Search PubMed⌕ Search

Biomedical subjects

I M Gelfand

Publications and source records attributed to I M Gelfand.

At least 37 records · Page 2Linked to original sources
PubMed1995

Analysis of the relation between the sequence and secondary and three-dimensional structures of immunoglobulin molecules.

Methods of structural and statistical analysis of the relation between the sequence and secondary and three-dimensional structures are developed. About 5000 secondary structures of immunoglobulin molecules from the Kabat data base were predicted. Two statistical analyses of amino acids reveal 47 universal positions in strands and loops. Eight universally conservative positions out of the 47 are singled out because they contain the same amino acid in > 90% of all chains. The remaining 39 positions, which we term universally alternative positions, were divided into five groups: hydrophobic, charged and polar, aromatic, hydrophilic, and Gly-Ala, corresponding to the residues that occupied them in almost all chains. The analysis of residue-residue contacts shows that the 47 universal positions can be distinguished by the number and types of contacts. The calculations of contact maps in the 29 antibody structures revealed that residues in 24 of these 47 positions have contacts only with residues of antiparallel beta-strands in the same beta-sheet and residues in the remaining 23 positions always have far-away contacts with residues from other beta-sheets as well. In addition, residues in 6 of the 47 universal positions are also involved in interactions with residues of the other variable or constant domains.

Amino Acid Sequence↗
PubMed1995

Dynamics of active lamellae in cultured epithelial cells: effects of expression of exogenous N-ras oncogene.

We examined the functional consequences of cellular transformation of rat IAR-2 epithelial cells, by a mutant N-ras oncogene, on the dynamics of active lamellae, structures that play an important role in cell motility, adhesion, and surface-receptor capping. Lamellar activity was assessed by measuring the rate of outer-edge pseudopodial activity and by analyzing the motility of Con A-coated beads placed on lamellar surfaces with optical tweezers. Although transformation dramatically affected the shape and size of active cellular lamellae, there was little detectable effect on either pseudopodial activity or bead movement. To investigate the potential relationship between functional lamellar activity and the microtubule cytoskeleton, lamellar activity was examined in nontransformed and transformed cells treated with the microtubule-disrupting drug nocodazole. In the absence of microtubules, transformed cells were less polarized and possessed decreased rates of pseudopodial and bead motility. On the basis of these observations, it is suggested that ras-induced transformation of epithelial cells consists of two cytoskeletal modifications: overall diminished actin cytoskeletal dynamics in lamellae and reorganization of the microtubule cytoskeleton that directs pseudopodial activity to smaller polarized lamellae.

Animals↗
PubMed1994

Role of the microtubular system in morphological organization of normal and oncogene-transfected epithelial cells.

To understand better the role of the microtubular system in the development and maintenance of morphological organization of nonpolarized and polarized cells of the same origin we examined the effects of two microtubule-specific drugs, colcemid and taxol, on discoid cultured epithelial rat cells of the IAR-2 line and on polarized cells obtained from this line by transfection of mutated N-ras oncogene; morphometric, immunomorphologic, and videomicroscopic methods were used. Depolymerization of microtubules by colcemid did not cause major changes in the discoid shape of IAR cells but altered organization of actin cortex; in particular, it led to disappearance of circumferential bundle of actin microfilaments. Taxol reorganized the normal network of microtubules radiating from the perinuclear centers into numerous arrays of short microtubules not associated with any centers. Taxol-treated cells had wider circumferential bundles of microfilaments than control cells and morphometric analysis showed that their contours were closer to geometric circle than those of control or of colcemid-treated cells. These data show that function of the microtubular system is essential for maintenance of the characteristic morphological organization of discoid cells; we propose to name this function "contra-polarization." Contra-polarization is not prevented and is even promoted by taxol; this result suggests that a decentralized system of microtubules is sufficient for this function. In contrast, maintenance of polarized morphology of IAR-2 cells transfected by the N-ras oncogene is inhibited not only by colcemid but also by taxol and thus requires the presence of a normal centralized microtubular system.

Animals↗
PubMed1992

Reversible structural alterations of undifferentiated and differentiated human neuroblastoma cells induced by phorbol ester.

