover during homeostasis and regenerate completely after being destroyed or damaged by acoustic or chemical exposure. However, in mammals, destroying or damaging hair cells results in permanent impairments to hearing or balance. We sought an improved method for studying hair cell damage and regeneration in adult aquatic vertebrates by generating a transgenic zebrafish with the capacity for targeted and inducible hair cell ablation in vivo. This model expresses the human diphtheria toxin receptor (hDTR) gene under the control of the myo6b promoter, resulting in hDTR expressed only in hair cells. Cell ablation is achieved by an intraperitoneal injection of diphtheria toxin (DT) in adult zebrafish or DT dissolved in the water for larvae. In the lateral line of 5 days post fertilization (dpf) zebrafish, ablation of hair cells by DT treatment occurred within 2 days in a dose-dependent manner. Similarly, in adult utricles and saccules, a single intraperitoneal injection of 0.05 ng DT caused complete loss of hair cells in the utricle and saccule by 5 days post-injection. Full hair cell regeneration was observed for the lateral line and the inner ear tissues. This study introduces a new method for efficient conditional hair cell ablation in adult zebrafish inner ear sensory epithelia (utricles and saccules) and demonstrates that zebrafish hair cells will regenerate in vivo after this treatment.

scholarly journals Ultrastructural transformations in the cytoplasm of differentiating Hyacinthus orientalis L. pollen cells

2014 ◽  
Vol 57 (2) ◽  
pp. 235-245 ◽  
Author(s):  
Elżbieta Bednarska
Keyword(s):  
Vegetative Cell ◽  
Generative Cell ◽  
Pollen Grain ◽  
Border Region ◽  
Ultrastructural Changes ◽  
Grain Development ◽  
Callose Wall ◽  
Highly Active ◽  
Hyacinthus Orientalis

The sequence of ultrastructural changes in the cytoplasm during the successive stages of pollen grain development in <em>Hyacinthus orientulis</em> pollen cells was studied. The cytoplasmic transformations of the generative cell included the elimination of plastids, increase in the number of mitochondria, assumption of a spindle shape with the aid of microtubules and the characteristic development of the vacuole system with the formation of so-called colored bodies. The cytoplasmic transformations of the generative cell encompassed changes in the plastids, which began to accumulate starch soon after the cell was formed, then released it shortly before anthesis, an increase in the number of mitochondria and an increase in the number of highly active dictyosomes just before anthesis. Changes in the structure of the border region between the differentiating pollen cells were associated mainly with the periodical appearance of a callose wall and the presence of lysosome-like bodies in the cytoplasm of the vegetative cell surrounding the generative cell. They arose soon after the disappearance of the callose wall and disappeared shortly before anthesis.

Elektronenmikroskopische Beobachtungen an der vegetativen Zelle im auskeimenden Pollenkorn von Oenothera hookeri Torr. et Grey

1963 ◽  
Vol 18 (12) ◽  
pp. 1092-1097 ◽  
Author(s):  
Lothar Diers
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Nuclear Envelope ◽  
Vegetative Cell ◽  
Generative Cell ◽  
Pollen Grain ◽  
Serial Sections ◽  
Lipid Inclusions ◽  
Intense Activity ◽  
Germinating Pollen

According to the intense activity of the vegetative cell in the germinating pollen grain, the cytoplasm shows a highly organized structure. Concerning the structure the vegetative cell differs strongly from the generative cell. In the vegetative cell the big nucleus shows a very lobed shape. Large invaginations of the cytoplasm into the nucleus can be frequently observed. Series of adjacent sections show that deep and flat vesicles which may often broaden to unusual large cisternae, extend through the vegetative plasm and form by interconnections a highly developed endoplasmic reticulum which is continuous with the nuclear envelope. The leucoplasts contain large starch grains and very few lamellae, in many sections only one lamella is visible. Sometimes, a process of a leucoplast deeply reaches into another leucoplast. In some leucoplasts and mitochondria there are concentric stripes which, according to serial sections, are the margins of invaginations of the cytoplasm or of another organell. In the numerous mitochondria the inner folds have the form of cristae, tubules are not so frequently seen. The edges of the flattened sacs of the Golgi - apparatus expand to vacuoles which seem to separate from the flattened cisternae. Typical for the vegetative plasm are numerous small vacuoles. Relatively large, ringshaped or uniform dark bodies are assumed to be lipid inclusions.

