Accumulation of a peptide toxin from the cyanobacterium Oscillatoria agardhii in the freshwater mussel Anadonta cygnea

Swan mussels (Anodonta cygnea) were exposed to a toxic strain of the cyanobacterium Oscillatoria agardhii. Mussels accumulated large amounts of the peptide Oscillatoria toxin which was present in low concentrations within the cyanobacterial cells in the test aquaria (40–60 µg Oscillatoria toxin/1). The toxin concentration in the mussels increased during the experiment and after 15 days of exposure the concentration was 70 ± 2 µg/g freeze dried tissue (mean ± range of values). The highest concentration of the toxin (130 µg/g of freeze dried tissue) was found in the hepatopancreatic tissue. The toxin did not seem to be metabolized in the mussels and they were not killed by the high toxin concentrations within them. After two months in clean water still detectable amounts of toxin were present in the mussels.     

Assignment of the gene encoding the catalytic subunit Cβ of CAMP-dependent protein kinase to the p36 band on chromosome

Summary A cDNA for the human catalytic subunit (C) of cAMP-dependent protein kinase (PKA) has been cloned from a testis cDNA library. In the present study, we have determined the chromosomal localization of this gene using a cDNA for C as a probe. Southern blot analysis of genomic DNA from human/mouse cell hybrids revealed that the presence or absence of a 20-kbXbaI fragment, which hybridized with the C probe, was concordant with the presence of human chromosome 1.In situ hybridization to metaphase chromosome confirmed the somatic cell hybrid data and regionally mapped the C gene of PKA to the p36 band on chromosome 1.     

Variation in plumage colour of the Great tit Parus major in relation to habitat, season and food

The plumage colour of nestling and adult Great tits Parus major major L. was studied at Trondheim (63 °N, 10 °E), Norway. Nestlings reared in deciduous woodland were yellower than those reared in coniferous woodland, those reared in 1983 were yellower than those in 1982, and the later in the breeding season they were reared the more yellow they were. Furthermore, adults were yellower than nestlings, and for both groups the males were yellower than the females. Experimental interchange of eggs between nests showed that the variation in plumage colour observed between broods was related primarily to the food and not to genetic differences. Sampling, by the neck-collar method, of the food brought by the parent birds showed that the yellower nestlings received a greater proportion of lepidopteran larvae, in particular greenish-coloured ones, than did the nestlings with a paler plumage. A speculation is whether or not the variation in plumage colour has some adaptive value (viz. background matching).
In avian systematics, plumage coloration has been used to distinguish and describe subspecies. The present study shows that such a classification should only be made with certain reservations, until possible variation in the food quality of the subspecies is known to have no influence on plumage coloration.     

The localization of a vitamin K-induced modification in an N-terminal fragment of human prothrombin

1. The N-terminal fragment (PF-I) split off from prothrombin during coagulation was purified to homogeneity from human serum. 2. The apparent molecular weight is 27000+/-2000 in sodium dodecyl sulphate-polyacrylamide-gel electrophoresis, whereas a value of about 19600 is obtained by calculation based on amino acid and carbohydrate analyses. The N-terminal sequence is an Ala-Asx bond. The fragment contains about 16% carbohydrate, binds phospholipids in the presence of Ca(2+) and is adsorbed to BaSO(4). The pK(a) of its BaSO(4)-binding group(s) is 3.1-3.5. 3. By CNBr cleavage of fragment PF-I two peptides (C-1 and C-2) were obtained with molecular weights of about 5900 (C-2) and 12400 (C-1) on the basis of amino acid and carbohydrate analyses. Only the smaller (N-terminal) peptide is adsorbed to BaSO(4) and, since the ability of the whole protein to bind to BaSO(4) is known to be absent in samples obtained from patients treated with vitamin K antagonists, this peptide probably contains the site of a modification to the structure of the protein which occurs during biosynthesis and depends on vitamin K. This peptide does not contain hexosamine or sialic acid.     

