Biomedical subjects

M Bastmeyer

Publications and source records attributed to M Bastmeyer.

34 records · Page 2Linked to original sources

PubMed1994

Similarities and differences between fish oligodendrocytes and Schwann cells in vitro.

In light of the striking differences between oligodendrocytes of the optic nerve/tract of adult goldfish and their mammalian counterparts, a further characterization of goldfish oligodendrocytes was performed. A comparison with Schwann cells was included because fish optic nerve/tract-derived oligodendrocytes bear remarkable similarities to this type of glial cell. Fish optic nerve/tract-derived oligodendrocytes that had differentiated into 04 and 6D2-positive cells and thus expressed early myelin marker molecules were found to incorporate BrdU and to divide in vitro over a period of weeks. For the induction of more advanced markers of myelinogenesis such as the CNS-specific myelin protein 36K, oligodendrocytes required extensive contact with axons. Other agents, such as fetal calf or carp serum, substrate components, or forscolin failed, however, to induce 36K expression. 04/6D2-positive oligodendrocytes could be distinguished from fish 6D2-positive Schwann cells derived from cranial nerves by their antigenic phenotype: Schwann cells but not oligodendrocytes exhibited the low affinity NGF receptor. While both cell types carry the cell adhesion molecules NCAM, E 587 antigen, and the L2/HNK-1 epitope, only Schwann cells possess a further adhesion molecule, Neurolin.

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PubMed1993

Fish optic nerve oligodendrocytes support axonal regeneration of fish and mammalian retinal ganglion cells.

Segments from adult fish and rat retinae were explanted on myelin-marker expressing oligodendrocytes derived from the regenerating goldfish optic nerve. Fish axons grew in high density and even rat retinal axons regenerated to considerable length on the surface of the fish oligodendrocytes, suggesting that this type of fish glia has axon-growth promoting surface components that exert their influence across species boundaries. One interesting surface component of the fish oligodendrocytes as demonstrated here is the E 587 antigen, which is related to the L1 family of cell adhesion molecules. In long term cocultures of oligodendrocytes and retinal axons, the fish glial cells were found to enwrap rat axons. This suggests that the oligodendrocytes of the regenerating goldfish optic nerve/tract may, despite striking differences, represent the equivalent to mammalian optic nerve oligodendrocytes.

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PubMed1993

Behavior of fish retinal growth cones encountering chick caudal tectal membranes: a time-lapse study on growth cone collapse.

In a cross species in vitro assay, growth cones from fish temporal retina elongating on laminin lanes were observed with time-lapse videomicroscopy as they encountered lanes and territories that carried membrane fragments from the chick caudal tectum. Caudal tectal membranes of adult fish and embryonic chick are known to possess a repellent guiding component for temporal retinal axons. The caudal membranes of chick exert a particularly strong influence on fish temporal axons. Contacts with chick caudal membranes by just a few filopodia and parts of the lamellipodia evoked a turning response away from the membrane lane of the entire growth cone. Contacts by filo- and lamellipodia over the entire circumference of the growth cone, however, caused invariably growth cone collapse and retraction. During growth cone turning and collapse and retraction, filopodia remained in contact with the tectal membrane fragments, suggesting strong membrane-filopodia adhesion simultaneous to growth cone repulsion by the repellent guiding component.

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PubMed1993

Expression of JUN, KROX, and CREB transcription factors in goldfish and rat retinal ganglion cells following optic nerve lesion is related to axonal sprouting.

