Antigen uptake and immunoglobulin production in Atlantic cod (Gadus morhua L.) after intraperitoneal injection of Vibrio anguillarum.

Atlantic cod (Gadus morhua L.) were injected intraperitoneally with formalin-killed Vibrio anguillarum bacteria. Immunostaining revealed uptake of V. anguillarum antigens especially in the spleen after intraperitoneal (i.p.) administration. The uptake was time dependent in the interval 1-24 h. Most of the antigen uptake in the spleen was concentrated in areas around small blood vessels, while immunoglobulin producing cells were localised to some thick walled arteries. There was apparently little or no co-localisation of antigens and antibody producing cells. In the heart, some of the high endocardial endothelial cells of the atrium contained bacterial antigens and in head kidney some macrophage-like cells were stained. Very little antigen was found in the pigmented loose connective tissues of the peritoneum. In contrast, endothelial cells of the underlying blood vessels contained substantial amounts. In the heart, peritoneum and anterior kidney the number of antigen positive cells did not seem to change in the time interval 1-24 h. After i.p. immunisation with a mixture of V. anguillarum and Freunds complete adjuvant, the humoral immune response in Atlantic cod was low when tested 21, 42 and 105 days later. There was apparently no enhanced number of immunoglobulin synthesising cells caused by the antigen stimulation.

Expression of immunoglobulin heavy chain transcripts (VH-families, IgM, and IgD) in head kidney and spleen of the Atlantic cod (Gadus morhua L.).

Expression of the immunoglobulin (Ig) heavy chain transcripts in spleen and head kidney of the Atlantic cod (Gadus morhua L.) was investigated using in situ hybridization (ISH) and northern blotting. Specific detection of plasma cells was done with a probe for secretory IgM transcripts (mu 4). The plasma cells were often clustered close to blood vessels. Cells expressing surface IgM and IgD transcripts were detected using ISH with tyramide signal amplification (TSA). The positive cells were more abundant than plasma cells, had a lymphocyte-like morphology, and were evenly distributed throughout the tissues. This suggests that cod IgD mainly is expressed as a B-cell receptor akin to IgD in mammals. The VH-III family dominated the repertoire within the plasma cells, in agreement with data from cDNA cloning. Immunization with hapten-carrier antigen did not induce a systemic antibody response, and neither was any change in the clustering or distribution pattern of plasma cells within the tissues seen. A few clusters of plasma cells expressed only the rare VH-I and VH-II families, suggesting an ongoing clonal expansion and differentiation in these regions independently of immunization.

Ontogeny of lymphoid organs and immunoglobulin producing cells in Atlantic cod (Gadus morhua L.).

The ontogeny of lymphoid organs and the development of cells expressing immunoglobulin heavy chain (IgH) mRNA as well as cells containing immunoglobulin (IgM) were studied in Atlantic cod (Gadus morhua L.), a marine teleost. Head kidney and spleen appeared as the first lymphoid organs, present at the time of hatching, whereas thymus was observed in 9 mm larvae. Fully developed lymphoid organs were not achieved until after metamorphosis. Cells expressing IgH mRNA were detected in paraffin sections of larvae and juveniles by in situ hybridization. Positive cells were not detected in fish smaller than 33 mm (58 days after hatching). IgH mRNA expression coincided with the first appearance of immunoglobulin-positive cells as revealed by immunohistochemistry in the same animals.

Separation and determination of glycosaminoglycan disaccharides by micellar electrokinetic capillary chromatography for studies of pelt glycosaminoglycans.

Capillary electrophoresis based on cetyltrimethylammonium bromide micellar electrokinetic capillary chromatography (MECC) was developed for the separation and determination of glycosaminoglycan (GAG) disaccharide units without derivatization. The influence of changes in several separation conditions was studied, and the separation mechanisms are discussed. Tests of repeatability and linearity were performed for qualitative and quantitative evaluation of the method. The described procedure gives a rapid and efficient determination of GAG disaccharides. Samples of chondroitin sulphates and mink skin were treated with proteases, and the extent of protein cleavage was followed by free zone capillary electrophoresis. The result of the chondroitinase ABC treatment following the protease treatment was evaluated by the MECC method.

The effects of two mutations connected with chromatin functions on female germ-line cells of Drosophila.

We have studied the developmental effects of two dominant suppressor mutations of position-effect variegation mutations on female germ-line cells. Su-var(2)1(01), which has been shown to affect chromatin structure though altering histone deacetylation, and Su-var(3)3(03) are recessive female steriles and zygotic lethals in the presence of butyrate or an additional Y chromosome. We have analysed mosaic females with mutant germ-line and normal soma and concluded that intact functions of the Su-var(2)1 and the Su-var(3)3 genes are required for development of both the soma and the germ-line and that as indirect evidence suggest, their maternally provided products are needed for normal embryonic development. It is suggested that there is possibly a common control of chromatin structure and gene expression in the soma, female germ-line and embryonic cells of Drosophila.

Scanning electron microscopical observations on electrically fused human lymphocytes.

The paper describes scanning electron microscopic observations of electric field induced fusions of human lymphocytes. The cells were fused by application of short-time (20 microseconds) field pulses of 5 kV/cm after dielectrophoretic collection at about 1.2 MHz and 750 V/cm. Electron microscopy clearly demonstrates membrane fusion and the stability of initial stages of fusion.

Ultrastructural studies on plastids of generative and vegetative cells in Liliaceae : 5. The behaviour of plastids during pollen development in Chlorophytum comosum (Thunb.) Jacques.

The behaviour of plastids and mitochondria during the formation and development of the male gametophyte of Chlorophytum comosum has been investigated using electron microscopy. During first pollen mitosis an intracellular polarization of plastids occurs in that the plastids are clustered in the centre of the microspore. The originating generative cell normally lacks plastids. Only in a small number of microspores have plastids been observed near the dividing nucleus of the microspore and later on in the generative cell. These observations agree with the genetic investigations of Collins (1922) on the mode of plastid inheritance which demonstrated a small amount of biparental plastid inheritance in Chlorophytum. The cytological mechanisms underlying plastid polarization during the first pollen mitosis are discussed.