Morphological alterations in the structure of undifferentiated and morphologically differentiated human neuroblastoma cells induced by phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, were examined by video microscopy and immunomorphology. In undifferentiated cells, PMA induced the formation of motile actin-rich lamellas and of stable cylindrical processes rich in microtubules. Formation of stable processes resulted either from the collapse of lamellas or the movement of the cell body away from the base of a process. In differentiated cells, PMA induced the rapid extension of small lamellas and subsequent formation of short-lived elongated processes from the lateral edges of neurites. Additionally, growth cones exhibited enhanced modulation in shape after PMA treatment. These reversible reorganizations were similar to the actinoplast-tubuloplast transformations exhibited by PMA-treated fibroblasts. We suggest that actinoplast-tubuloplast reorganizations play essential roles in morphogenesis where stable cytoplasmic extensions are induced by external stimuli. In particular, PMA-induced reorganizations of neural cells in culture may be a model for morphological modulations that occur in nerve tissue.

Actins↗
PubMed1991

Post-translational modification of microtubules is a component of synergic alterations of cytoskeleton leading to formation of cytoplasmic processes in fibroblasts.

The protein kinase C activator phorbol 12-myristate 13-acetate (PMA) induces rapid and reversible shape changes in cultured fibroblasts: extension of motile lamellas is followed by transformation of these lamellas into nonmotile stalklike processes. This "lamella-to-stalk" transformation was found to be associated with the formation of microtubules enriched in detyrosinated alpha-tubulin. This change was local: microtubules in motile lamellas at the distal ends of the processes and in the cell bodies were not enriched in detyrosinated alpha-tubulin. Detyrosinated microtubules in the processes were more resistant to Colcemid treatment than other microtubules of PMA-treated and control cells. The effects of PMA were reversible and could be abolished by sphingosine, a specific inhibitor of protein kinase C. Besides modification of microtubules, lamella-to-stalk transformation is associated with the ingrowth of intermediate filaments into the extensions. Earlier it was found that this transformation is also associated with the profound reorganization of the system of actin microfilaments. Thus, all three cytoskeletal systems are altered simultaneously during PMA-induced formation of processes. Similar "cytoskeletal synergies" may play essential roles in many morphogenetic processes--e.g., in the growth of neurites.

Animals↗
PubMed1990

Cytoskeletal reorganizations responsible for the phorbol ester-induced formation of cytoplasmic processes: possible involvement of intermediate filaments.

The tumor promoter phorbol 12-myristate 13-acetate (PMA) induces characteristic reversible changes of cell shape in certain fibroblastic lines: motile lamellas are transformed into noncontractile narrow processes; simultaneously, the actin microfilament network of lamellas is locally disorganized. This reaction to PMA may be regarded as a prototype of reorganizations involving formation of stable cytoplasmic processes. Specific drugs, Taxol and Colcemid, were used to study the role of microtubules and vimentin-containing intermediate filaments (IF) in the development of PMA-induced reorganizations. PMA readily induced formation of noncontractile processes in Taxol-treated fibroblasts; these cells had a profoundly altered microtubular system but noncollapsed IF. A short (1 hr) exposure to PMA induced formation of processes in control cells but not in the Colcemid-treated cells, which had depolymerized microtubules and IF that collapsed around the nucleus. Longer (3-4 hr) exposure of the Colcemid-treated cells to PMA induced partial reversal of the IF collapse; those parts of the peripheral lamellas that contained IF were transformed into narrow noncontractile processes. It is suggested that the local interaction of IF with the actin system is an essential step in the formation of processes from lamellas. The microtubular system controls distribution of IF in the cytoplasm and thus plays an indirect role in the reorganization of the actin cortex.

Alkaloids↗
PubMed1988

Membrane-induced segregation of motile and stable domains in cytoplasm: possible role in morphological differentiation of tissue cells.

