scholarly journals DISTRIBUTION OF LEUCINE-3H DURING AXOPLASMIC TRANSPORT WITHIN REGENERATING NEURONS AS DETERMINED BY ELECTRON-MICROSCOPE RADIOAUTOGRAPHY

1972 ◽  
Vol 52 (3) ◽  
pp. 719-732 ◽  
Author(s):  
Thomas L. Lentz
Keyword(s):  
Electron Microscope ◽  
Short Distance ◽  
Unit Area ◽  
Trophic Factors ◽  
Nerve Terminals ◽  
Sensory Ganglia ◽  
Successive Addition ◽  
Myelinated Nerves ◽  
Almost All ◽  
Silver Grains

The distribution of leucine-3H in neurons was determined by electron-microscope radioautography after infusion of label into the spinal cord or sensory ganglia of regenerating newts. In the nerve cell bodies 3 days after infusion, the highest concentration of label per unit area occurred over the rough-surfaced endoplasmic reticulum. In the large brachial nerves, the silver grains were not distributed uniformly in the axoplasm, indicating that the labeled materials are restricted in their movement to certain regions of the axon. Almost all of the radioautographic grains observed in myelinated nerves could be accounted for by the presence of a uniformly labeled band occupying the area 1500–9000 A inside the axolemma. This region of the axon was rich in microtubules and organelles while the unlabeled central core of the axon contained mainly neurofilaments. This observation supports the hypothesis that microtubules are related to axonal transport. In small, vesicle-filled nerve terminals in the blastema, labeled material was restricted to a thin zone a short distance beneath the plasma membrane while the central region of the terminal was largely unlabeled. The peripheral pattern of labeling in the nerve endings is consistent with successive addition of newly synthesized proteins at the periphery of the growth cone and release of substances such as trophic factors at the nerve terminal.

Quantitative high resolution light and EM autoradiography of fluorophosphate-esterase in developing mouse liver

1978 ◽  
Vol 36 (2) ◽  
pp. 66-67
Author(s):  
C. Budd
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Mouse Liver ◽  
Unit Area ◽  
Irreversible Inhibitor ◽  
Adult Mice ◽  
Quantitative Measurements ◽  
Em Autoradiography ◽  
Old Mice ◽  
Silver Grains

The distribution and concentration of fluorophosphate reactive (FPR) esterases in the liver of developing and adult mice was determined quantitatively from light and electron microscope autoradiograms of liver reacted with 3H - diisopropylfluorophosphate (3H-DFP) an irreversible inhibitor of carboxylesterases.The majority of labeled cells in the liver of 2, 8, 14 and 28- day-old mice and in adult mice (60-180 days old) were hepatocytes, but in the liver from 2-day-old mice and to a lesser extent 8-day-old mice, the autoradiographic silver grains were also concentrated over granulocytes of the intrahepatic hemopoietic population.Quantitative measurements of grain density (developed silver grains/unit area) in light microscope autoradiograms revealed an increase in the predominantly cytoplasmic concentration of FPR esterase sites in hepatocytes from 137o of the adult concentration in 2-day-old mice, to 497, and 78% of the adult concentration in 8- and 14-day-old animals, respectively.

Vacuum System to Minimize the Specimen Contamination of High-Performance EM

1977 ◽  
Vol 35 ◽  
pp. 68-69
Author(s):  
N. Yoshimura ◽  
K. Shirota ◽  
T. Etoh
Keyword(s):  
Electron Microscope ◽  
High Speed ◽  
High Performance ◽  
High Vacuum ◽  
Vacuum System ◽  
Pump System ◽  
Pumping System ◽  
Diffusion Pump ◽  
Almost All ◽  
Cascade Type

One of the most important requirements for a high-performance EM, especially an analytical EM using a fine beam probe, is to prevent specimen contamination by providing a clean high vacuum in the vicinity of the specimen. However, in almost all commercial EMs, the pressure in the vicinity of the specimen under observation is usually more than ten times higher than the pressure measured at the punping line. The EM column inevitably requires the use of greased Viton O-rings for fine movement, and specimens and films need to be exchanged frequently and several attachments may also be exchanged. For these reasons, a high speed pumping system, as well as a clean vacuum system, is now required. A newly developed electron microscope, the JEM-100CX features clean high vacuum in the vicinity of the specimen, realized by the use of a CASCADE type diffusion pump system which has been essentially improved over its predeces- sorD employed on the JEM-100C.

