Mitonucleons formed during differentiation of Ishikawa endometrial cells generate vacuoles that elevate monolayer syncytia: Differentiation of Ishikawa domes, Part 1

10.7287/peerj.preprints.1728 ◽

2016 ◽

Cited By ~ 1

Author(s):

Honoree Fleming

Keyword(s):

Cell Death ◽

Structural Changes ◽

Apical Membrane ◽

Signet Ring Cells ◽

Double Membrane ◽

Endometrial Cells ◽

Endometrial Epithelial Cell ◽

Signet Ring ◽

Endogenous Biotin ◽

Cell Profile

In 1998, we published a paper (Fleming et.al, 1998) describing some aspects of Ishikawa endometrial epithelial cell differentiation from monolayer cells into cells forming fluid-filled hemispheres called domes. The process begins with the dissolution of membranes within discrete regions of the monolayer. Nuclei from fused cells aggregate and endogenous biotin in particulate structures assumed to be mitochondria increase throughout the resulting syncytium. Endogenous biotin is also the distinguishing feature of a membrane that surrounds aggregates of multiple nuclei in a structure called a mitonucleon. The current paper includes additional observations on structural changes accompanying Ishikawa differentiation. Vacuoles form in the heterochromatin of the mitonucleon and within the biotin-containing double membrane surrounding heterochromatin. With the formation of vacuoles, the mitonucleon can be seen to rise along with the apical membrane of the syncytium in which it formed. The small vacuoles that form within the heterochromatin result in structures similar to “cells with optically clear nuclei” found in some cancers. The second larger vacuole that forms within the membrane surrounding the heterochromatin transforms the cell profile to one that resembles “signet ring” cells also observed in some cancers. Eventually the membrane surrounding the massed heterochromatin, generated three to four hours earlier, is breached and previously aggregated nuclei disaggregate. During this process heterochromatin in the mitonucleons undergoes changes usually ascribed to cells undergoing programmed cell death such as pyknosis and DNA fragmentation (Fleming, 2016b). The cells do not die, instead chromatin filaments appear to coalesce into a chromatin mass that gives rise to dome-filling nuclei by amitosis during the final three to four hours of the 20 hour differentiation (Fleming, 2016c).

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Pyknotic chromatin in mitonucleons elevating in syncytia undergo karyorhhexis and karyolysis before coalescing into an irregular chromatin mass: Differentiation of Ishikawa Domes, Part 2

10.7287/peerj.preprints.1729v1 ◽

2016 ◽

Cited By ~ 1

Author(s):

Honoree Fleming

Keyword(s):

Cell Death ◽

Epithelial Cells ◽

Programmed Cell Death ◽

Apical Membrane ◽

Driving Forces ◽

Double Membrane ◽

Membrane Staining ◽

Fragmented Dna ◽

Endogenous Biotin ◽

Irregular Mass

Pyknosis, karyorrhexis and karyolysis, harbingers of programmed cell death in many systems, appear to be driving forces that transform Ishikawa monolayer epithelial cells into differentiated dome cells. The heterochromatin affected by these process is contained in multiple nuclei aggregated in the syncytia that form when Ishikawa monolayers are stimulated to differentiate (Fleming, 2016a). The nuclear aggregates are enveloped in a double membrane staining for the endogenous biotin in mitochondrial carboxylases. The structure called a mitonucleon becomes vacuolated, along with the heterochromatin it envelops, and this structure elevates with the apical membrane of the syncytium 6 to 8 hours into the 20 hour differentiation, becoming increasingly pyknotic. This phase of the differentiation comes to an end when the mitonucleon membranes are breached and nuclei emerging from the aggregated state can be seen to fragment explosively. Fragmented DNA associates with an array of microtubules, filling the large central clearing of the predome. Some chromatin remains unfragmented and can be seen of the edges of the predome clearing. Cell death does not occur. Instead, the fragmented DNA coalesces into an irregular mass within the apical and basal membranes of the predome under which fluid has been accumulating. From the chromatin sheet, nuclei emerge amitotically as described in Part 3 of this series (Fleming, 2016c).