Many morphological differentiations and modulations involve alterations of degree of segregation in two types of cytoplasmic domains: lamellar-pseudopod-forming motile structures, actinoplasts, and non-motile cylindrical processes, tubuloplasts. Pronounced reversible segregation of actinoplasts and tubuloplasts can be induced in certain fibroblastic cells by phorbol ester. Analysis of this model reorganization suggests that formation of tubuloplasts from actinoplasts may involve local collapse of actin network and microtubule-directed translocation of actin material from collapsing zone; these cytoskeletal alterations may be controlled by membrane-activated regulatory enzymes.

Actins↗
PubMed1988

Microtubule-dependent effect of phorbol ester on the contractility of cytoskeleton of cultured fibroblasts.

The effect of the tumor promoter phorbol 12-myristate 13-acetate (PMA) upon the contractility of permeabilized cell models (cytoskeletons) of mouse fibroblasts was examined. Contraction was induced by incubation of cell models in a solution containing ATP and was assessed quantitatively by measuring alterations of the area of cell model projection on the substrate. Immunofluorescence microscopy was used to assess alterations of cytoskeleton morphology in the course of permeabilization and contraction. It was found that contractility of cell models from PMA-treated fibroblasts was considerably diminished as compared with the models from control fibroblasts. ATP induced only local contraction of certain zones of actin cortex in models from PMA-treated fibroblasts; it did not induce general contraction, characteristic of control models. Normal high contractility was characteristic of the models from the cells preincubated with PMA in combination with Colcemid. PMA is a specific activator of protein kinase C, one of the key enzymes of the membrane signal-transduction pathway. It is suggested that protein kinase C regulates contractility of actin cortex and that the pathway of this regulation has a microtubule-dependent stage blocked by Colcemid.

Adenosine Triphosphate↗
PubMed1987

Migration of nuclei and perikaryal cytoplasm along the cytoplasmic processes of differentiated neuroblastoma cells.

Time-lapse microcinematography of the cultures of mouse neuroblastoma C-1300 revealed several cases of directional translocation of nucleus-containing cell body along the elongated cytoplasmic processes of differentiated neuron-like cells. This unusual type of intracellular movement observed in vitro is similar to perikaryal translocation previously described in certain types of developing brain neurons in vivo. Translocation of nuclei was often accompanied by local contractions of surrounding cytoplasm. Repeated contractions and relaxations of cytoplasm were also observed in the same cultures in the course of transformation of undifferentiated well-spread neuroblast-like into differentiated neuron-like cells. Possible mechanisms of perikaryal translocations are discussed.

Animals↗
PubMed1987

Special type of morphological reorganization induced by phorbol ester: reversible partition of cell into motile and stable domains.

The phorbol ester phorbol 12-myristate 13-acetate (PMA) induced reversible alteration of the shape of fibroblastic cells of certain transformed lines--namely, partition of the cells into two types of domains: motile body actively extending large lamellas and stable narrow cytoplasmic processes. Dynamic observations have shown that stable processes are formed from partially retracted lamellas and from contracted tail parts of cell bodies. Immunofluorescence microscopy and electron microscopy of platinum replicas of cytoskeleton have shown that PMA-induced narrow processes are rich in microtubules and intermediate filaments but relatively poor in actin microfilaments; in contrast, lamellas and cell bodies contained numerous microfilaments. Colcemid-induced depolymerization of microtubules led to contraction of PMA-induced processes; cytochalasin B prevented this contraction. It is suggested that PMA-induced separation of cell into motile and stable parts is due to directional movement of actin structures along the microtubular framework. Similar movements may play an important role in various normal morphogenetic processes.

Animals↗
PubMed1985

Polarization of cytoplasmic fragments microsurgically detached from mouse fibroblasts.