Disintegration of H3-labelled spermatocytes in Melanoplus differentialis

Summary Whole cysts in the testicular tubes of Melanoplus differentialis disintegrate giving rise to large Feulgen positive bodies. The disintegration affects all the nuclei of a cyst.The Feulgen positive bodies are strongly labelled with tritiated thymidine, a confirmation of their DNA content.Spectrophotometric measurements reveal that the bodies contain 3.5 times more DNA per unit area than the autosomes of normal spermatocyte nuclei and an equal amount of DNA per unit area as the sex chromosome of the same nuclei.This regular disintegration of spermatocytes is not considered as a pathological condition but as an adaptation by which large amounts of DNA are easily released at a convenient time of development.Supported by a research grant from the Swedish Natural Science Research Council.     

The Gene Encoding the Catalytic Subunit Cα of cAMP-Dependent Protein Kinase (Locus PRKACA) Localizes to Human Chromosome Region 19p13.1

We have determined the chromosomal localization of the gene for the catalytic subunit Cα of cAMP-dependent protein kinase (locus PRKACA) to human chromosome 19 using polymerase chain reaction (PCR) and Southern blot analysis of two different somatic cell hybrid mapping panels. In addition, PCR analysis of a chromosome 19 mapping panel revealed the presence of a human Cα-specific amplification product only in cell lines containing the region 19p13.1 to 19q12. Finally, two-color fluorescencein situhybridization to metaphase chromosomes using the human Cα cDNA and human chromosome 19 inter-Alu-PCR product as probes localized the human Cα gene to chromosome region 19p13.1.     

The Use of Hydrophobic Adhesive Tape to Produce Miniature Wells on Microscope Slides

Hydrophobic adhesive tape was used to produce miniature wells on microscope slides for staining several sections of tissue with minimal amounts of cytochemical reagents. The wells could be tailored to individual specifications and the method allowed coverslips to be mounted close to the sections using either aqueous or xylene based mounting media. This method was especially useful for multiple immunolabelling of serial semithin cryosections.     

