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M Wallin

Publications and source records attributed to M Wallin.

At least 37 records · Page 2Linked to original sources
PubMed1996

Localization of kinesin and cytoplasmic dynein in cultured melanophores from Atlantic cod, Gadus morhua.

In this study we have analyzed pigment translocation in cultured melanophores from the cold-tempered Atlantic cod, Gadus morhua. The transport process was found to be cold-adapted, as it proceeded at low temperatures. Both the typical morphology of the melanophores with long cytoplasmic processes, and the ability to translocate pigment granules, were found to be highly dependent on microtubules. Microtubules in melanophores were relatively stable to vinblastine treatment compared to microtubules in other skin cells. Extensive posttranslational modifications of tubulin were found. Detyrosinated and polyglutamylated microtubules were frequent, while acetylated microtubules only comprised a subpopulation or domains of microtubules. Both cod kinesin and dynein were distributed in a punctate pattern throughout the melanophores in close proximity to microtubules. The motors accumulated together with pigment granules during aggregation and were dispersed during translocation of pigment granules to the periphery. Individual melanosomes were occasionally found to rapidly change direction during translocation. Our data raise the interesting possibility that both kinesin and dynein are bound to pigment granules. This is of functional significance, since pigment granules are transported back and forth in the melanophores, and may be activated differently during aggregation and dispersion to generate translocation.

Animals↗
PubMed1996

Detyrosination of tubulin is not correlated to cold-adaptation of microtubules in cultured cells from the Atlantic cod (Gadus morhua).

Isolated cod brain microtubules from the cold-adapted Atlantic cod (Gadus morhua) have previously been shown to be highly detyrosinated, a post-translational modification of tubulin usually found in stable subsets of microtubules. In this study we found this was not restricted only to isolated brain microtubules. Microtubules in primary cultures of brain and skin cells were composed of both tyrosinated (Tyr)- and detyrosinated (Glu)-tubulin seen by immunocytochemistry. Immunoelectron microscopy of isolated microtubules showed that individual microtubules were composed of a mixture of Tyr- and Glu-tubulin. Leukocytes with extending lamellopodia contained only microtubules stained with the antibody against Tyr-tubulin, and isolated heart tubulin lacked both Tyr- and Glu-tubulin, suggesting that a relative high level of detyrosination is a characteristic of most, but not all, cod microtubules. Brain cell microtubules were more resistant to mitotic inhibitors than skin cell microtubules, but this was not correlated to a difference in detyrosination. Brain and skin cell microtubules were only partially disassembled when incubated at 0 degrees C. Upon reassembly of microtubules at 12 degrees C, microtubules were still made of mixtures of Tyr- and Glu-tubulin, indicating that detyrosination of assembled microtubules is rapid and/or that in cod cells in contrast to mammalian cells, Glu-tubulin can reassemble to microtubules. Our data show that most cod microtubules are highly detyrosinated, but this is not the cause of their cold adaptation or drug stability.

Animals↗
PubMed1995

Glucocorticoid receptor inhibits microtubule assembly in vitro.

The effect of glucocorticoid hormones, purified glucocorticoid receptor (GR) and purified heat shock protein M(r) 90,000 (hsp90) on microtubule (MT) assembly in vitro was tested by a spectrophotometric MT assembly assay and electron microscopy. GR significantly prolonged the nucleation phase, slowed down the assembly rate and reduced the maximal amplitude of MT assembly compared with control. The effects were partially reversed by the addition of glucocorticoid hormone. GR associated with MTs. These results indicate that GR affects MT assembly in vitro, which may be a functional correlate to the structural association of GR with MTs. This implies that factors affecting GR may affect MT assembly in vivo.

Animals↗
PubMed1995

Comparative study of the colchicine binding site and the assembly of fish and mammalian microtubule proteins.