We have studied the polarity of cytoplasm organization in tiny fragments of mouse embryo fibroblasts, produced by the microsurgical separation of long processes of cytochalasin-treated cells. In the cytochalasin-free medium fragments respread and developed small lamellas at one or both of their ends. Granules, visible at phase-contrast optics, were always collected in the central part of the fragment. Lamellas of the fragment, as well as lamellar cytoplasm of parent cells, were able to clear surface receptors patched by concanavalin A and an antibody to concanavalin A. Immunofluorescence microscopy showed that the fragments always contained actin microfilament bundles parallel to the long axis of the fragment, but microtubules were present not more than for 6 hrs after detachment of the fragments from the cell bodies. Fragments detached from the cells treated with colcemid and cytochalasin simultaneously and transferred into the drug-free medium never had any microtubules. In spite of that, their behaviour was similar to the behaviour of the fragments that were produced from the control cells treated only with cytochalasin. These results show that the small fragments of mouse embryo fibroblasts are able to maintain the polar organization of cytoplasm and the microtubules are not responsible for this organization.

Animals↗
PubMed1985

Effect of microtubule-destroying drugs on the spreading and shape of cultured epithelial cells.

The role of microtubules in the spreading of cells from the liver-derived IAR2 rat cell line was studied. Cells in the control medium seeded on a flat isotropic glass surface rapidly spread to form discoid shapes. Spreading in colcemid-containing medium was disorganized and delayed; partial reversal of spreading was observed. Nevertheless, even in the presence of colcemid the cells finally spread to discoid flattened shapes. IAR2 cells in medium without colcemid spread not to discoid but to elongated shapes under three different sets of conditions: (1) when the cells were forced to spread on narrow strips of adhesive glass surface between two non-adhesive lipid films; (2) when the cells spread on the poorly adhesive surface of poly(HEMA)-covered glass; (3) when the cells spread on the usual glass surfaces in medium containing cytochalasin D. Addition of colcemid to the media reversed the polarized spreading under the first two conditions; colcemid did not reverse the formation of the elongated cell shape acquired by the cells spreading in cytochalasin-containing medium. Effects of microtubule-destroying drugs on the spreading of epithelial and fibroblast cells are compared and discussed. It is suggested that microtubules are essential for the stabilization of the spread state of those attached cytoplasmic processes and lamellae that do not have numerous and stable-cell substratum contacts, e.g. the processes formed at the early stages of spreading or the elongated processes of polarized cells. Possibly, microtubules stabilize the non-contracted state of the actin cytoskeleton in these processes.

Animals↗
PubMed1984

Origin of signals conveyed by the ventral spino-cerebellar tract and spino-reticulo-cerebellar pathway.

The "fictitious" scratch reflex was evoked in decerebrate curarized cats by pinna stimulation. Activity of neurons of the ventral spino-cerebellar tract ( VSCT ) from the L4 and L5 segments of the spinal cord as well as of neurons of the spino-reticulo-cerebellar pathway ( SRCP ) from the lateral reticular nucleus of the medulla oblongata was recorded. Cooling and destruction of different parts of the lumbo-sacral enlargement of the spinal cord were performed. Cooling of the L5 or L6 segment abolished the rhythmic activity in the greater part of the spinal hindlimb centre but did not affect the generation of rhythmic oscillations in the remaining (rostral) segments of the lumbo-sacral enlargement. Under these conditions, neither the rhythmic activity of VSCT neurons located rostral to the thermode nor that of SRCP neurons changed. A normal rhythmic activity of SRCP neurons also persisted after destruction of grey matter in the L3 and L4 segments. It can be concluded that activity of these neurons is independent of whichever part of the enlargement generates rhythmic oscillations. From these observations a hypothesis is advanced that the main content of signals conveyed by the VSCT and SRCP to the cerebellum is the information regarding activity of the generator of rhythmic oscillations that is located in the L3-L5 spinal segments.

Animals↗
PubMed1984

Multinucleation-induced improvement of the spreading of transformed cells on the substratum.

Multinucleation of various cultured cells was produced by polyethylene glycol-induced fusion or by cytochalasin-induced block of mitosis. It was found that multinucleation induced by both methods considerably improved deficient spreading of all the tested transformed fibroblastic lines; average substratum area occupied by one cell and divided per number of nuclei was 2.0-2.5 times larger for multinucleated cells than for mononucleated ones. Improved spreading was accompanied by increased area of lamellar cytoplasm, increased number of focal contacts, and, in certain lines, by the appearance of actin bundles; numerous microtubules and intermediate filaments radiated from perinuclear zones into the lamellas of multinucleated cells. The number of cell-associated fibronectin fibrils was not increased by multinucleation. Cycloheximide did not prevent the improvement of spreading, suggesting that this effect was not due to any alterations of protein synthesis. Colcemid considerably decreased the effect of multinucleation but did not abolish it completely. It is suggested that increase of spreading is due to multinucleation-associated alterations of quantitative interrelationships between various cell components. One of these alterations is probably increased density of microtubules per unit length of outer cell edge.