Goldfish and rat optic nerves were cut and crushed, respectively, and the expression of the transcription factor proteins c-JUN, JUN B, JUN D, c-FOS, FOS B, KROX-24, and CREB was investigated in retinal ganglion cells (RGCs) by immunocytochemistry. Immunoreactivities (IRs) were followed up to 350 days in the goldfish and up to 22 days in the rat. In RGCs of untreated goldfish and rats, all JUN, FOS, and KROX proteins were absent whereas CREB was constitutively expressed. After optic nerve cut in goldfish, a JUN-like immunoreactivity (JUN-IR) appeared in a small number of RGCs of central retina after 24 h, reached a maximum within 5 days, declined after 30 days, and was on a half-maximal level after 50 days. Between 100 and 200 days, JUN-IR was only visible in a few RGCs and was completely absent after 350 days. Specific antibodies against c-JUN, JUN B, and JUN D gave no distinct immunoreactive signal. Thus, we could not determine which member of the JUN family contributed to the JUN-IR. The expression of CREB declined after 5 days. The number of CREB-labeled RGCs was reduced (not significant) and the intensity of labeling faded out. After 50 days, CREB-IR had returned to basal level. c-FOS, FOS B, and KROX-24 could not be detected in goldfish RGCs following optic nerve cut. After optic nerve crush in the rat, c-JUN, JUN D, and KROX-24 appeared in a substantial number of RGCs after 24 h, had a maximal expression after 5 days, and strongly declined after 8 days. c-JUN and KROX-24 were completely absent after 22 days whereas JUN D was still present in a few rat RGCs. The number of CREB-labeled RGCs decreased after 5 days and had declined by 50% after 22 days. Expression of JUN B, c-FOS, FOS B could not be detected in rat RGCs after optic nerve crush. Our data demonstrate that the decrease of CREB and the increase of JUN and KROX-24 transcription factors precedes and parallels both the alteration of de novo protein synthesis and the axonal sprouting, which are long lasting in goldfish and transient in rat.

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PubMed1992

Trying to understand axonal regeneration in the CNS of fish.

In contrast to the situation in mammals and birds, neurons in the central nervous system (CNS) of fish--such as the retinal ganglion cells--are capable of regenerating their axons and restoring vision. Special properties of the glial cells and the neurons of the fish visual pathway appear to contribute to the success of axonal regeneration. The fish oligodendrocytes lack the axon growth inhibiting molecules that interfere with axonal extension in mammals. Instead, fish optic nerve oligodendrocytes support--at least in vitro--axonal elongation of fish as well as that of rat retinal axons. Moreover, the fish retinal ganglion cells re-express upon injury a set of growth-associated cell surface molecules and equip the regenerating axons throughout their path and up into their target, the tectum opticum with these molecules. This may indicate that the injured fish ganglion cells reactivate the cellular machinery necessary for axonal regrowth and pathfinding. Furthermore, the target itself provides positional marker molecules even in adult fish. These marker molecules are required to guide the regenerating axons back to their retinotopic home territory within the tectum.

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PubMed1991

Two populations of glial cells from fish optic nerve/tract with distinct electrophysiological properties.

The electrophysiological properties of the two major glial cell types in cultures from the regenerating goldfish optic nerve/tract were studied with patch-clamp techniques. Spindle-shaped cells express myelin proteins. These oligodendrocyte-like cells possess outwardly rectifying currents, do not show glutamate activated currents and are rarely electrically coupled to neighboring cells. Cells of epitheloid morphology probably represent astrocytes. They are GFAP-positive and do not exhibit myelin proteins. These cells have glutamate activated currents, display a linear current to voltage relationship and are extensively electrically coupled thus displaying properties similar to mammalian astrocytes.

4-Aminopyridine↗

PubMed1991

Cytology of Lepidoptera. V. The microtubule cytoskeleton in eupyrene spermatocytes of Ephestia kuehniella (Pyralidae), Inachis io (Nymphalidae), and Orgyia antiqua (Lymantriidae).