Electron microscopy of the tracheal ciliated mucosa in rat

Summary The structure of the tracheal epithelial cells from rat has been studied by electron microscopy on approximately 200 Å thick sections with a resolution of better than 30 Å.The epithelium is found to be of a simple columnar type composed of ciliated cells, mucus producing (goblet) cells, basal cells and a fourth kind of cell, here called brush cell. A great number of non-ciliated cells has also been encountered. It has been proved that these represent goblet cells in different stages of intracellular synthesis of mucous granules. The ciliated cells have approximately 8–9 cilia per square micron and there are about 270 cilia on each cell, the calculated surface area being 33 square microns. They are covered by a 70 Å thick membrane. The ciliary filaments are arranged in a pattern of 2 separate ones in the center and a ring of 9 peripheral ones, each divided into 2 subfilaments by a wall with same thickness as the filamentous wall itself, this being 60 Å. The peripheral filaments are continuous with the basal corpuscles. The structure of the corpuscles as compared with earlier findings is discussed. A number of 0.05 micron thick and 1 micron long filiform projections emerge from the cell surface. No cuticle is present.The cell membrane facing adjacent cells is 90 Å and separated from their cell membrane by a 105 Å wide space, this space, being expanded towards a level corresponding to the proximal parts of the cell. A structure that represents terminal bar has been encountered. The cytoplasm is loose and composed of 160 Å thick granules. Spaces enclosed by 50 Å thick membranes with attached 160 Å thick granules (-cytomembranes) are rare. The Golgi zone is analyzed and its regular composition of -cytomembranes, granules and vacuoles is confirmed. The mitochondria with a mean width of 0.23 micron differ to their inner structure from the common type in that the triple layered membranes are highly interconnected. Large opaque granules are encountered in the cytoplasm. Ring-shaped, 850 Å wide, structures are present in the nuclear membrane. The goblet cells are not as abundant as the ciliated cells, the ratio being 14. Small filiform projections covered by a 95 Å thick membrane protrude from the cell surface. This membrane is continuous with the cell membrane, the latter with the same dimensions as in the ciliated cells. Terminal bars are present. The cytoplasm is very opaque due to a dense packing of the 165 Å opaque granules, many in clusters of 4–6. The -cytomembranes have the same dimensions as mentioned above for those present in the ciliated cells. The Golgi zone is of regular composition. There is a suggestion that the Golgi vacuoles and the -cytomembranes are involved in the formation of mucus. In the stage of cellular activity with but few mucous granules, there is a great number of large opaque granules, the size varying from 0.4–1 micron. The mitochondria with a mean width of 0.23 micron have an outer triple layered membrane with a total thickness of 180 Å. The central less opaque layer is 70 Å and the opaque layer on either side is 55 Å. The inner membranes are arranged parallel to each other and have a triple layered composition where the central less opaque layer is 65 Å and the opaque layers each 60 Å. The brush cells belong to the non-ciliated cells. They are encountered singly, surrounded by goblet cells. The surface structures are shaped like brushes or clumsy protrusions which emerge from the distal end of the cell, and are covered by a 95 Å thick membrane. There have been no suggestions of the brushes being cilia in a stage of growth, nor is it probable that they represent stereocilia. They most nearly resemble the intestinal brush border extensions and thus might serve as a resorbing structure.The cytoplasm of the brush cells appears of medium opacity between the ciliated cells and goblet cells and is composed of 155 Å opaque granules. The -cytomembranes are very rare. The Golgi zone is diminutive though of regular composition. The mitochondria are abundant and small with a mean width of 0.14 micron. The outer and inner membranes are triple layered with approximately the same dimensions as reported for the mitochondria of the ciliated and goblet cells. The inner membranes are very few, often only one or two are present. Some of the large opaque granules have inside a very regular arrangement of small 60 Å thick opaque granules arranged in a crystallinic pattern. In the cytoplasm 0.5–1 micron long bundles of 30–40 Å wide fibrils are encountered. The nucleolus shows a characteristic structure of concentrically arranged thin membranes. The basal cells are believed to represent lymphocytes or white blood cells. They sometimes rest on the basement membrane, sometimes are encountered in the distal part of the intercellular spaces. They are bordered by a 110 Å thick cell membrane and have a rather opaque cytoplasm characterized by 160 Å thick opaque granules. A very small Golgi zone is present. The mitochondria, the mean width being 0.14 micron, have triple layered outer and inner membranes, where the less opaque central layer is 65–70 Å and the opaque layers 45–50 Å each. The basement membrane has a thickness of 600 Å. No inner structure has been resolved. The basement membrane is separated from adjacent parts of the ciliated, goblet, brush, and basal cells by a 250 Å wide less opaque space. Below the basement membrane is the lamina propria of the trachea, which is composed of collagen and elastin fibers together with fibroblasts, white blood cells and lymphocytes. The relationship between different types of tracheal epithelial cells in rat has been analyzed. There has been found no indication of a transformation of any type of cells observed into a different type of cell. The development of basal cells via supporting cells or intermediate cells to goblet cells or ciliated cells has not been noticed. On the contrary, all cells that in light microscopy could have been considered to be supporting or intermediate cells, we have been able to recognize as brush cells or as goblet cells to a varying degree filled with mucous granules. If the cells did not seem to reach the cell surface it has been found to be due to a diagonal direction of the sectioning. In this connection it should be emphasized that this relationship is valid only in rat where it is known that the epithelium is of a simple columnar type as distinct from the conditions in man, that epithelium being of a pseudostratified columnar type.This paper is based on a report given at the meeting of Deutsche Gesellschaft für Elektronenmikroskopie in Münster, March 28–31, 1955 and at the Scandinavian Electron Microscope Society Meeting in Stockholm, May 13, 1955.