Animals↗
PubMed1982

Effects of small doses of cytochalasins on fibroblasts: preferential changes of active edges and focal contacts.

The effects of low doses of cytochalasin B (2 micrograms/ml) and cytochalasin D (0.2 microgram/ml) on the spreading of normal mouse fibroblasts in culture were investigated to find out which components of cell-substrate interactions are most sensitive to alterations of the state of actin cytoskeleton. Cytochalasin B disorganized the cortical layer of actin microfilaments and caused partial or complete disappearance of microfilament bundles; focal contacts with the substrate seen by interference-reflection microscopy also disappeared. Diffuse close contacts were apparently insensitive to cytochalasin B. Low doses of cytochalasin B did not inhibit the outgrowth and maintenance of lamellas at the cell periphery. However, in contrast to controls, these lamellas had no distal zones with convex outer edges and ruffles at the upper surfaces. The disappearance of these ruffling active edges was accompanied by loss of the ability to clear the surface of the lamellas from the concanavalin A receptors crosslinked by the corresponding ligand. The effects of cytochalasin D were similar to those of cytochalasin B. Thus, ruffling active edges and focal contacts can be regarded as specialized parts of lamellas with increased sensitivity to cytochalasins; the presence of ruffling active edges is essential for the initiation of centripetal movement of the patches of crosslinked surface receptors.

Actins↗
PubMed1982

Possible common mechanisms of morphological and growth-related alterations accompanying neoplastic transformation.

Two main groups of phenotypic alterations usually accompany neoplastic transformations of cultured fibroblastic and epithelial cells: alterations of growth control and decreased formation of cell-substrate and cell-cell contacts. We suggest that both types of alterations are due to change of cell response to clustered membrane receptors. Clustering of certain receptors by corresponding ligands possibly induces a dual set of cellular reactions: (i) activation of cell proliferation preceded by an ordered sequence of prereplicative changes and (ii) attachment-clearing reactions that are associated with anchoring of ligand-receptor complexes by cytoskeletal components and may eventually lead to internalization of these complexes. The first stages of these two subsets of dual reactions are probably identical, but later the two chains of reactions are separated; completion of clearing reaction may stop further progress of the activation of proliferation. Attachment of the cell membrane to another surface can be regarded as a variant of unfinished clearing reaction. Transformed cells may be characterized by alterations of the dual reactions; the primary change may affect either the attachment-clearing branch or the common early stages preceding the separation of branches. These alterations may lead to facilitated activation of proliferation and also to primary or secondary decrease of the ability to perform attachment-clearing reactions in response to external ligands.

Animals↗
PubMed1980

Spreading of fibroblasts in medium containing cytochalasin B: formation of lamellar cytoplasm as a combination of several functional different processes.

Normal cultured mouse fibroblasts spreading on solid substrate extend and attach numerous pseudopods; lamellar cytoplasm is eventually formed from the attached pseudopods. Fibroblasts spreading in the presence of cytochasin B (CB) from de novo a system of arbor-like branched processes rather than lamellar cytoplasm. The growing and fully formed arbor-like processes, in contrast to normal lamellar cytoplasm, have low contractility and are unable to clear patched concanavalin A receptors from their surfaces; their attachement sites are not associated with microfilament bundles. The cells spreading in medium containing CB and Colcemid do not form well-organized branched structures but extend and attach numerous unstable pseudopods. It is suggested that normal formation of lamellar cytoplasm can be regarded as a combination of several functionally different processes: (a) of rudimentary pseudopodial reactions resistant to CB and Colcemid; (b) of CB-sensitive lamellization; and (c) of Colcemid-sensitive stabilization.

Animals↗