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Pyknotic chromatin in mitonucleons elevating in syncytia undergo karyorhhexis and karyolysis before coalescing into an irregular chromatin mass: Differentiation of Ishikawa Domes, Part 2

10.7287/peerj.preprints.1729 ◽

2016 ◽

Author(s):

Honoree Fleming

Keyword(s):

Cell Death ◽

Epithelial Cells ◽

Programmed Cell Death ◽

Apical Membrane ◽

Driving Forces ◽

Double Membrane ◽

Membrane Staining ◽

Fragmented Dna ◽

Endogenous Biotin ◽

Irregular Mass

Pyknosis, karyorrhexis and karyolysis, harbingers of programmed cell death in many systems, appear to be driving forces that transform Ishikawa monolayer epithelial cells into differentiated dome cells. The heterochromatin affected by these process is contained in multiple nuclei aggregated in the syncytia that form when Ishikawa monolayers are stimulated to differentiate (Fleming, 2016a). The nuclear aggregates are enveloped in a double membrane staining for the endogenous biotin in mitochondrial carboxylases. The structure called a mitonucleon becomes vacuolated, along with the heterochromatin it envelops, and this structure elevates with the apical membrane of the syncytium 6 to 8 hours into the 20 hour differentiation, becoming increasingly pyknotic. This phase of the differentiation comes to an end when the mitonucleon membranes are breached and nuclei emerging from the aggregated state can be seen to fragment explosively. Fragmented DNA associates with an array of microtubules, filling the large central clearing of the predome. Some chromatin remains unfragmented and can be seen of the edges of the predome clearing. Cell death does not occur. Instead, the fragmented DNA coalesces into an irregular mass within the apical and basal membranes of the predome under which fluid has been accumulating. From the chromatin sheet, nuclei emerge amitotically as described in Part 3 of this series (Fleming, 2016c).

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Polyploid monolayer Ishikawa endometrial cells form unicellular hollow spheroids capable of migration

10.7287/peerj.preprints.26793 ◽

2018 ◽

Author(s):

Honoree Fleming

Keyword(s):

Petri Dish ◽

Signet Ring Cell ◽

Polyploid Cell ◽

Central Vacuole ◽

Signet Ring Cells ◽

Endometrial Cells ◽

Short Period ◽

The Family ◽

Signet Ring ◽

Ring Cell

The results in this paper demonstrate that Ishikawa endometrial monolayer cells become multinucleated by a process of nuclear “donation” from neighboring cells. As the resulting polyploid cell detaches from the colony in which it was formed, it is possible to detect mitonucleon(s) in the center of the cell. The mitonucleon is a transient mitochondrial superstructure surrounding aggregated chromatin (Fleming et al. 1998) with characteristics of the family of mitochondrial superstructures that are sometimes called spheroids or cup-shaped mitochondria (Fleming, 2016a). As was recently demonstrated gas vacuoles form within mitonucleons (Fleming, 2018). In the free-floating single cell, the retained gas creates a central vacuole, and the cell becomes a spheroid that floats above the monolayer. It resembles a “signet ring cell” in being characterized by a central vacuole and chromatin compressed against the vacuole membrane. The resulting structure is a spheroids that is hollow and unicellular, albeit polyploid. But whereas signet ring cells are assumed to be undergoing apoptosis, that is not the case for unicellular spheroids. Complete spheres with chromatin and cytosolic cell contents compressed against the cell membrane can be found floating independently above Ishikawa monolayers. When an isolated sphere settles back onto the surface of the petri dish, it is possible to observe dissipating gas bubbles within the now flattened sphere for a short period of time. When the gas is discharged the resulting cell looks like a typical giant polyploid cell.

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Polyploid monolayer Ishikawa endometrial cells form unicellular hollow spheroids capable of migration

10.7287/peerj.preprints.26793v1 ◽

2018 ◽

Author(s):

Honoree Fleming

Keyword(s):

Petri Dish ◽

Signet Ring Cell ◽

Polyploid Cell ◽

Central Vacuole ◽

Signet Ring Cells ◽

Endometrial Cells ◽

Short Period ◽

The Family ◽

Signet Ring ◽

Ring Cell

The results in this paper demonstrate that Ishikawa endometrial monolayer cells become multinucleated by a process of nuclear “donation” from neighboring cells. As the resulting polyploid cell detaches from the colony in which it was formed, it is possible to detect mitonucleon(s) in the center of the cell. The mitonucleon is a transient mitochondrial superstructure surrounding aggregated chromatin (Fleming et al. 1998) with characteristics of the family of mitochondrial superstructures that are sometimes called spheroids or cup-shaped mitochondria (Fleming, 2016a). As was recently demonstrated gas vacuoles form within mitonucleons (Fleming, 2018). In the free-floating single cell, the retained gas creates a central vacuole, and the cell becomes a spheroid that floats above the monolayer. It resembles a “signet ring cell” in being characterized by a central vacuole and chromatin compressed against the vacuole membrane. The resulting structure is a spheroids that is hollow and unicellular, albeit polyploid. But whereas signet ring cells are assumed to be undergoing apoptosis, that is not the case for unicellular spheroids. Complete spheres with chromatin and cytosolic cell contents compressed against the cell membrane can be found floating independently above Ishikawa monolayers. When an isolated sphere settles back onto the surface of the petri dish, it is possible to observe dissipating gas bubbles within the now flattened sphere for a short period of time. When the gas is discharged the resulting cell looks like a typical giant polyploid cell.