scholarly journals THE ORGANIZATION AND INNERVATION OF THE LUMINESCENT ORGAN IN A FIREFLY, PHOTURIS PENNSYLVANICA (COLEOPTERA)

1963 ◽  
Vol 16 (2) ◽  
pp. 323-359 ◽  
Author(s):  
David S. Smith
Keyword(s):  
Electron Microscope ◽  
Synaptic Vesicles ◽  
Light Emission ◽  
Nerve Endings ◽  
Tracheal Epithelium ◽  
Nerve Terminals ◽  
Tracheal System ◽  
Physiological Evidence ◽  
Effector System

The organization of the luminescent organ of an adult firefly has been studied with the electron microscope, and particular attention has been given to the disposition of nerve terminals within the organ. The cytological structure of the cells of the tracheal system, the peripheral and terminal axons, the photocytes and the cells of the dorsal ("reflecting") layer is described. Previous observations on the peripheral course of nerve branches alongside the tracheal trunks at the level of the dorsal layer and photocyte epithelium have been confirmed, and specialised nerve endings containing axoplasmic components structurally identical with "synaptic vesicles" and "neurosecretory droplets" have been identified, not in association with the surface of the photocytes, but lying between the apposed surfaces of two components of the tracheal epithelium: the tracheal end-cell and the tracheolar cell. These cytological findings are discussed in terms of available biochemical and physiological evidence concerning the mechanism of light emission in the firefly, especially with respect to the possible role of chemical "transmitter" action in triggering a response in a luminescent effector system.

scholarly journals Acetylcholinesterase from the motor nerve terminal accumulates on the synaptic basal lamina of the myofiber.

1991 ◽  
Vol 115 (3) ◽  
pp. 755-764 ◽  
Author(s):  
L Anglister
Keyword(s):  
Basal Lamina ◽  
Ache Activity ◽  
Neuromuscular Junctions ◽  
Motor Nerve ◽  
Synaptic Cleft ◽  
Motor Nerve Terminal ◽  
Nerve Terminals ◽  
Axon Terminals ◽  
Muscle Nerve ◽  
Almost All

Acetylcholinesterase (AChE) in skeletal muscle is concentrated at neuromuscular junctions, where it is found in the synaptic cleft between muscle and nerve, associated with the synaptic portion of the myofiber basal lamina. This raises the question of whether the synaptic enzyme is produced by muscle, nerve, or both. Studies on denervated and regenerating muscles have shown that myofibers can produce synaptic AChE, and that the motor nerve may play an indirect role, inducing myofibers to produce synaptic AChE. The aim of this study was to determine whether some of the AChE which is known to be made and transported by the motor nerve contributes directly to AChE in the synaptic cleft. Frog muscles were surgically damaged in a way that caused degeneration and permanent removal of all myofibers from their basal lamina sheaths. Concomitantly, AChE activity was irreversibly blocked. Motor axons remained intact, and their terminals persisted at almost all the synaptic sites on the basal lamina in the absence of myofibers. 1 mo after the operation, the innervated sheaths were stained for AChE activity. Despite the absence of myofibers, new AChE appeared in an arborized pattern, characteristic of neuromuscular junctions, and its reaction product was concentrated adjacent to the nerve terminals, obscuring synaptic basal lamina. AChE activity did not appear in the absence of nerve terminals. We concluded therefore, that the newly formed AChE at the synaptic sites had been produced by the persisting axon terminals, indicating that the motor nerve is capable of producing some of the synaptic AChE at neuromuscular junctions. The newly formed AChE remained adherent to basal lamina sheaths after degeneration of the terminals, and was solubilized by collagenase, indicating that the AChE provided by nerve had become incorporated into the basal lamina as at normal neuromuscular junctions.

