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Biomedical subjects

E Robbins

Publications and source records attributed to E Robbins.

At least 73 records · Page 4Linked to original sources
PubMed1970

Morphologic changes accompanying senescence of cultured human diploid cells.

The lysosomes of serially propagated human fibroblasts gradually transform to residual bodies which increase in number and size, and show progressive degenerative changes. There is an accompanying, and less regular, decrease in the number of cytoplasmic polyribosomes and an increased number of glycogen particles. The onset of these morphologic alterations occurs shortly after culture initiation and precedes any marked decrease in the rate of cellular growth; however, in their extreme form these changes may be related to the ultimate cessation of cellular multiplication ("senescence"). The lysosomal changes were seen only in those cell strains which eventually showed senescence, and were absent or minimal either in cell lines which can be propagated indefinitely ("spontaneous" and viral transformants, cancer cells), or in skin sections from aging subjects.

Aging↗
PubMed1970

Iron: its intracellular localization and possible role in cell division.

When HeLa cells are fixed with ethanol, extracted with 0.1 N HCl, and incinerated at 540 degrees C, all organic constituents and all ions studied are removed with the exception of iron. The gross outlines of cell structures are preserved and high concentrations of residual ash in interphase nucleoli and mitotic chromosomes suggest that there may be a shift in iron salts during the cell cycle. Experiments with cells isotopically labeled in proteins, nucleic acids, lipids, and polysaccharides indicate that the iron is bound to a polysaccharide. Addition of iron chelating agents to living cells causes a selective inhibition of DNA synthesis. These data suggest that iron may play a crucial role in the mitotic process.

Calcium↗
PubMed1970

Comparison of mitotic phenomena and effects induced by hypertonic solutions in HeLa cells.

Interphase HeLa cells exposed to solutions that are 1.6 x isotonic manifest a series of morphological transformations, several of which grossly resemble those which occur when untreated cells enter prophase. These include chromosome condensation with preferential localization at the nuclear envelope and nucleolus, ruffling of the nuclear envelope, and polyribosome breakdown. The nucleolus loses its fibrous component and appears diffusely granular. At 2.8 x isotonicity the nuclear envelope is selectively dispersed although other membranes show morphological alterations also. The characteristic transitions of the lysosomes, Golgi complex, and microtubules seen in normal mitosis do not occur during hypertonic treatment. All the changes induced with hypertonic solutions are rapidly reversible, and the nucleus particularly goes through a recovery phase which bears some similarity to that of the telophase nucleus. The prophase-like condensation of the chromatin following exposure of the intact cell to hypertonic medium cannot be reproduced on an ultrastructural level in the isolated nucleus with any known variation in salt concentration, suggesting significant modifications of the nuclear contents during isolation. In addition to these morphological responses, hypertonic solutions also markedly and reversibly depress macromolecular synthesis. The polyribosome disaggregation that results from exposure to hypertonic solutions may be partially prevented by prior exposure to elevated Mg(++) concentrations; this same ion is also partially effective in preventing the polyribosome breakdown which normally occurs as cells enter mitosis.

Cell Nucleus↗
PubMed1970

Absence of translational control of histone synthesis during the HeLa cell life cycle.

The cell-free synthesis of histone-like polypeptides has been achieved using a selected class of small polyribosomes as the only particulate fraction. This synthesis is prevented if the deoxyribonucleic acid (DNA) inhibitor, cytosine arabinoside, is added to the cells prior to disruption, and it is not detected when the cytoplasm used is derived from postmitotic (G(1)) cells. When the 100,000 g supernate from pure metaphase populations was compared with that from S phase cells, the cell-free synthesis of histone-like polypeptides in the presence of S phase polyribosomes remained unchanged. These data suggest that, except for the histone messenger RNA-ribosome complex, the cytoplasmic factors requisite for histone synthesis are present throughout the cycle, and that the shut-off of this synthesis is not under translational control.

Carbon Isotopes↗
PubMed1970

RNA synthesis in HeLa cells. Pattern in hypertonic medium and its similarity to synthesis during G2-prophase.