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Primary thymic adenocarcinoma with signet ring cells: An unexpected autopsy finding

10.26226/morressier.578f37fdd462b8028d890452 ◽

2016 ◽

Author(s):

João Fraga

Keyword(s):

Autopsy Finding ◽

Signet Ring Cells ◽

Thymic Adenocarcinoma ◽

Signet Ring

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Gonadotrophins modulate cell death-related genes expression in human endometrium

Hormone Molecular Biology and Clinical Investigation ◽

10.1515/hmbci-2019-0074 ◽

2020 ◽

Vol 41 (2) ◽

Author(s):

Sandro Sacchi ◽

Paola Sena ◽

Chiara Addabbo ◽

Erika Cuttone ◽

Antonio La Marca

Keyword(s):

Cell Death ◽

Quantitative Polymerase Chain Reaction ◽

Hypoxia Inducible Factor ◽

Endometrial Cells ◽

Microtubule Associated Proteins ◽

Genes Expression ◽

Associated Proteins ◽

Polymerase Chain ◽

Cytochrome P450 Family ◽

Apoptosis Marker

AbstractBackgroundGonadotrophins exert their functions by binding follicle-stimulating hormone receptor (FSHR) or luteinizing hormone and human chorionic gonadotropin receptor (LHCGR) present on endometrium. Within ovaries, FSH induces autophagy and apoptosis of granulosa cells leading to atresia of non-growing follicles, whereas hCG and LH have anti-apoptotic functions. Endometrial cells express functioning gonadotrophin receptors. The objective of this study was to analyze the effect of gonadotrophins on physiology and endometrial cells survival.Materials and methodsCollected endometria were incubated for 48 or 72 h with 100 ng/mL of recombinant human FSH (rhFSH), recombinant human LH (rhLH) or highly purified hCG (HPhCG) alone or combined. Controls omitted gonadotrophins. The effect of gonadotrophins on cytochrome P450 family 11 subfamily A polypeptide 1 (CYP11A1), hypoxia inducible factor 1α (HIF1A), and cell-death-related genes expression was evaluated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Immunohistochemistry for microtubule-associated proteins 1A/1B light chain 3B (MAP1LC3B) and apoptotic protease activating factor 1 (APAF-1) was performed.ResultsGonadotrophins are able to modulate the endometrial cells survival. FSH induced autophagy and apoptosis by increasing the relative expression of MAP1LC3B and FAS receptor. In FSH-treated samples, expression of apoptosis marker APAF-1 was detected and co-localized on autophagic cells. hCG and LH does not modulate the expression of cell-death-related genes while the up-regulation of pro-proliferative epiregulin gene was observed. When combined with FSH, hCG and LH prevent autophagy and apoptosis FSH-induced.ConclusionsDifferent gonadotrophins specifically affect endometrial cells viability differently: FSH promotes autophagy and apoptosis while LH and hCG alone or combined with rhFSH does not.

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Suppression of cell proliferation and programmed cell death by dexamethasone during postnatal lung development

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00406.2000 ◽

2002 ◽

Vol 282 (3) ◽

pp. L477-L483 ◽

Cited By ~ 31

Author(s):

Cédric Luyet ◽

Peter H. Burri ◽

Johannes C. Schittny

Keyword(s):

Cell Proliferation ◽

Cell Death ◽

Programmed Cell Death ◽

Lung Development ◽

Structural Changes ◽

Tissue Transglutaminase ◽

Lung Parenchyma ◽

Lung Maturation ◽

Morphological Criteria ◽

Postnatal Lung Development

Prematurely born babies are often treated with glucocorticoids. We studied the consequences of an early postnatal and short dexamethasone treatment (0.1–0.01 μg/g, days 1–4) on lung development in rats, focusing on its influence on peaks of cell proliferation around day 4 and of programmed cell death at days 19–21. By morphological criteria, we observed a dexamethasone-induced premature maturation of the septa ( day 4), followed by a transient septal immatureness and delayed alveolarization leading to complete rescue of the structural changes. The numbers of proliferating (anti-Ki67) and dying cells (TdT-mediated dUTP nick end labeling) were determined and compared with controls. In dexamethasone-treated animals, both the peak of cell proliferation and the peak of programmed cell death were reduced to baseline, whereas the expression of tissue transglutaminase (transglutaminase-C), another marker for postnatal lung maturation, was not significantly altered. We hypothesize that a short neonatal course of dexamethasone leads to severe but transient structural changes of the lung parenchyma and influences the balance between cell proliferation and cell death even in later stages of lung maturation.

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