The structural transformations of the microtubule cytoskeleton during eupyrene meiosis were characterized in the Mediterranean mealmoth, Ephestia kuehniella. Anti-tubulin immunofluorescence was used in two strains, L and Sbr, of this species. In addition, living primary eupyrene spermatocytes from strain L were observed with phase-contrast light microscopy, and metaphase I spermatocytes were studied with the electron microscope. The analysis did not reveal strain-specific differences in the behavior of the chromatin and the microtubule cytoskeleton in eupyrene meiosis. The general pattern of the two subsequent meiotic divisions in Ephestia kuehniella did not deviate from that observed during nuclear division in other species. A bipolar spindle developed during prophase, chromatin migrated towards the poles in anaphase, and spindle elongation occurred in telophase. Spindle development and behavior were qualitatively almost identical in primary and secondary spermatocytes. The assembly of the second meiotic spindle was initiated during the depolymerization of interzone microtubules of the first division. The relationship between spindle microtubules and centrosomes, however, showed some particularities in the moth. While microtubules radiated out from the spindle poles in late prophase, most microtubules ended about half-way between the equatorial plane and the spindle poles from late prometaphase to early anaphase. The termination of most microtubules distant from the poles was confirmed using electron microscopy in Ephestia kuehniella and using anti-tubulin immunofluorescence in two other Lepidoptera species, Inachis io and Orgyia antiqua. The microtubules were oriented parallel to the pole-to-pole axis of the spindle and were confined to an area laterally sheath by membrane layers. The spindle portions between the pole-proximal termini of the microtubules and the spindle poles contained irregularly shaped membrane elements. In late anaphase, microtubules reform between the migrating chromatin plates and the spindle poles. Concomitantly with the migration of the chromosomes towards the spindle poles, interzone microtubules are assembled. The origin of most of them appears to be at the pole-distal face of the chromatin masses.

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PubMed1991

Cytology of lepidoptera. VI. Immunolocalization of microtubules in detergent-extracted apyrene spermatocytes of Ephestia kuehniella Z.

Apyrene meiosis was studied in two wild-type strains, L and Sbr, of the Mediterranean mealmoth, Ephestia kuehniella, using anti-tubulin immunofluorescence. The observations were supplemented by phase-contrast light microscopy of living spermatocytes from strain L. The study revealed that nuclear envelope breakdown, centrosome separation, migration of chromatin elements towards the poles, and spindle elongation also occur in apyrene spermatocytes. However, a conventional metaphase plate is never formed, and chromatin segregation is irregular and delayed. Chromosome laggards are frequent. As a rule, apyrene spindles have a low microtubule content. The two strains, L and Sbr, differ regarding the chromatin behavior during meiosis. In strain L, the developing spindles contain numerous small chromatin clumps which segregate asynchronously. The resulting daughter cells possess about the same amount of chromatin. In contrast, large chromatin clumps exist in strain Sbr at the onset of spindle formation. The chromatin blocks transiently occupy an equatorial position and elongate subsequently parallel to the spindle axis. These elongated chromatin bodies often divide highly unequally. As a consequence, secondary spermatocytes in strain Sbr differ greatly in chromatin content. Subjective assessment shows that the size of the microtubular cytoskeleton is positively correlated with the chromatin content of the cell. Hence, it is hypothesized that the chromatin content determines spindle size. This possibly comes about the number of available kinetochores which are exposed and able to stabilize microtubules of centrosomal origin attached with the kinetochores. However, a direct bearing of chromatin on spindle size is similarly conceivable. Other Lepidoptera species examined so far are compatible with a 'type L' or a 'type Sbr' pattern of apyrene meiosis.

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PubMed1991

Growth of regenerating goldfish axons is inhibited by rat oligodendrocytes and CNS myelin but not but not by goldfish optic nerve tract oligodendrocytelike cells and fish CNS myelin.

