Shortening velocity of single muscle cells isolated from a molluscan smooth muscle

Summary The maximal unloaded shortening velocity (V_max) of smooth muscle cells isolated from the pedal retractor muscle ofMytilus was more than twice as large as that of the whole muscle, suggesting the presence of extracellular components which resist the contraction of the whole muscle. The V_max of the isolated cells was almost constant at cell lengths ranging between 0.5 and 0.831₀ (1₀, optimal length for tension generation) indicating that the intracellular resistance to contraction is negligible within this range of lengths.     

Structural characteristics and distribution of satellite cells along crayfish muscle fibers

The distribution of satellite cells (sc) in long-sarcomere muscle fibers from the carpopod extensor muscle of the crayfish (Astacus fluviatilis) has been studied electron-microscopically. The sc are spindle-shaped and are oriented parallel to the long axis of a fiber. The mean lengths of sc nuclei (17.00 m) and that of myonuclei (18.35 m) differ non-significantly. In older animals, the mean ratio of the number of sc nuclei to the total number of nuclei (sc nuclei + myonrclei) is 0.0716, 0.0848, and 0.034 for the tendon, central and shell segments, respectively. The corresponding values for younger animals are 0.158, 0.166, and 0.081. The mean numbers of sc nuclei per mm of a fiber are 94, 117, and 47 (older animals), and 164, 117, and 94 (younger animals) for the tendon, central and shell segments, respectively. The high incidence of sc per unit fiber length in crayfish may be related to the fact that crayfish muscle fibers have a much larger diameter than vertebrate muscle cells.     

Serotonin-stimulated protein phosphorylation in aortic smooth muscle cells

Summary The effects of serotonin on the formation of inositol phosphates and protein phosphorylation were examined in cultured smooth muscle cells. Serotonin stimulated the formation of [³H]inositol monophosphate, [³H]inositol bisphosphate and [³H]inositol trisphosphate. This effect was prevented by 5-HT₂ specific antagonist, 6-methyl-1-(1-methylethyl)ergoline-8-carboxylic acid, 2-hydroxy-1-methylpropyl ester [Z]-2-butenedioate (LY53857). Serotonin stimulated the phosphorylation of many polypeptides, among which a 20 kDa polypeptide was the most prominent. The phosphorylation was also inhibited by LY53857. LY53857 alone produced no effects on protein phosphorylation. The 20 kDa polypeptides were also phosphorylated by the addition of 12-O-tetradecanoylphorbol-13-acetate. These results suggest that serotonin stimulates protein phosphorylation through 5-HT₂ receptors and possibly activates protein kinase C in intact vascular smooth muscle cells.Part of the data contained in this paper was presented at the 74th local meeting of the Japanese Society of Pharmacology at Kanagawa.     

Retinoic acid enhances the proliferation of smooth muscle cells

Summary Retinoic acid (RA, 10^–5–10^–7 M) is shown to enhance the proliferation of cultured rat aortic smooth muscle cells (SMC). This effect is not connected with a synergistic action of RA together with serum mitogens. Moreover, the expression of L1, a surface antigen specific for modulated SMC entering the cell cycle, is amplified by RA treatment.     

Communicating junctions between pillar cells in the gills of the atlantic hagfish,Myxine glutinosa

Summary In the respiratory lamellae of the gills of the atlantic hagfish,Myxine glutinosa, pillar cells are coupled by communicating junctions. It is suggested that these cells behave like a functional syncytium.Supported by the DFG, SFB 146. The authors are indebted to Drs F. Walvig and H. Stolte for kindly providing the hagfish. The expert technical assistance of G. Voss-Wermbter, Dipl.-Ing., and H. Heidrich is gratefully acknowledged.     

Ciliated fibroblasts and smooth muscle cells in the rat uterus

Summary Ciliation in endometrial fibroblasts and myometrial muscle cells of the rat was examined by transmission electron microscopy. Quantification of the number of ciliated cells during the estrus cycle did not show any firm relationship between cilation and ovarian hormonal activity. In the case of most cilia, there is a spatial relationship between their basal centrioles and the Golgi complex, so that a Golgi-cilium complex is created. A possible role of ciliation in uterine fibroblasts and smooth muscle cells is discussed.     

Colchicine-induced appearance (proliferation) of smooth sarcoplasmic reticulum in arterial smooth muscle cells

Summary Colchicine treatment resulted in the appearance and proliferation of smooth sarcoplasmic reticulum in some smooth muscle cells of the aortic and pulmonary trunk walls in the rabbit. The significance of cytoplasmic microtubules and/or membrane-bound tubulin for the morphogenesis, functioning and control of smooth endoplasmic reticulum in different kinds of cells is discussed.     

