Preparative thin-layer and column chromatography of prostaglandins

Chromatographic methods are used to identify various monounsaturated PGs (prostaglandins). TLC (thin-layer chromatography) and column chromatography are described in detail. These systems were developed specifically for separating epimers of PGE1 and PGF1, but they are useful in separating some of the known natural PGs as well. A table presents information on identification of the various PGs with neutral silica and acidic silica. The results of these laboratory experiments indicate that 1 ug-1g of PG can be purified by chromatographic methods with little or no loss due to irreversible adsorption or rearrangement if proper precautions and solvent systems are used. TLC seems to be useful in distinguishing diastereomers of PGs. Relative mobilities of the various hydroxy diastereomers are not changed by the solvent systems or by esterification.     

HETEROGENEOUS DISTRIBUTION OF ENZYMES IN SUBMICROSOMAL MEMBRANE FRAGMENTS

Microsomal membranes are postulated to contain either a homogeneous arrangement of individual enzymes or groupings of functionally related enzymes. In the present study we attempt to distinguish between these hypotheses in subfractions of rough microsomes from rat liver. After sonication, the individual vesicles that make up the rough-membrane fraction average less than 1/100 of their previous mass. The vesicles in the sonicated suspension are fractionated roughly according to size on a continuous sucrose gradient. Enzyme activity or concentration in fractions of the gradient is expressed on a phospholipid basis. Fractions containing primarily small vesicles differ from those containing larger vesicles in a manner suggesting a certain degree of separation of NADH-linked from NADPH-linked enzymes. NADH-ferricyanide reductase, NADH-cytochrome c reductase and cytochrome b₅ are most concentrated within the large vesicles in the lowest third of the gradient. In contrast, NADPH-cytochrome c reductase and cytochrome P-450 are found in highest concentration in the small vesicles that make up the upper third of the gradient. The results suggest a nonrandom distribution of these two enzyme groups in the membranes of the endoplasmic reticulum.     

Electron microscopic observations on iron-labelled secondary lysosomes in renal tubules in mice

Summary Labelling of renal tubule cytosomes with electron dense iron granules can be attained by daily intramuscular injections to mice of an iron sorbitol citric-acid compound in a total of approximately 50 mg Fe⁺⁺⁺/100 g of body weight. The labelled cytosomes correspond to secondary lysosomes and represent heterolysosomes or ambilysosomes. The evidence suggests that tubule and lysosome function are undisturbed by the labelling procedure. The use of this method for fine structural studies of the interaction between secondary lysosomes and other cytoplasmic organelles and elements is indicated.Microbodies do not incorporate administered Fe⁺⁺⁺. The morphological observations support the opinion that these bodies are formed in specialized portions of the smooth surfaced endoplasmic reticulum of proximal tubule cells.Supported in part by grants from the Swedish Medical Research Council (Projects No. K 67-12x-1006-2, B 67-12x-1006-02K, K 68-12x-1006-03, and B 69-12x-1006-04A). The assistance of Miss Silwa Mengarelli and Miss Britt-Marie Pettersson is gratefully acknowledged.     

The appearance of monoamines in the adult and developing neurohypophysis of Gallus gallus

Summary Most of the specific monoamine fluorescence of the fowl neurohypophysis is found in the eminentia mediana and the infundibular stem. The densest accumulation of fluorescent structures is located to the zona externa and the subependymal layer, whereas generally only scattered fluorescence is demonstrable in the fiber layer. The neural lobe tissue is provided with very fine smooth fibers often difficult to distinguish. Spectrofluorimetric determinations have shown that noradrenaline is the major catecholamine in the chick neurohypophysis. From the embryological studies it is evident that the monoamine fluorescence first appears in the subependymal layer, the fiber layer and the neural lobe (after about 15 days of incubation). The zona externa fluorescence is not visible until just before hatching. 10 days after hatching the fluorescence intensity of the chick neurohypophysis is similar to that of the adult. Some comparisons are also made with the appearance of monoamines in the mouse.The authors take great pleasure in expressing their warmest thanks for laboratory facilities and good advice provided by Dr. Bengt Falck at the Institute of Histology, Lund, Sweden.This work was supported by grants from the Swedish Natural Science Research Council (project no. 99-35 and 2180-16), from the United States Public Health Service (NB-06701-02) and from the Swedish Medical Research Council (B-69-14 x -56-05 C).     

Application of a Theory for Circumnutations to Geotropic Movements

From a previously published theory (Israelsson and Johnsson 1967) for circumnutations in Helianthus annuus it is possible to predict the geotropical curvatures of the hypocotyls. This extension of the theory is given in the present paper and some geotropical experiments are performed and discussed. The agreement between the theory and the experiments has been verified in the case of gravitational stimulation during relatively short stimulation periods, in the case of continuous gravitational stimulation, etc. Restrictions in the proposed model are discussed.     

Adrenergic nerves in the pars intermedia of the pituitary in the toad,Bufo arenarum

Summary By means of a highly sensitive and specific histochemical method for the demonstration of certain biogenic monoamines a plexus of nerves containing a primary catecholamine has been demonstrated in the pars intermedia of the toad, Bufo arenarum. These nerves are restricted in distribution to the part of the gland which contains colloid vesicles (stored MSH ?) in the cells. The view is put forward, based on the results of pharmacological and surgical experiments, that the adrenergic nerves inhibit the release of the MSH from the pars intermedia. The origin of the nerves in the brain is unknown, but experiments with lesions have shown that it is not to be found in the nucleus periventricularis arcuatus.The research reported in this document has been sponsored by the Swedish Medical Research Council and by the Swedish Natural Science Research Council.Research fellow of the Commission of Scientific Research of the Province of Buenos Aires, Argentina.