Encounters of regenerating goldfish retinal axons with oligodendrocytes and CNS myelin of mammals and fish were monitored in in vitro assays. Upon contact with highly branched rat oligodendrocytes, goldfish axons collapsed or grew around but never crossed these cells. However, in the presence of the antibody IN-1 against the oligodendrocyte-associated growth-inhibitory proteins, axons did grow over highly branched oligodencrocytes. In contrast to the mammalian oligodendrocytes, goldfish optic nerve/tract-derived oligodendrocytelike cells allowed the growth of axons across their surface and even along their processes. The fish growth cones avoided entering the region of rat CNS myelin applied to polylysine/laminin-coated coverslips or failed to elongate on this substrate. They were, however, able to pass over CNS myelin of fish. When exposed to rat CNS myelin as the sole substrate, axonal outgrowth from fish retinal explants was inhibited almost entirely. However, outgrowth on fish CNS myelin was substantial, but many more axons extended on fish or rat brain membranes that were depleted of myelin. Thus, goldfish retinal axons are sensitive to the axon-growth-inhibiting cell-surface molecules of mammalian oligodendrocytes as well as CNS myelin. Fish optic nerve oligodendrocytelike cells and fish CNS myelin lack these inhibitory properties and are growth permissive. These in vitro experiments suggest that the success of axonal regeneration in the fish optic nerve is causally related to the presence of growth-permissive properties and to the absence of growth inhibitors on fish optic nerve/tract oligodendrocytelike cells.

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PubMed1990

The spatiotemporal distribution of N-CAM in the retinotectal pathway of adult goldfish detected by the monoclonal antibody D3.

The spatiotemporal distribution of neural cell adhesion molecule (N-CAM) in the retinotectal system of adult goldfish was assessed by immunofluorescence using the monoclonal antibody (Mab) D3 against chick N-CAM. In immunoblots with extracts of cell surface membranes of fish brains, Mab D3 recognized a prominent band at 170K and a weak band at 130K (K = 10(3) Mr). N-CAM immunofluorescence on cells was restricted to the marginal growth zones of the retina and the tectum and, in normal fish, to the youngest axons from the new ganglion cells of the peripheral retinal margin. In fish with previously transected optic nerves (ONS), Mab D3 staining was found transiently on all axons from the site of the cut into the retinorecipient layers of the tectum, but disappeared from these axons 450 days after ONS. Growing retinal axons in vitro exhibited N-CAM immunofluorescence throughout their entire extent, including their growth cones. Glial cells cultured from regenerating optic nerves were, however, unlabeled. These data are consistent with the idea that N-CAM is involved in adhesive interactions of growing axons. The temporally regulated expression of N-CAM on the new retinal axons may contribute to the creation of the age-related organization of the axons in the retinotectal pathway of fish.

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PubMed1989

Identification of astrocyte- and oligodendrocyte-like cells of goldfish optic nerves in culture.

Fish glial cells were obtained from cultivated segments of the optic nerve and raised in vitro. Two types of cells were identified as astrocyte- and oligodendrocyte-like glia by the monoclonal antibody Mab O1 (specific for oligodendrocytes) and the rabbit serum anti-goldfish glial fibrillary acidic protein (anti-G-GFAP). Cells of compact morphology were rare, and anti-G-GFAP positive and O1 negative. Multipolar cells in 5-day-old cultures were anti-G-GFAP but rarely O1 positive. In 5-week-old cultures, however, roughly 75% of the multipolar cells were double-labeled with both anti-G-GFAP and O1; 10% were single labeled with Mab O1 and 15% with anti-G-GFAP, respectively.

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PubMed1987

Characterization of Pales spermatocyte spindles, with reference to an MTOC-associated protein.

Pales ferruginea spermatocytes, because of their high proportion of dividing cells, offer many advantages in the study of the meiotic spindle. In the present study we have combined ultrastructural and immunofluorescence techniques with biochemical analysis of constituent spindle components. Using a centrosome-reactive serum, which recognizes an antigen in the centrosomal and midbody regions of dividing cells, we have identified an MTOC-associated polypeptide of 112,000 Mr with a pI of 5.6. The relative abundance of this protein in cells and isolated cytoskeletons indicates that it is present in other regions of the cell and is either more abundant, or is only recognized by the antibody in association with MTOCs.