Investigating the function of follicular subpopulations during Drosophila oogenesis through hormone-dependent enhancer-targeted cell ablation

Development ◽  
2000 ◽  
Vol 127 (3) ◽  
pp. 573-583 ◽  
Author(s):  
D.D. Han ◽  
D. Stein ◽  
L.M. Stevens
Keyword(s):  
Gene Expression ◽  
Diphtheria Toxin ◽  
Follicle Cells ◽  
Specific Gene ◽  
Border Cells ◽  
Drosophila Oogenesis ◽  
Cell Ablation ◽  
Powerful Approach ◽  
Human Estrogen Receptor

Although it is known that the establishment of polarity during Drosophila oogenesis is initiated by signalling from the oocyte to the overlying follicle cells, much less is understood about the role of specific follicular subpopulations. One powerful approach for addressing this question, toxigenic cell ablation of specific subpopulations, has not previously been applicable to studying follicular subpopulations because many of the genes and Gal4 enhancer trap insertions that are expressed in the ovary are also expressed at earlier times in development. To overcome this problem, we have utilized a fusion protein between Gal4 and the human estrogen receptor to achieve hormone-dependent, tissue-specific gene expression of UAS-linked transgenes in flies. We used this system to study the role of the polar subpopulations of follicle cells during oogenesis by expressing within them a modified form of diphtheria toxin that causes cell death. Our results confirmed previous functions ascribed to these cells, and also demonstrated a previously undescribed role for the border cells in facilitating the migration of the anterior Fasciclin III-expressing polar pair cells to the edge of the oocyte.

The Ultrastructure and Ontogeny of Pollen in Helleborus Foetidus L

1972 ◽  
Vol 11 (1) ◽  
pp. 111-129
Author(s):  
P. ECHLIN
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Vegetative Cell ◽  
Generative Cell ◽  
Ultrastructural Level ◽  
Mitotic Division ◽  
Tube Formation ◽  
Pollen Grain ◽  
Granular Body ◽  
Development Proceeds

The final stages of Helleborus pollen-grain ontogeny, which culminate in maturation and germination of the grain, have been investigated at the ultrastructural level. Following the deposition of primary and secondary exine, and during the early stages of intine formation, the microspore passes through a vacuolate phase, in which the cytoplasm appears devoid of most organelles other than the prominent nucleus. The formation of the vacuole results in the displacement of the nucleus to one side of the pollen grain. The vacuole quickly disappears and a number of organelles reappear in the cytoplasm, in particular the dictyosomes and strands of endoplasmic reticulum, with associated grey bodies. Following mitotic division of the pollen grain, the first signs of the generative cell wall appear as a pair of tightly appressed unit membranes in the narrow strand of cytoplasm separating the two newly formed generative and vegetative nuclei. As development proceeds, the space between the two membranes gradually fills with an electron-transparent material similar to the substance found in the numerous dictyosome-derived vesicles which, together with the endoplasmic reticulum, are both closely associated with the developing cell wall. The generative cell wall fuses with the cellulosic intine, which has gradually increased in amount during these stages, and the cell division is complete. The smaller generative cell contains a prominent nucleus and a small amount of cytoplasm devoid of plastids and most other organelles. The larger vegetative cell also contains a prominent nucleus and a large amount of cytoplasm containing amyloplasts, mitochondria, dictyosomes and endoplasmic reticulum, and abundant ribosomes, many of which are in a polysome configuration. The final stages in development are characterized by a progressive decrease in the amount of starch in the vegetative cell and an increase in the size of grey bodies, many of which are invested in multilayered shrouds of endoplasmic reticulum. The generative cell wall disappears and a multivesicular/granular body gradually appears at the periphery of the pollen grain. The granular-vesicular material, which is formed from the dictyosomes and/or the degenerating plastids, is thought to represent metabolic reserves necessary for pollen-tube formation. One or more pollen tubes emerge from the apertural sectors of the pollen grain, and maturation of the grain is complete.

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