Partial purification of an endogenous elicitor from suspension-cultured cells of red bean,Vigna angularis

Brief irradiation with a germicidal UV lamp of cells of red bean,Vigna angularis, cultured in suspension in a quartz flask caused the release into the culture medium of an endogenous substance with elicitor activity, as well as the accumulation of isoflavone glucoside stress metabolites in the cells. The active compound was fractionated using phenylalanine ammonia lyase (PAL)-inducing activity in fresh cells as a marker. The elicitor active principle appears to be a low molecular weight (<2000 MW) water-soluble acidic oligosaccharide.     

Effect of aloe lectin on deoxyribonucleic acid synthesis in baby hamster kidney cells

Summary A homogeneous glycoprotein (mol.wt 40,000) containing 34% carbohydrate was isolated fromAloe arborescens var.natalensis. At a concentration of 5 g/ml, this glycoprotein was shown to stimulate deoxyribonucleic acid (DNA) synthesis in baby hamster kidney (BHK) cells and to have the properties of a lectin which reacts with sheep blood cells. The chemical and physical properties of the glycoprotein (aloe lectin) are also discussed.     

Dynamics of increase in muscle fibers in fishes in relation to size and growth

Summary Dynamics of increase of white myotomal muscle fibers of four species of freshwater teleosts (Salmo gairdneri, Pimephales notatus, Esox masquinongy andE. americanus vermiculatus) from three families (Salmonidae, Cyprinidae, Esocidae) representing a variety of maximum attainable sizes and growth rates, have been investigated. There are at least three major differences in these dynamics, and there appears to be an association between the ability of a fish species to attain large size (and grow fast) and its ability to recruit new fibers into this predominant tissue of the myotomal mass.     

The potency of heparin-like activity of glycosaminoglycans released by human endothelial cells in culture

Summary Glycosaminoglycans isolated from culture medium conditioned by human endothelial cells showed heparin-like antithrombin III cofactor activity measured by Xa inhibition. Their activity was relatively weak, 0.1% of the potency of heparin, but was approximately 5-fold more potent than that of glycosaminoglycans derived from vascular smooth muscle cells.     

Sphingosine 1-phosphate induces differentiation of adipose tissue-derived mesenchymal stem cells towards smooth muscle cells

Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid which regulates multiple biological parameters in a number of cell types, including stem cells. Here we report, for the first time, that S1P dose-dependently stimulates differentiation of adipose tissue-derived mesenchymal stem cells (ASMC) towards smooth muscle cells. Indeed, S1P not only induced the expression of smooth muscle cell-specific proteins such as α-smooth muscle actin (αSMA) and transgelin, but also profoundly affected ASMC morphology by enhancing cytoskeletal F-actin assembly, which incorporated αSMA. More importantly, S1P challenge was responsible for the functional appearance of Ca²⁺ currents, characteristic of differentiated excitable cells such as smooth muscle cells. By employing various agonists and antagonists to inhibit S1P receptor subtypes, S1P₂ turned out to be critical for the pro-differentiating effect of S1P, while S1P₃ appeared to play a secondary role. This study individuates an important role of S1P in AMSC which can be exploited to favour vascular regeneration. Received 06 March 2009; accepted 17 March 2009     

Satellite cells in the regenerated and regrafted skeletal muscles of rats

Summary Soleus (SOL) muscles were grafted into extensor digitorum longus (EDL) muscle beds (EDL-first-graft). Sixty days later, some mature EDL-first-grafts were regrafted into their own beds (EDL-second-grafts). Fully regenerated muscle fibers and satellite cells were observed in both types of mature grafts. The ratios of satellite cell nuclei per total nuclei (myonuclei and satellite cell nuclei) were 4.81±0.47% for EDL-2nd graft, 4.26±0.51% for EDL-1st-graft, 4.30±0.33% for control SOL, and 3.30±0.18% for control EDL. It is thought that satellite cells are required for the repeated activity of muscle fiber regeneration. The persistance of satellite cells in EDL-second-grafts suggests that satellite cells are not depleted during the first grafting, making second-grafts possible.     

Allatotropic activity of the median neurosecretory cells ofGalleria mellonella (lepidoptera) larval brain

Summary The brain allatotropic hormone (ATTH) is released inGalleria mellonella from the Median neurosecretory cells located in the pars intercerebralis. These cells show the ability to elicit supernumerary larval molts upon implantation into sensitive host larvae, and the ability to in vitro stimulate the juvenile hormone synthesis in corpora cardiaca-corpora allata glands ofG. mellonella.11 November 1986Acknowledgments. I am greatly indebted to Wesleyan University (Connecticut, USA) for supporting this study.     

