Ultrastructure of nervous tissue developing in the anterior chamber of the eye. 1. Perikaryons and dendrites

Embryonic tissues of septum and hippocampus were transplanted into the anterior eye chamber (AEC) of adult rats. The morphology of initial embryonic tissues and of transplants within 3 to 4 months of cultivation in AEC was studied. The transplanted tissue consists of neuroblasts and immature neurones: no synaptic contacts are observed. Within 3 to 4 months, highly differentiated neurones establishing synaptic contacts can be seen in the transplants. At the same time the fine structure of perikaryons and dendrites undergoes some changes: increased vacuolization, transformation of ergastoplasm into lamellar bodies. These can be due to an elevated functional activity of some neurones. Another group of morphological abnormalities (increased number of dendrite processes and microphyllopodia, somatic spines, dendrite cones of growth, tight junctions between perikaryons) suggests incomplete tissue maturation. These might be due to the absence of normal afferent and trophic influences in AEC.     

Ultrastructure of the synaptic endings in nerve tissue transplants developing in the anterior chamber

Embryonic septum of hippocampus was grafted into the anterior eye chamber (AEC) of adult recipient rats. The fine structure and distribution of synaptic endings were studied in the hippocampus (HC) and septum (ST) grafts developing in oculo for 3-4 months. On the basis of the structure of postsynaptic regions, asymmetrical and symmetrical synapses are distinguished, whose distribution on the body and dendrites of hippocampal and septal neurons is basically similar with that in situ. As in vivo, axo-somatic, axo-dendrite and axo-spine forms of synaptic endings have been observed. Neuropile has, basically, normal structure, judging by the ratio of nerve and glial elements, but sometimes dendro-dendrite contacts and glomerular-like synaptic structures are observed which are not characteristic of the studied brain regions. Besides, the grafts contain an increased number of serial and tangential synapses, as well as axonal terminals with the signs of growth cones. The observed structural deviations appear to be due to incomplete tissue maturation in the absence of normal afferentation.     

Mutagenic effect of mitomycin C on different strains of oleandomycin producer Streptomyces antibioticus

Mitomycin C, a DNA-tropic antibiotic, was shown to have a lethal effect on spore sprouts of two strains of Streptomyces antibioticus, an organism producing oleandomycin. When the time of exposure to the antibiotic increased there was an almost equal decrease in the survival rate. The mutagen action on the morphological variation and antibiotic production of the two closely related strains were diverse due to their genetic differences. The strain isolated after the culture treatment with a chemical mutagen and subjected to a more prolonged maintaining selection showed lower variation with respect to its colony morphology. The other strain isolated after treatment of the culture with high concentrations of its own antibiotic showed lower variation with respect to its antibiotic production property. The shift in the antibiotic production in the direction of the low active variants was characteristic of the both highly productive strains.     

Use of transposons for studying the genetic organization of Vibrio cholerae non 01]

The nonagglutinating vibrions having Tn-elements inserted into the chromosome were obtained as a result of conjugal transfer of vector plasmids carrying the different transposons (Tn9, Tn10, Tn601, Tn5-Mob) and of the consequent isolation of plasmid-free clones of Vibrio cholerae non OI. Identification of auxotrophic mutations induced by the transposons inserted into the bacterial genome made possible the construction of the primary chromosomal map of Vibrio cholerae non OI. The efficient donor strains of Vibrio cholerae non OI were constructed by introducing the transposon Tn5-Mob and the helper plasmid RP-4. The donors are capable of oriented conjugal transfer of chromosome.     

Transposon mutagenesis, elimination and mobilization of plasmids in nitrogen-fixating bacterium Azospirillum brasilense Sp245

The expressed difference in the plasmid profile of A. brasilense Sp245 is registered as a result of Tn5-Mob-mutability. Integration of the vector pSUP5011 into one of the A. brasilense Sp245 plasmid and using of the Tn5-Mob transposon to mobilize the 85Md cryptic plasmid are reported. The properties of A. brasilense Sp245 with the mutant plasmids composition (surface structure, acetylene and nitrate reduction, ability to a number of carbohydrates utilization, formation of melanin, antibiotics resistance specter) have been analyzed. The transposon Tn5-Mob insertion into the 85Md plasmid resulted in isolation of a mutant excreting a melanin-like pigment into the medium. The results suppose 85Md plasmid participation in melaninogenesis.     

Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections

A method of rapid freezing in supercooled Freon 22 (monochlorodifluoromethane) followed by cryoultramicrotomy is described and shown to yield ultrathin sections in which both the cellular ultrastructure and the distribution of diffusible ions across the cell membrane are preserved and intracellular compartmentalization of diffusabler ions can be quantitated. Quantitative electron probe analysis (Shuman, H., A.V. Somlyo, and A.P. Somlyo. 1976. Ultramicros. 1:317-339.) of freeze-dried ultrathin cryto sections was found to provide a valid measure of the composition of cells and cellular organelles and was used to determine the ionic composition of the in situ terminal cisternae of the sarcoplasmic reticulum (SR), the distribution of CI in skeletal muscle, and the effects of hypertonic solutions on the subcellular composition if striated muscle. There was no evidence of sequestered CI in the terminal cisternae of resting muscles, although calcium (66mmol/kg dry wt +/- 4.6 SE) was detected. The values of [C1](i) determined with small (50-100 nm) diameter probes over cytoplasm excluding organelles over nuclei or terminal cisternae were not significantly different. Mitochondria partially excluded C1, with a cytoplasmic/ mitochondrial Ci ratio of 2.4 +/- 0.88 SD. The elemental concentrations (mmol/kg dry wt +/- SD) of muscle fibers measured with 0.5-9-μm diameter electron probes in normal frog striated muscle were: P, 302 +/- 4.3; S, 189 +/- 2.9;C1, 24 +/- 1.1;K, 404 +/- 4.3, and Mg, 39 +/- 2.1. It is concluded that: (a) in normal muscle the "excess CI" measured with previous bulk chemical analyses and flux studies is not compartmentalized in the SR or in other cellular organelles, and (b) the cytoplasmic C1 in low [K](0) solutions exceeds that predicted by a passive electrochemical distribution. Hypertonic 2.2 X NaCl, 2.5 X sucrose, or 2.2 X Na isethionate produced: (a) swollen vacuoles, frequently paired, adjacent to the Z lines and containing significantly higher than cytoplasmic concentrations of Na and Cl or S (isethionate), but no detectable Ca, and (b) granules of Ca, Mg, and P = approximately (6 Ca + 1 Mg)/6P in the longitudinal SR. It is concluded that hypertonicity produces compartmentalized domains of extracellular solutes within the muscle fibers and translocates Ca into the longitudinal tubules.     

Argasid tick lysozyme action on HEp-2 cells

The cytotoxic effect on Argasidae lysozyme was shown on a model of cell line HEp-2 in comparison to egg lysozyme. The lysozyme was obtained from homogenates of Ornithodoros papillipes of subfamily Ikodoidea. The lysozyme in concentrations of 300 and 500 gamma/ml had a cytotoxic effect, while in doses of 50 and 100 gamma/ml it has no such activity. The cells of line FL were not sensitive to the above concentrations of the lysozyme. In concentrations of 500 gamma/ml and higher the egg lysozyme had an analogous cytotoxic effect on the cells of line Hep-2. The comparative study of the cariogrammes of the cell monolayer treated with the Argasidae and egg lysozymes, as well as the study of the level of 3H-thimidine incorporation into the cell DNA showed the absence of the preparations effect of the mitotic activity of the cells and DNA synthesis by them.     

Production in an Actinomyces levoris culture under the action of an actinophage specific to it of variants with stable preservation of its resistance to the phage]

Act. levoris 28, an organism producing levorin was treated with an actinophage virulent to it. Variants of the organism were isolated from the secondary growth of the culture. As a result of lysogenization with the above phage the variants acquired stability to it which was preserved during the further generations. In the previous experiments carried out by the authors the variants isolated from the secondary growth of the culture after its exposure to the same phage lost their stability to the phage as a result of loosing the prophage by it during the subsequent passages. The phage stable variants did not differ from the initial culture either in the activity of levorin or the levorin composition. The phages found in the initial culture 28, and the virulent mutant were identical with respect to the particles morphology and antigenic properties which confirmed their relation.