Isolated microtubules from cod (Gadus morhua) are apparently more stable to colchicine than bovine microtubules. In order to further characterize this difference, the effect of the colchicine analogue 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6-cyclo heptatrien-1-one (MTC) was studied on assembly, as measured by turbidity and sedimentation analysis, and on polymer morphology. MTC has the advantage to bind fast and reversible to the colchicine binding site of tubulin even at low temperatures. It was found to bind to one site in cod brain tubulin, with affinity (6.5 +/- 1.5) x 10(5)M-1 at both low or high temperature, similarly to bovine brain tubulin. However, the effect of the binding differed. At substoichiometric concentrations of MTC bovine brain microtubule assembly was almost completely inhibited, while less effect was seen on the mass of polymerized cod microtubule proteins. A preformed bovine tubulin-colchicine complex inhibited the assembly of both cod and bovine microtubules at substoichiometric concentrations, but the effect on the assembly of cod microtubules was less. At higher concentrations (5 x 10(-5) to 1 x 10(-3) M), MTC induced a large amount of cold-stable spirals of cod proteins, whereas abnormal polymers without any defined structure were formed from bovine proteins. Spirals of cod microtubule proteins were only formed in the presence of microtubule associated proteins (MAPs), indicating that the morphological effect of MTC can be modulated by MAPs. The effects of colchicine and MTC differed. At 10(-5) M colchicine no spirals were formed, while at 10(-4) M and 10(-3) M, a mixture of spirals and aggregates was found. The morphology of the spirals differed both from vinblastine spirals and from the spirals previously found when cod microtubule proteins polymerize in the presence of high Ca2+ concentrations. The present data show that even if the colchicine binding site is conserved between many different species, the bindings have different effects which seem to depend on intrinsic properties of the different tubulins.

Animals↗
PubMed1995

Distribution of acetylated tubulin in cultured cells and tissues from the Atlantic cod (Gadus morhua). Role of acetylation in cold adaptation and drug stability.

The Atlantic cod (Gadus morhua) is a poikilothermic animal living at temperatures between 2-15 degrees C. Isolated cod brain tubulin is, in contrast to mammalian brain tubulin, posttranslationally modified by acetylation to a high extent. To investigate the role of acetylation in cold adaptation, microtubules were isolated by a taxol-dependent procedure from different organs of the cod, and cells from different tissues were cultured. All cells from skin and brain were able to grow between 4 degrees C and room temperature. Microtubules in the cultured cells were sometimes severed near the periphery of the cells. Microtubules in brain cells were in general more stable to vinblastine and colchicine, when compared to skin cells. Acetylated microtubules were found only in brain cells, in peripheral nerves on scales and in nerves of the intestinal tract and in microtubules isolated from neuronal tissue. Our results show that acetylated microtubules are found both in the central and peripheral nervous system, but that there is no correlation between acetylation and cold-adaptation.

Acetylation↗
PubMed1995

Cold-stable and cold-adapted microtubules.

Most mammalian microtubules disassemble at low temperature, but some are cold stable. This probably has little to do with a need for cold-stable microtubules, but reflects that certain populations of microtubules must be stabilized for specific functions. There are several routes by which to achieve cold stability. Factors that interact with microtubules, such as microtubule-associated proteins, STOPs (stable tubule only polypeptides), histones, and possibly capping factors, are involved. Specific tubulin isotypes and posttranslational modifications might also be of importance. More permanent stable microtubules can be achieved by bundling factors, associations to membranes, as well as by assembly of microtubule doublets and triplets. This is, however, not the explanation for cold adaptation of microtubules from poikilothermic animals, that is, animals that must have all their microtubules adapted to low temperatures. All evidence so far suggests that cold adaptation is intrinsic to the tubulins, but it is unknown whether it depends on different amino acid sequences or posttranslational modifications.

Adaptation, Physiological↗
PubMed1994

Different stability of posttranslationally modified brain microtubules isolated from cold-temperate fish.

Microtubule proteins were isolated by a temperature-dependent assembly-disassembly method from brain tissue of for cold-temperature fish; one fresh water fish (Oncorhynchus mykiss), and three marine fish (Labrus berggylta, Zoarces viviparus and Gadus morhua). The alpha-tubulins from all four fish species were acetylated. The alpha-tubulins from the marine fish were composed of a mixture of tyrosinated and detyrosinated tubulin, while the fresh water fish tubulin only reacted with an antibody against detyrosinated tubulin. The isolated microtubules had a similar MAP composition. A 400 kD protein and a MAP2-like protein were found, but MAP1 was missing. All microtubules disassembled upon cooling to 0 degrees C. In spite of these common characteristics, the assembly of microtubules from Labrus berggylta was inhibited by colchicine and calcium, in contrast to the assembly of microtubules from Oncorhynchus mykiss and Zoarces viviparus. For the latter, colchicine was not completely inhibitory even at a concentration as high as 1 mM, and calcium induced the formation of both loosely and densely coiled ribbons. The effects of calcium and colchicine on microtubules from Oncorhynchus mykiss and Zoarces viviparus were modulated by either fish or cow MAPs, indicating that the effects are due to intrinsic properties of the fish tubulins and not the MAPs. In view of these findings, our results suggest that there is no correlation between colchicine sensitivity, inability of calcium to inhibit microtubule assembly, and acetylation and detyrosination.

