Differential Cellular Enrichment of Gangliosides in the Mouse Cerebellum: Analysis Using Neurological Mutants

Abstract: The cellular distribution of gangliosides in the cerebellum was studied in a series of adult mouse mutants that lose specific populations of neurons. The weaver ( wv ) mutation destroys the vast majority of granule cells, whereas the Purkinje cell degeneration mutation ( pcd ) destroys the vast majority of Purkinje cells. The staggerer ( sg ) and lurcher ( Lc ) mutations, on the other hand, destroy the vast majority of both granule and Purkinje cells. A proliferation of reactive glial cells, which occurs as a consequence of neuronal loss, has been reported in the sg/sg and pcd/pcd mutants, but not in the wv/wv mutant. Compared with the normal (+/+) mice, the concentration (μg/100 mg dry weight) of G_D1a was significantly reduced in those mutants that lost granule cells, but was not reduced in the pcd/pcd mutant. The concentration of G_TIa, on the other hand, was significantly reduced in those mutants that lost Purkinje cells, but was not reduced in the wv/wv mutant. A significant elevation in the concentration of G_D3, which may be related to the proliferation of reactive glial cells, was observed in the pcd/pcd, sglsg , and Lc /+ mutants, but was not observed in the wv/wv mutant. Because these ganglioside abnormalities were confined to the cerebellum, they cannot result from genetic defects in ganglioside metabolism. Instead, these abnormalities result from a differential enrichment of gangliosides in neural membranes. Our findings suggest that G_DT1a is more heavily concentrated in granule cells than Purkinje cells, whereas the opposite appears true for G_Tla. It also appears that GD3 is enriched in reactive glial cells and may play an important role during the morphological transformation of neural membranes.     

Turnaround of Axoplasmic Transport of Selected Particle-Specific Enzymes at an Injury in Control and Diisopropylphosphorofluoridate-Treated Rats

: Reversal of direction (turnaround) of axonal transport of particle specific enzyme activities was studied at a ligature placed on rat sciatic nerve. In the principal experiment, the ligature remained on the nerve in vivo several hours, allowing enzyme activities (acetylcholinesterase, acid phosphatase, and monoamine oxidase) to accumulate immediately proximal to the tie. The nerve was then tied a second time, proximal to the first tie, and incubated in vitro for several more hours. Accumulation of enzyme activities just distal to the second tie was measured. This second accumulation, of activities traveling in the retrograde direction, was shown to be the result of turnaround in several ways. (1) The increase in activity distal to the second tie was equal to the decrease in activity proximal to the first. (2) The increase in enzyme activities distal to the second tie was greatly reduced when the accumulation proximal to the first tie was trapped by placing a third tie between the first and second ties. (3) It was shown that the activity that accumulated distal to the second tie could not have been in retrograde motion at the time of the first tie. (4) Accumulation distal to the second tie was not a function of the length of nerve segment included between the two ties. In contrast to the consistent occurrence of turnaround of orthograde flow, turnaround of retrograde flow could not be demonstrated. Turnaround transport was blocked by incubation in the cold and in the presence of NaCN or vinblastine. The turnaround process operated on all three enzymes studied, suggesting that it operates on lysosomes and mitochondria, as well as on the endoplasmic reticulum-like material bearing acetylcholinesterase. Evidence for the participation of the transport process in the renewal of AChE in the distal portions of the axon was obtained in experiments using diisopropylphosphorofluoridate and cycloheximide.     

De novo biosynthesis of enkephalins and their homologues in the human placenta

Fresh trophoblastic preparations of two human placentae delivered at term were pulse labelled for 30, 120 and 240 min with tritiated L-tyrosine. After deproteinizing and defatting, the peptide extracts were first concentrated through reversible hydrophobic binding on octadecasilyl-silica particles, prior to further resolution by repetetive high-performance liquid chromatography. Four peptides were isolated and purified to radioactive homogeneity, namely Met-enkephalin, Leu-enkephalin, (Arg⁶)-Leu-enkephalin, and (Arg⁶, Arg⁷)-Leu-enkephalin. Their presence and identity were further confirmed by substractive Edman degradation and by radioimmunoassay. No detectable amounts of radioactive Dynorphin could be trapped, however. Under the incubation conditions used, reference tritiated Leu-enkephalin had a biological half-life of circa 9.5 min.     

