Regulation of long-distance transport of mitochondria along microtubules

Mitochondria are cellular organelles of crucial importance, playing roles in cellular life and death. In certain cell types, such as neurons, mitochondria must travel long distances so as to meet metabolic demands of the cell. Mitochondrial movement is essentially microtubule (MT) based and is executed by two main motor proteins, Dynein and Kinesin. The organization of the cellular MT network and the identity of motors dictate mitochondrial transport. Tight coupling between MTs, motors, and the mitochondria is needed for the organelle precise localization. Two adaptor proteins are involved directly in mitochondria-motor coupling, namely Milton known also as TRAK, which is the motor adaptor, and Miro, which is the mitochondrial protein. Here, we discuss the active mitochondria transport process, as well as motor–mitochondria coupling in the context of MT organization in different cell types. We focus on mitochondrial trafficking in different cell types, specifically neurons, migrating cells, and polarized epithelial cells.     

A possible role of microtubules in the C cells secretory mechanism

Riassunto Le modalità di secrezione dei granuli contenenti Calcitonina da parte delle cellule C sono state studiate in tiroidi di cane in condizioni normali e in coltura organotipica con alto tenore di calcio. È stata notata la presenza di numerosi microtubuli alla periferia delle cellule e alcune immagini suggeriscono un attacco dei microtubuli ai granuli secretori. Viene prospettato che i microtubuli abbiano importanza nel meccanismo di secrezione delle cellule C e forse delle cellule della serie APUD in generale, e che questo possa essere del tipo «emiocitosi».     

The role of kinesin, dynein and microtubules in pancreatic secretion

The regulated secretion of pancreatic zymogens depends on a functional cytoskeleton and intracellular vesicle transport. To study the dynamics of tubulin and its motor proteins dynein and kinesin during secretion in pancreatic acinar cells, we infused rats with 0.1 μg/kg/h caerulein. Electron and fluorescence microscopy detected neither dynein nor kinesin at the apical secretory pole, nor on the surface of mature zymogen granules. After 30 min of secretagogue stimulation, kinesin and the Golgi marker protein 58 K were reallocated towards the apical plasma membrane and association of kinesin with tubulin was enhanced. Disruption of acinar cell microtubules had no effect on initial caerulein-induced amylase release but completely blocked secretion during a second stimulus. Our results suggest that mature zymogen granule exocytosis is independent of intact microtubules, kinesin and dynein. However, microtubule-dependent mechanisms seem to be important for the replenishment of secretory vesicles by redistribution of Golgi elements towards the apical cell pole. J. Schnekenburger and I.-A. Weber have contributed equally to this work.     

Staining of microtubules of the electrocyte ofElectrophorus electricus L. by alcian blue and lanthanum

Summary Microtubules were observed in the cytoplasm of theElectrophorus electricus L. when the tissue was fixed in the presence of alcian blue and lanthanum nitrate.Supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Conselho de Ensino para Graduados da UFRJ, and FINEP.     

Tubulin chaperone E binds microtubules and proteasomes and protects against misfolded protein stress

Mutation of tubulin chaperone E (TBCE) underlies hypoparathyroidism, retardation, and dysmorphism (HRD) syndrome with defective microtubule (MT) cytoskeleton. TBCE/yeast Pac2 comprises CAP-Gly, LRR (leucine-rich region), and UbL (ubiquitin-like) domains. TBCE folds α-tubulin and promotes α/β dimerization. We show that Pac2 functions in MT dynamics: the CAP-Gly domain binds α-tubulin and MTs, and functions in suppression of benomyl sensitivity of pac2Δ mutants. Pac2 binds proteasomes: the LRR binds Rpn1, and the UbL binds Rpn10; the latter interaction mediates Pac2 turnover. The UbL also binds the Skp1-Cdc53-F-box (SCF) ubiquitin ligase complex; these competing interactions for the UbL may impact on MT dynamics. pac2Δ mutants are sensitive to misfolded protein stress. This is suppressed by ectopic PAC2 with both the CAP-Gly and UbL domains being essential. We propose a novel role for Pac2 in the misfolded protein stress response based on its ability to interact with both the MT cytoskeleton and the proteasomes.     