Acetylation↗
PubMed1994

Differences in the effect of Ca2+ on isolated microtubules from cod and cow brain.

Isolated microtubules from cod and cow brains were compared with respect to their response to calcium ions. The effect of Ca2+ on cod microtubules was found to be temperature dependent. In contrast to cow microtubules, cod microtubules assembled at 18 degrees C. At this temperature the assembly was inhibited by Ca2+ concentrations of 2 mM and higher. This was also found for cow microtubules at 37 degrees C. However, at 30 degrees C there was no effect of 2 mM Ca2+ of the amount of assembly or disassembly of cod microtubules consisting of only tubulin or of tubulin and microtubule-associated proteins (MAPs). The morphology was affected though, since some coiled ribbons formed from tubulin and MAPs. The calcium-binding calmodulin did not alter the effect of calcium on cod microtubules markedly. At higher Ca2+ concentrations (> 4 mM), coiled ribbons were formed from cod tubulin and MAPs, but mainly amorphous aggregates and very few coiled ribbons were formed from cod tubulin alone, indicating that the Ca2+ effect is modulated by cod MAPs. The modulatory effect of cod MAPs was however not species specific, since both cod and cow MAPs had the same effect on cod microtubules, in spite of a different protein composition. A MAP-dependent effect of Ca2+ was also found for cow microtubule proteins. The assembly of pure cow tubulin, as well as that of cow tubulin and MAPs, was inhibited by 2 mM Ca2+. In the presence of 10 and 20 mM Ca2+, pure cow tubulin formed amorphous aggregates, rings, and even paracrystals, while the assembly of cow tubulin and MAPs was inhibited. Our results suggest therefore that the effect of Ca2+ can be moderated by MAPs, but depends on intrinsic properties of the different tubulins.

Animals↗
PubMed1994

Dynamic instability of microtubules from cold-living fishes.

The dynamic instability of microtubules free of microtubule-associated proteins from two genera of cold-living fishes was measured, by means of video-enhanced differential-interference-contrast microscopy, at temperatures near those of their habitats. Brain microtubules were isolated from the boreal Atlantic cod (Gadus morhua; habitat temperature approximately 2-15 degrees C) and from two austral Antarctic rockcods (Notothenia gibberifrons and N. coriiceps neglecta; habitat temperature approximately -1.8 to + 2 degrees C). Critical concentrations for polymerization of the fish tubulins were in the neighborhood of 1 mg/ml, consistent with high interdimer affinities. Rates of elongation and frequencies of growth-to-shortening transitions ("catastrophes") for fish microtubules were significantly smaller than those for mammalian microtubules. Slow dynamics is therefore an intrinsic property of these fish tubulins, presumably reflecting their adaptation to low temperatures. Two-dimensional electrophoresis showed striking differences between the isoform compositions of the cod and the rockcod tubulins, which suggests that the cold-adapted microtubule phenotypes of northern and southern fishes may have arisen independently.

Animals↗
PubMed1994

Do lowered factor VII levels at extremely high endogenous oestradiol levels protect against thrombin formation?

The purpose of the study was to find a model to study the effect of endogenous oestradiol on haemostasis. Hormones and haemostatic variables were therefore measured before and after in vitro fertilization and egg replacement therapy in 14 women with tubal infertility. The differences between the levels of the haemostatic variables at minimum and maximum levels of oestradiol and progesterone were evaluated. A significant increase in the plasma levels of coagulation factor VIII, von Willebrand factor and fibrinogen was found between samples drawn before stimulation and at the highest oestradiol levels (P < 0.002, P < 0.002 and P < 0.015, respectively). However, coagulation factor VII activity and antigen decreased significantly (P < 0.001 and P = 0.009, respectively). The levels of the coagulation inhibitors protein C and antithrombin decreased (P < 0.003 and P = 0.008, respectively), while that of free protein S increased (P = 0.039). No significant changes were observed in the fibrinolytic variables or in those reflecting thrombin activity (prothrombin fragment 1 + 2, thrombin-antithrombin complexes, soluble fibrin and D-dimers). In conclusion, the increase in the levels of factor VIII, von Willebrand factor, fibrinogen and the decrease in the levels of antithrombin and protein C may, if coagulation is triggered, contribute to a hypercoagulable state. The depressed factor VII levels and the tendency to elevated free protein S levels may constitute a protective mechanism that modulates the coagulation system when levels of oestradiol become extremely high.

Blood Coagulation Disorders↗