Interphase HeLa cells manifest a stepwise shutoff of RNA synthesis when the tonicity of the extracellular medium is gradually increased. Synthesis of heterogeneous nuclear RNA is most sensitive and is selectively inhibited at 1.5 times isotonicity (450 milliosmols/liter), while 45S ribosomal RNA synthesis is not affected significantly below 2.0 times isotonicity. Transfer RNA synthesis is least sensitive to increased osmolarity and is not completely inhibited until the electrolyte concentration of the medium is elevated to 2.8 times isotonicity. Although the transcription and methylation of 45S ribosomal precursor is unaffected at 1.5 times isotonicity, there is pronounced impairment of its processing into 32S and 18S RNA. Using a refined cell synchronization technique, we have been able to compare these effects of hypertonicity with the shutoff of RNA synthesis which occurs during the G(2)-prophase interval of the cell division cycle. In this case, as with random cells in hypertonic medium, a selective inhibition of heterogeneous nuclear RNA synthesis and slowed processing of 45S ribosomal RNA were found, whereas synthesis of 45S and transfer RNA continued unabated throughout G(2)-prophase. While it is known that RNA synthesis essentially ceases during metaphase, we have noted that transfer RNA synthesis continues in metaphase at 10-15% of the interphase rate, which is of particular interest in view of the relative resistance of this species to hypertonicity. The close correlation between the patterns of cessation of RNA synthesis at mitosis and during exposure to hypertonic medium supports our earlier contention that alteration of intracellular electrolyte levels provides a useful model for studying the mechanism of mitosis.

Carbon Isotopes↗
PubMed1969

Persistence of messenger RNA through mitosis in HeLa cells.

The decrease in protein synthesis which occurs in mammalian cells during cell division is associated with significant disaggregation of polyribosomes. For determining whether messenger RNA survives this disaggregation, the reformation of polyribosomes was investigated in synchronized HeLa cells as they progressed from metaphase into interphase in the presence of 2 microg/ml Actinomycin D. The persistence of messenger during cell division was evidenced by: (1) a progressive increase in the rate of protein synthesis in both treated and untreated cells for 45 min after metaphase; (2) reformation of polyribosomes, as determined by both sucrose gradients and electron microscopy, within 30 min after the addition of Actinomycin D to metaphase cells; (3) the persistence of approximately 50% of the rapidly labeled nonribosomal RNA which had associated with polyribosomes just before metaphase; (4) the resumption of synthesis, following cell division, of 6 selected peptides in Actinomycin-treated cells.

Centrifugation, Density Gradient↗
PubMed1969

Ultrastructural changes in the mitotic apparatus at the metaphase-to-anaphase transition.

As the metaphase HeLa cell approaches anaphase, pericentriolar spindle tubules fragment and become encapsulated by a unit membrane. By early anaphase, the encapsulated forms appear to have expanded, giving rise to polar spherical aggregates. Some of these elements show ribosomes on their bounding membrane, and some of them localize on the condensed chromatin during reformation of the nuclear membrane. It thus is suggested that these elements are newly derived cisternae of the endoplasmic reticulum (ER). Similar transformations are seen in later anaphase in the interzonal region, and it may be that the ER serves as a storage depot for some fraction of depolymerized microtubules. The time and location of the pericentriolar transitions are consistent with their being intimately involved in the mechanics of chromosome separation.

Endoplasmic Reticulum↗
PubMed1968

The centriole cycle in synchronized HeLa cells.

Progression of the HeLa cell through its life cycle is accompanied by centriolar replication and pericentriolar changes that are in synchrony with DNA synthesis and mitosis. The first signs of preparation for replication occur during G(1) at which time the two orthogonal centrioles separate. Replication by budding begins at/or near the initiation of DNA synthesis and is completed by G(2). Pericentriolar changes which probably are causally related to spindle tubule formation occur at this time and include the appearance of vesicles, electron-opaque bodies, and an amorphous pericentriolar halo. These phenomena begin to disappear by late prophase, and the remainder of mitosis manifests decreasing centriolar and pericentriolar activity.

Carbon Isotopes↗