The switch from larval to adult globin gene expression in Xenopus laevis is mediated by erythroid cells from distinct compartments

The transition of hemoglobins during metamorphosis of Xenopus laevis involves replacement of the larval erythrocytes by adult ones, suggesting that the developmental control of this event depends upon the growth characteristics of the precursor cells. To identify the erythroid precursor cells and to investigate their developmental fate, we analyzed the distribution of stage-specific globin mRNAs by northern blotting in dorsal and ventral fragments of stage 32 embryos after in vitro culture as well as presumptive erythropoietic tissues of tadpoles during metamorphosis. The histological analysis shows that erythrocytes differentiate only in ventral fragments, suggesting that the ventral blood islands and most likely also the dorsolateral mesoderm are the primary sites of erythropoiesis. We also demonstrate that the first generations of erythrocytes, already express the predominating larval-specific alpha-globin mRNAs. The globin mRNA patterns obtained from presumptive erythropoietic tissues suggest an important role of circulating precursor cells in larval erythropoiesis, whereas the liver appears to be the main site of formation and maturation of the adult erythrocytes. Tentatively we propose that anuran erythropoiesis is dependent upon a self-perpetuating stem-cell line and that the larval and the adult erythrocytes are derived from successive generations of erythroid precursors, whose commitment may be imposed by the erythropoietic sites.     

Endoplasmic reticulum distribution during bovine oocyte activation is regulated by protein kinase C via actin filaments

Fertilization-induced [Ca²⁺]_i oscillations generally depend on the release of calcium ions from the endoplasmic reticulum (ER). Since ER is the main store of calcium ions, it plays an important role in oocyte fertilization. However, the mechanism of ER organization at oocyte activation is unknown. Here, we show that protein kinase C (PKC) is involved in ER distribution during bovine oocyte activation, but not involved in cell cycle resumption and spindle organization. Actin filaments were affected by PKC pharmacological inhibition. In addition, similar to PKC results, the actin-depolymerizing drug cytochalasin B affected the ER distribution during oocyte activation. Specifically, we have demonstrated that ER organization during bovine oocyte activation is regulated by PKC possibly through its action on actin filaments regulation. Taken together, the results presented here provide further information on the pathway involved in the regulation of ER organization during oocyte activation and new insight into the functional role of PKC and actin filaments during this process.     

Kinetics of the interaction of a 41-kilodalton macrophage capping protein with actin: promotion of nucleation during prolongation of the lag period

A 41-kilodalton macrophage capping protein (MCP) has been isolated which is capable of forming complexes with actin monomers in addition to capping the barbed ends of actin filaments (Southwick & DiNubile, 1986). The protein is calcium activated in a fully reversible manner. Using kinetic assays, we determined a capping constant, defined here as a modified Kd, of 1 nM and a Kd of 3-4 microM for MCP-actin monomer complex formation. MCP weakly nucleates actin polymerization: more than 0.5 microM MCP is necessary to shorten the lag period, and 1 microM MCP at an actin/MCP ratio of 10 reduces the average length of actin filaments to about 200 molecules per filament. We determined that the actin nucleus that survives MCP inactivation contains a minimum number of five actin molecules. These experiments also make a point with respect to the interpretation of the prolongation of the lag period. We directly demonstrate that in the presence of an actin binding protein a prolongation of the lag period can be associated with increased nucleation, contrary to the usual interpretation in the literature that it indicates no or decreased nucleation by the actin binding protein.     

Keratin-positive reticulum cells in fine needle aspirates and touch imprints of hyperplastic lymph nodes. A possible pitfall in the immunocytochemical diagnosis of metastatic carcinoma.

Fine needle aspirates and touch imprints of 36 hyperplastic (reactive) lymph nodes were tested for the presence of keratin and desmin. Keratin-positive cells with morphologic characteristics corresponding to extrafollicular (fibroblastic) reticulum cells were found in 18% of the fine needle aspirates and 42% of the touch imprints. The number of keratin-positive reticulum cells varied from 1 to greater than 30 per slide. Desmin-positive cells with similar morphology were found in 23% of fine needle aspirates and 37% of touch imprints, and the number of such cells per slide ranged from 2 to greater than 70. The relatively frequent occurrence of keratin-positive reticulum cells in these preparations from hyperplastic lymph nodes should be taken into account if keratin antibodies are used to search for carcinoma micrometastases.     