Different families of double-stranded conformations of DNA as revealed by computer calculations

An algorithm has been developed that permits one to find all possible conformations of the sugar-phosphate backbone for any given disposition of DNA base pairs. For each of the conformations thus obtained, the energy of the helix was calculated by the method of atom-atom potentials. Several isolated regions in the space of the bases′ parameters (Arnott's parameters) have been found for energetically favorable helical structures. Two parameters, the distance of a base pair from the helix axis, D, and the windling angle, τ, allow one to subdivide possible conformations into the families of closely related forms. Two regions (ravines) on the (D, τ) map correspond to the know A and B families. In the B family a continuous transition has been obtained in which the double helix undergoes increasing winding, while the base pairs are moving toward the major (nonglycosidic) groove. Interrelationships between the variables, characterizing the spatial structure of the double helix, D, τ, TL and χ, when going along the bottom of the B ravine, were also obtained. Besides the Known A and B families, several new ones were found to be energetically possible. Among these the strongly underwound helices with the negative D values, as well as the forms with the C4-C5 angle in a trans position, should be mentioned. Biological roles of the different double-stranded conformations, in particular, in protein-nuclei acid interaction are discussed.     

On structure and functioning of ecosystem in a Salmon lake

The seasonal dynamics of the main components of the ecosystem as well as the vertical distribution of their biomass and production were studied in the salmon lake Dalnee at Kamchatka Peninsula. The data received were used for the construction of schemes of energy balance during two successive years: 1970 and 1971, which were different by their thermic regime. Analysis of schemes showed that during a colder season the food chain was shorter and more effective in relation of the use of primary production by zooplankton. Through the detritus pool the ecosystem used from 16% (1971) up to 70% (1970) of energy of primary production. Seasonal succession includes two main phases -autotrophic and heterotrophic. The energy balance of the planktonic community at these two phases is given.     

Coordination of movements of the tube feet and arms of ophiurans during locomotion

The role of the tube feet in locomotion of the ophiuranAmphipholis kochii Lütken is examined. During stepping movements the anterior tube feet attach themselves to the supporting surface and at the end of the step they detach themselves from it. The signal for detachment is deviation of the foot into the posterior (relative to the direction of motion) position. Because of this arrangement of the "stepping" mechanism of the tube feet the task of coordinating their activity with arm movements is greatly simplified. The feet "automatically" attach themselves to the support when the arm which carries them plays the role of motor; conversely, they detach themselves from the support when this arm moves forward. The CNS thus evidently does not participate in foot and arm movement coordination. It simply assigns the general direction of motion to all the feet and coordination takes place "automatically" as a result of the special properties of the "stepping" mechanism of individual feet.Institute of Oceanology, Academy of Sciences of the USSR, Moscow. Institute of Problems of Information Transmission, Academy of Sciences of the USSR, Moscow. Moscow State University. Translated from Neirofiziologiya, Vol. 8, No. 6, pp. 633–639, November–December, 1976.     

Coordination of arm movement during locomotion in ophiurans

During movement of the ophiuranAmphipholis kochii Lutken, any one of its arms can point forward and, consequently, any arm can perform different functions. The arm, when separated from the ophiuran together with the adjacent part of the nerve ring, is capable of complex motor acts, including locomotion. Division of the nerve ring in the ophiuran disturbs coordination of the arms. The results of experiments in which one or more arms were amputated showed that the choice of leading arm and of method of locomotion depends mainly on afferent impulses received from the arms. The results indicate that the neural centers of individual arms possess relative autonomy. Coordinated working of the centers is achieved through their interaction. This interaction ensures the distribution of functions between the arms in accordance with the motor task to be undertaken and coordinates the activity of the arms in time. The dominant role in the distribution of functions between the arms is played by the center of the leading arm, which controls the activity of at least the adjacent centers.Institute of Oceanology, Academy of Sciences of the USSR, Moscow. Institute of Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Moscow State University. Translated from Neriofiziologiya, Vol. 8, No. 5, pp. 529–537, September–October, 1976.     

A new method for determination of elastolytic activity using (14C) labeled elastin and its application to leukocytic elastase.

A new, highly sensitive and specific assay for elastolytic activity is described which employs insoluble elastin randomly labeled with [¹⁴C]. The substrate was prepared by labeling amino groups of the protein in vitro with [¹⁴C] methyl groups by reductive alkylation. The substrate was used to quantitate elastolytic activity from human leukocytes and to compare leukocytic elastase with pancreatic elastase. Purified human leukocytic elastase was approximately one-fourth as active as pancreatic elastase. Similar difference between leukocytic elastase and pancreatic elastase activities was found when the enzymes were tested against succinyl-L-alanyl-L-alanyl-L-alanine-p-nitroanilide, but not when t-BOC-L-alanine-p-nitrophenyl ester was used.