Prematurely condensed chromosomes of dividing and non-dividing cells in aging human cell cultures.

Chromosomes of dividing and non-dividing aging cells were examined by fusing senescent WI38 cells with mitotic HeLa cells to induce premature chromosome condensation (PCC). Exposure of the WI38 cells to ³H-thymidine 48 h prior to fusion allowed autoradiographic identification of cells that did not synthesize DNA (non-dividing cells). Ninety-six percent of the non-dividing cells, diploid or tetraploid, induced into PCC had single chromatids and were therefore blocked in the G1 phase of the cell cycle. Anomalous centromeric pairing of chromatids was noted in the remaining 4% of the non-dividing cells. Typical G2 configurations (double chromatids) were observed only among labeled (dividing) cells. The efficiency of PCC induction was independent of culture age. In addition, the efficiency of PCC induction was independent of phase in the cell cycle, as shown by comparison of observed frequencies with expected frequencies.     

Some histochemical observations on Gaucher cells

Plastic-embedded bone marrow biopsies from four patients with Gaucher's disease have been studied histochemically. Concanavalin A (ConA) was found to bind to cytoplasmic inclusions of Gaucher cells; the binding was prevented by lipid extraction or beta-glucosidase digestion. This suggests that glucocerebrosides stored in Gaucher cells are responsible for ConA binding; ConA staining combined with lipid extraction and beta-glucosidase digestion tests may be taken as a tool for the demonstration of Gaucher's cerebrosides of possible practical importance in diagnosis and investigation of Gaucher's disease. An excess of vic-glycol groups with respect to ConA binding-sugar residues and not extractable by lipid solvents are demonstrable in Gaucher cells. Vic-glycols appear to be regularly arranged at the electron microscopy level within Gaucher cell lysosomes along typical Gaucher "tubules", where some kind of interaction between lipid and protein should occur. Acid phosphatase might be one protein species involved in such interaction.     

Further observations on dividing and non-dividing cnidoblasts in the regenerating isolated gastrodermis of Hydra

Summary Cnidoblasts derived from the dedifferentiation of gland cells in the regenerating isolated gastrodermis of Hydra are capable of nuclear and cytoplasmic division. The daughter cell containing the nematocyst apparently develops normally. The fate of the other daughter cell remains obscure, but it is believed that it is also capable of developing a nematocyst. Only a single bi-nucleated cnidoblast was observed and it was in the process of degeneration. It is suggested that at least in the present system, cnidoblasts are derived not only from interstitial cells but also from pre-existing cnidoblasts.This investigation was supported by the National Science Foundation Grant No. GB 8384.With the technical assistance of Linda Bookman.     

DNA repair mechanisms in dividing and non-dividing cells

DNA damage created by endogenous or exogenous genotoxic agents can exist in multiple forms, and if allowed to persist, can promote genome instability and directly lead to various human diseases, particularly cancer, neurological abnormalities, immunodeficiency and premature aging. To avoid such deleterious outcomes, cells have evolved an array of DNA repair pathways, which carry out what is typically a multiple-step process to resolve specific DNA lesions and maintain genome integrity. To fully appreciate the biological contributions of the different DNA repair systems, one must keep in mind the cellular context within which they operate. For example, the human body is composed of non-dividing and dividing cell types, including, in the brain, neurons and glial cells. We describe herein the molecular mechanisms of the different DNA repair pathways, and review their roles in non-dividing and dividing cells, with an eye toward how these pathways may regulate the development of neurological disease.     

Some observations on the structure, cation content and possible evolutionary status of dinoflagellate chromosomes

SICEE, D. C., 1984. Some observations on the structure, cation content and possible evolutionary status of dinoflagellate chromosomes. Dinoflagellate chromosomes have a well-ordered structure, as observed in living cells, glutaraldehyde/osmium tetroxide-fixed cells, ultrathin cryosections and freeze-etch preparations. It is suggested that the stabilization of this chromatin in the living cell is largely mediated by divalent cations, acting as bridging molecules between the DNA superstructure and the protein matrix. Studies using X-ray micro-analysis and autoradiography have shown that these chromosomes have high levels of bound Ca and transition metals, and that these are associated with both the DNA and surrounding proteins.
The organization and stabilization of chromatin in dinoflagellate chromosomes is quite different from that of the cells of other eukaryotes, but shows some resemblance to the dispersed chromatin of bacteria. The evolution of dinoflagellate chromosomes from a prokaryote-like ancestral genome is attributed to two main factors–the retention of a primitive cationic non-histone stabilization system, and a pronounced evolutionary trend towards high DNA values. On this theory, dinoflagellate chromosomes are phylogenetically distinct from all other eukaryote chromosomes, and provide a separate evolutionary route for the attainment of high DNA levels and increased cell size.     

Mismatch repair as a source of mutations in non-dividing cells

This paper describes a mechanism which permits somatic cells to generate random mutations in the complete absence of cell proliferation. The mechanism itself is remarkably simple, involving a well-known cellular process (mismatch repair or MMR) which is primarily associated with mutation avoidance, but which is also capable of generating mutations when circumstances are not ideal for avoidance. When MMR operates in its so-called methylation-instructed mode to remove mismatches from newly-replicated portions of genomic DNA, it does so in a way which serves to minimize mutation yields. By contrast, when MMR operates in a non-instructed or randomly-templated way to remove mismatches from DNA molecules, it does so without distinguishing between the two strands of DNA that contain the mismatched bases. Randomly-templated mismatch repair (RT-MMR) therefore generates new and complete mutations whenever it removes the correct bases from either base-pair mismatches or frameshift mispairs and replaces them without incorrect bases or sequences. Wider recognition of the existence of this mechanism — and especially of its proclivity for mutation generation when it is operating in non-dividing cells—should help us to develop a better understanding of a number of important biological phenomena, and may be of particular value in our attempts to explain the origins of many human cancers.     

Cortical microtubule labeling: Insight of AFH14 in non-dividing cells

We recently reported that AFH14 participated in microtubule and actin filament interaction in cell division, and the AFH14 (FH1FH2) was important to the directly binding activity of microtubules and microfilaments. To preliminarily understand the function and localization of AFH14 in non-dividing cells, we overexpressed FH1FH2-RFP in onion epidermal cells, and found a fluorescence labeled filamentous network. The result of double labeling with different cytoskeleton reporter proteins indicated that FH1FH2-RFP co-localized with cortical microtubules. Treatment of cells expressing FH1FH2-RFP with cytoskeleton disrupting drugs confirms that FH1FH2-RFP binds to microtubules. Moreover, the binding of FH1FH2-RFP to microtubules were revealed to be dynamic by fluorescence recovery after photobleaching (FRAP) experiment. Time-lapse confocal microscopy showed that FH1FH2-RFP could display a dynamics similar to the microtubule dynamic instability. These data suggest that FH1FH2 domain may lead AFH14 function on cortical microtubules in non-dividing cells, and FH1FH2-RFP may be utilized as a microtubule reporter protein in living onion epidermal cells.Key words: cortical microtubule, AFH14, non-dividing cell, microtubule dynamics, FRAP