Legionella pneumophila down-regulates MHC class I expression of human monocytic host cells and thereby inhibits T cell activation

Legionella (L.) pneumophila, the causative agent of Legionnaires disease, is an intracellular pathogen of alveolar macrophages that resides in a compartment displaying features of endoplasmatic reticulum (ER). In this study, we show that intracellular multiplication of L. pneumophila results in a remarkable decrease in MHC class I expression by the infected monocytes. During intracellular multiplication, L. pneumophila absorbs ER-resident chaperons such as calnexin and BiP, molecules that are required for the correct formation of the MHC class I complex. Due to reduced MHC class I expression, stimulation of allogeneic blood mononuclear cells was severely inhibited by infected host cells but cytotoxicity of autologous natural killer cells against Legionella-infected monocytes was not enhanced. Thus, reduced expression of MHC class I in infected monocytes may resemble a new immune escape mechanism induced by L. pneumophila.Received 22 November 2004; received after revision 27 December 2004; accepted 5 January 2005     

2-Carboxyethylgermanium sesquioxide,a synthetic organogermanium compound,as an inducer of contrasuppressor T cells

2-Carboxyethylgermanium sesquioxide (Ge-132), a synthesized organogermanium compound with immunomodulaing activities, was shown to be an inducer of anti-suppressor T cells in normal mice. The suppressor cell activity of T6S cells, a clone of burn-induced CD8⁺ IL-4-producing suppressor T cells, was clearly inhibited when a mixed lymphocyte-tumor cell reaction of the clone was conducted with splenic mononuclear cells from mice treated orally with a 100 mg/kg dose of Ge-132. The activity of anti-suppressor cells was demonstrated in spleens of mice 2 days after treatment with Ge-132 and reached its peak on day 3. The anti-suppressor cells induced by the compound were of a contrasuppressor T cell-linage, because they were characterized as CD4⁺ CD28⁺ TCR/⁺ Vicia villosa lectin-adherent T cells. These cells produced IFN- but did not produce IL-2, IL-4, IL-6 or IL-10 in their culture fluids. CD4⁺ anti-suppressor T cells induced by Ge-132 may be different from other subsets of CD4⁺ T cells because Th1 and Th2 cells generated in our laboratory did not adhere toVicia villosa lectin-coated petri dishes, and each produced specific cytokines. Th1 cells produced IFN- and IL-2 while Th2 cells produce IL-4 and IL-10 in vitro. These results suggest that Ge-132 may be useful as an inducer of contrasuppressor T cells in immunocompromised individuals bearing suppressor T cells. To eliminate suppressor T cells from immunocompromised hosts may result in improved resistance from various opportunistic infections.     

Selective killing of smooth muscle cells in culture by the ricin A-chain conjugated with monoclonal antibodies to a cell surface antigen via a dextran bridge

Summary Monoclonal antibodies to a surface antigen of the modulated smooth muscle cells originally isolated from the rat aorta media were conjugated with ricin A-chain via an oxidized dextran bridge. The interaction of cultured cells with the conjugates obtained and with control substances was monitored following incorporation of¹⁴C-leucine radioactivity. It was found that¹⁴C-leucine incorporation was suppressed by 80–90% at a conjugate concentration of 10^–6–10^–7 M. Antigen-negative cells (line IAR; rat hepatocytes) were insensitive to the conjugate at any concentration used. Control use of purified ricin A-chain, native or oxidized dextran, specific and nonspecific IgG did not affect normal¹⁴C-leucine incorporation. The data obtained may be useful for designing targeted drug transport systems and for selective screening of modulated smooth cells in vascular pathology models in vivo.     

Microtubule inhibitors block the morphological changes induced inDrosophila blood cells by a parasitoid wasp factor

Summary The shape change ofDrosophila melanogaster blood cells (lamellocytes) from discoidal to bipolar that is caused by a factor from the female parasitoidLeptopilina heterotoma is blocked by the tubulin inhibitors vinblastine and vincristine in vitro. The actin inhibitor, cytochalasin B, causes arborization ofDrosophila lamellocytes and acts synergistically with the wasp factor to alter lamellocyte morphology. Lamellocyte arborization induced by cytochalasin B is blocked by simultaneous treatment with vinblastine. These observations indicate that the changes in lamellocyte shape induced by both the wasp factor and cytochalasin B require microtubule assembly.     

DNG1, a Dictyostelium homologue of tumor suppressor ING1 regulates differentiation of Dictyostelium cells

dng1 is a Dictyostelium homologue of the mammalian tumor suppressor ING gene. DNG1 protein localizes in the nucleus, and has a highly conserved PHD finger domain found in chromatin-remodeling proteins. Both dng1 disruption and overexpression impaired cell proliferation. In dng1-null cells, the progression of differentiation was delayed in a cell-density-dependent manner, and many tiny aggregates were formed. Exogenously applied cAMP pulses reversed the inhibitory effect caused by dng1 disruption on the aggregation during early development, but formation of tiny aggregates was not restored. dng1-overexpressing cells acquired the ability to undergo chemotaxis to cAMP earlier and exhibited enhanced differentiation. These phenotypes were found to be coupled with altered expressions of early genes such as cAMP receptor 1 (car1) and contact site A (csA). Furthermore, disordered histone modifications were demonstrated in dng1-null cells. These results suggest a regulatory role of dng1 in the transition of cells from growth to differentiation.Received 29 December 2004; received after revision 24 May 2005; accepted 26 May 2005