A recombination event excludes the ROM1 locus from the Best's vitelliform macular dystrophy region

Best's vitelliform macular degeneration has been genetically linked to chromosome 11. Subsequently, the disease locus has been refined to an interval between D11S903 and PYGM and, more recently, between D11S986 and D11S480. The gene encoding ROM1, a photoreceptor-specific membrane protein, has been independently mapped within the Best's disease region and has thus become a strong candidate for the Best's disease gene. In this study, we have mapped ROM1 relative to Best's disease and the loci D11S986, UGB (uteroglobin), and PYGM (human muscle glycogen phosphorylase) in recombinant Best's disease chromosomes. We demonstrate that UGB is localized proximal to ROM1 and that both UGB and ROM1 recombine with the disease phenotype. Thus, this analysis excluded ROM1 as the Best's disease gene.     

Advanced Liposomal Vectors as Cancer Vaccines in Melanoma Immunotherapy

Malignant tumors represent a major source of disability and account for more than one of five deaths in Western countries. Among the different cancers, melanoma harbors two distinctive features. First, its has long been recognized as an immunogenic tumor, and second, an unprecedented rise in incidence is currently observed, in face of few therapeutic options. Thus, melanoma represent an ideal target for a cancer immunotherapy program. To date, a number of immunodominant epitopes from tumor associated antigens (TAA) are used as cancer vaccines in clinical trials, in spite of an acknowledged rapid degradation in vivo and low immunogenicity. However, most of the immunotherapy trials reported so far do not achieve consistent clinical results. Hence, there is an urgent need for the development of a carrier system and strong adjuvants suitable for a TAA-based cancer immunotherapy. Liposomes and their further development as virosomes with added adjuvancy may address both these issues. We report here our experience in the tailoring of dedicated advanced liposomal vectors that were developed in the context of an upcoming immunotherapy clinical trial for melanoma.     

Amöboid bewegliche Pigmentzellen im Epithel des Seeigels Centrostephanus longispinus

Zusammenfassung Für den physiologischen Farbwechsel bei Vertebraten und Evertebraten gilt die Vorstellung, daß eine Pigmentbewegung innerhalb einer formkonstanten Zelle stattfindet. Am Seeigel Centrostephanus longispinus wird nun der Nachweis einer amoeboiden Bewegung von Pigmentzellen geführt: Die Epidermis von Centrostephanus enthält große braune Chromatophoren, die bei Belichtung eine Pigmentdispersion, bei Verdunkelung eine Konzentration des Pigments zeigen. Die Chromatophoren sind außerordentlich stark verzweigte Zellen, deren Arme dicht mit Pigmentgrana erfüllt sind. Im geballten Zustand ist die allgemeine Zellform mehr oder weniger ovoid, wobei die Zellarme eingezogen und dicht um die Zellmitte angeordnet sind. Dispersion des Pigments wird hervorgerufen durch Ausstrecken der pigmentierten Zellarme in den Interzellularraum des umgebenden Gewebes. Innerhalb der Zelle werden filamentöse Elemente nachgewiesen, die vermutlich für die Zellbeweglichkeit verantwortlich sind. — Ferner wird der zelluläre Aufbau des Integuments beschrieben.

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Amoeboid pigment cells in the epithelium of the sea urchin Centrostephanus longispinus A novel colour change mechanism

Summary Rapid colour changes in vertebrate and invertebrate species are considered to be due to movement of pigment granules within pigment cells of constant shape. Evidence is presented in this study to show that an amoeboid movement of chromatophores occurs in the epidermis of the Echinoderm Centrostephanus longispinus. The epidermis in this species contains large brown chromatophores, which display a dispersion of pigment on illumination and its concentration on darkening. The chromatophores are extensively branched cells, and their branches are densely packed with pigment granules. In the state of pigment concentration, the shape of the cell is more or less ovoid, and the cell branches are drawn in and closely arranged around the cell centre. Dispersion is attained by a stretching out of the pigmented cell branches into the intercellular spaces of the surrounding tissue. Within the cell, filamentous elements, which may be functional in the motility of the pigment cell, can be demonstrated.—Additionally the cellular composition of the integument is described.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft. Frl. A. Mikolaczick danken wir für sorgfältige technische Assistenz.     

„Elaioplasten“ fehlen den Schließzellen vonHosta plantaginea

Zusammenfassung Die Korollen-Epidermis vonHosta plantaginea besitzt in jungen Blüten in jeder Zelle je einen wohlentwickelten Elaioplasten, den Stomata-Schließ-zellen fehlen diese Inhaltskörper.