Participation of microtubules and microfilaments in the transcellular biliary secretion of immunoglobulin A in primary cultures of rat hepatocytes

Summary The biliary secretion of immunoglobulin A (IgA) in primary hepatocyte cultures was investigated by means of immunofluorescence. The characteristic accumulation of IgA in visible bile canaliculi was found to be strongly inhibited by vinblastine and colchicine or by cytochalasin B, but its surface binding was not.The author thanks Prof. Dr D. Mecke for his support and encouragement. The excellent technical assistance of Mrs M. Behringer and Mrs A. Schneck is gratefully acknowledged. The author is grateful to the BIOTEST Serum Institut (Frankfurt) for the kind gift of human IgA. This work was supported by the Deutsche Forschungsgemeinschaft.     

Role of lysosomes, microtubules and microfilaments in the mechanism of the lactogenic action of prolactin in the rabbit mammary gland]

The lysosomotropic agents, ammonium chloride and chloroquine, added to the culture medium of pseudopregnant Rabbit mammary gland, did not inhibit the initiation of casein synthesis by prolactin. By contrast, they considerably reduced the down-regulation of prolacting receptors. Converse-y, colchicine totally blocked the lactogenic action of prolactin without altering the down-regulation of the receptor. Cytochalasin B inhibited only partly the lactogenic action of prolactin while it has no effect on the down-regulation of the receptor. These data suggest that the degradation of the prolactin-receptor complex in lysosome is not a compulsory step in the mechanism of prolactin action. The integrity of microtubules but not of microfilaments is required for prolactin to initiate casein synthesis. These elements of the cytoskeleton are not strictly involved in the down-regulation of the receptor.     

Taxuspine D,a new taxane diterpene fromTaxus cuspidata with potent inhibitory activity against Ca2+-induced depolymerization of microtubules

A new taxane diterpenoid, taxuspine D (1), possessing an enolacetate moiety, has been isolated from stems of the Japanese yewTaxus cuspidata Sieb. et Zucc., and the structure elucidated on the basis of spectroscopic data. Taxuspine D (1) markedly inhibited Ca²⁺-induced depolymerization of microtubules.     

Sur la présence de glycoprotéines dans les grains à microtubules des cellules sécrétrices paragoniales dedrosophila melanogaster meig

Summary Cytochemical observations of the paragonial microtubular granules during the first 2 weeks after emergence have shown the ultrastructural localization of glycoprotein in peripheral or central matrices by periodic acid-thiocarbohydrazide-silver proteinate method (PATAg). The microtubules do not appear to contain glycoprotein moiety. The functional significance of the components of paragonial secretion is discussed.     

Analysis of <Emphasis Type="Italic">Dictyostelium</Emphasis> TACC reveals differential interactions with CP224 and unusual dynamics of <Emphasis Type="Italic">Dictyostelium</Emphasis> microtubules

We have localized TACC to the microtubule-nucleating centrosomal corona and to microtubule plus ends. Using RNAi we proved that Dictyostelium TACC promotes microtubule growth during interphase and mitosis. For the first time we show in vivo that both TACC and XMAP215 family proteins can be differentially localized to microtubule plus ends during interphase and mitosis and that TACC is mainly required for recruitment of an XMAP215-family protein to interphase microtubule plus ends but not for recruitment to centrosomes and kinetochores. Moreover, we have now a marker to study dynamics and behavior of microtubule plus ends in living Dictyostelium cells. In a combination of live cell imaging of microtubule plus ends and fluorescence recovery after photobleaching (FRAP) experiments of GFP-α-tubulin cells we show that Dictyostelium microtubules are dynamic only in the cell periphery, while they remain stable at the centrosome, which also appears to harbor a dynamic pool of tubulin dimers.