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PubMed1986

The distribution of intermicrotubular bridges in meiotic spindles of the crane fly.

The distribution of intermicrotubular bridges in spindles of tipulid spermatocytes (Pales ferruginea, first meiotic division) was analyzed using serial sections of pre-selected cells. Bridges were found in all spindle regions, including kinetochore microtubules and free microtubules in the chromosome fiber. The dimensions of bridges were variable, ranging between 60 and 300 A in length and 40 and 190 A in thickness. Bridges seem to be randomly distributed. No accumulation in or absence from particular spindle regions was detected. Quantitative analysis revealed a linear, positive correlation between the number of microtubules and the number of microtubule pairs capable of forming bridges and, on the other hand, between microtubule pairs and intermicrotubular bridges. The possible composition and significance of bridges are discussed.

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PubMed1986

Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation?

Tipulid spermatocytes form normally functioning bipolar spindles after one of the centrosomes is experimentally dislocated from the nucleus in late diakinesis (Dietz, R., 1959, Z. Naturforsch., 14b:749-752; Dietz, R., 1963, Zool. Anz. Suppl., 23:131-138; Dietz, R., 1966, Heredity, 19:161-166). The possibility that dissociated pericentriolar material (PCM) is nevertheless responsible for the formation of the spindle in these cells cannot be ruled out based on live observation. In studying serial sections of complete cells and of lysed cells, it was found that centrosome-free spindle poles in the crane fly show neither pericentriolar-like material nor aster microtubules, whereas the displaced centrosomes appear complete, i.e., consist of a centriole pair, aster microtubules, and PCM. Exposure to a lysis buffer containing tubulin resulted in an increase of centrosomal asters due to aster microtubule polymerization. Aster-free spindle poles did not show any reaction, also indicating the absence of PCM at these poles. The results favor the hypothesis of chromosome-induced spindle pole formation at the onset of prometaphase and the dispensability of PCM in Pales.

Animals↗

PubMed1986

Immunostaining of spindle components in tipulid spermatocytes using a serum against pericentriolar material.

First meiotic division of tipulid (Pales ferruginea) spermatocytes was investigated by double immunostaining with anti-tubulin IgG and scleroderma 5051 serum against pericentriolar material (PCM). PCM-like material became visible in late diakinesis in centrosomal areas as well as in kinetochores. Anti-PCM fluorescence was most pronounced in metaphase and diminished again in anaphase. Displacement of one of the centrosomes from the nucleus at diakinesis in Pales spermatocytes leads to the formation of a bipolar, normally functioning spindle possessing one aster and centriole-free spindle pole (AFP). In about 80% of the AFPs observed there were no traces of anti-PCM staining detectable. This finding supports the assumption based on previous studies that polar PCM is not obligatory for spindle pole formation. The chromosomes seem to be able to induce the organization of a half-spindle. The strong anti-PCM fluorescence of the kinetochores observed here may be taken as further indication of tipulid chromosome autonomy regarding spindle formation.

Anaphase↗

PubMed1985

A model for chromosome movement based on lateral interaction of spindle microtubules.

Quantitative morphological studies of meiotic spindles in the crane fly Pales ferruginea (Fuge, 1980, 1984, 1985) were the basis for the development of a model explaining anaphase chromosome transport in higher eukaryotes. Two main features of chromosome fibres were important for the model: (1) the existence of microtubules oriented obliquely with respect to kinetochore microtubules, and (2) a higher degree of disorder in fibres exerting a pulling force. It is postulated that microtubules of the same polarity being inclined to each other at a certain angle are able to slide past each other by means of mechano-chemically active side-arms working in alternate succession. Sliding is suggested to lead to a displacement of microtubules and chromosomes in direction towards the poles. Furthermore, it is suggested that the chromosome fibre in anaphase becomes progressively disintegrated by fragmentation and disassembly of microtubules. Fragmentation may be induced by bending stress within the dynamic system.

Anaphase↗