Synthesis and Localization of Sulphated Mucopolysaccharide in Megakaryocytes and Platelets of the Rat, An Analysis by Electron-Microscope Autoradiography

1972 ◽  
Vol 10 (3) ◽  
pp. 705-717
Author(s):  
G. G. MacPHERSON
Keyword(s):  
Electron Microscope ◽  
Golgi Apparatus ◽  
Blood Platelets ◽  
Total Activity ◽  
Electron Microscope Autoradiography ◽  
Dense Granules ◽  
Level Of Activity ◽  
Microscope Autoradiography ◽  
Almost All

Electron-microscope autoradiography has been used to investigate the synthesis and localization of sulphated mucopolysaccharide in megakaryocytes and blood platelets. Following 10-min incubation of bone marrow with 35S-sulpahte in vitro the majority of the activity in megakaryocytes was associated with the Golgi apparatus, but a substantial proportion was associated with other cytoplasmic organelles, suggesting either rapid transport or sulphation of mucopolysaccharide outside the Golgi apparatus. Three hours after the intravenous injection of 35SO4 only a small proportion of the total activity was associated with the Golgi apparatus, most being associated with demarcation membranes and dense granules, while 12 h after injection almost all the activity was associated with demarcation membranes and granules. A rising proportion of activity localized solely on the demarcation membranes suggested that they may possess some activity of their own. Autoradiographs of blood platelets prepared 72 h after the injection of 35SO4 were analysed. It was shown that most of the activity was associated with the α-granules, but there was strong evidence that the platelet membrane possessed a low level of activity.

scholarly journals Differential Uptake of Tritiated Thymidine into Hetero- and Euchromatin in Melanoplus and Secale

1959 ◽  
Vol 6 (3) ◽  
pp. 457-466 ◽  
Author(s):  
A. Lima-de-Faria
Keyword(s):  
Sex Chromosome ◽  
Unit Area ◽  
Tritiated Thymidine ◽  
Early Prophase ◽  
Chromosome Counts ◽  
Heterochromatic Block ◽  
Melanoplus Differentialis ◽  
Photometric Measurements ◽  
Silver Grains ◽  
Prophase Of Meiosis

Grasshoppers of the species Melanoplus differentialis were injected with tritium-labelled thymidine. At intervals thereafter autoradiographic stripping film was applied over Feulgen squashes and sections. In this species during early prophase of meiosis the sex chromosome forms a heterochromatic block large enough to be resolved in tritium autoradiographs. A study of the squash preparations reveals that the sex chromosome is synthesizing DNA at a different period of time from the euchromatic autosomes. Since there is a developmental sequence of spermatocyte cysts along the testicular tubes it is possible from the sections to show that the heterochromatin synthesizes DNA later than does the euchromatin. To find out whether the results obtained in Melanoplus were characteristic of heterochromatin in general, young seedlings of rye were grown in a tritiated thymidine solution and Feulgen squashes were made as for Melanoplus. In rye leaf nuclei there is a large block of heterochromatin constituted by the proximal regions of the chromosomes and a euchromatic one formed by the median and distal regions of the same chromosomes. Here also the heterochromatin synthesizes DNA at a different period of time from the euchromatin. It is concluded that in rye the asynchrony of synthesis occurs within each chromosome. Counts of silver grains over the two types of chromatin in nuclei of Melanoplus and Secale disclosed that the number of grains per unit area was two to three times higher over the heterochromatin. To check the DNA content, Feulgen photometric measurements were made of Melanoplus nuclei at the same stage. The Feulgen and grain counts agree in showing that the heterochromatin contains two to three times more DNA per unit area than the euchromatin.

Mitosis in Mougeotia sp.

1972 ◽  
Vol 50 (9) ◽  
pp. 1811-1816 ◽  
Author(s):  
Carla W. Bech-Hansen ◽  
Larry C. Fowke
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Nuclear Envelope ◽  
Short Distance ◽  
Late Anaphase ◽  
New Information ◽  
Early Anaphase

Combined light and electron microscope observations have provided new information concerning mitosis in Mougeotia. The distribution of microtubules during division suggests that intact wall microtubules moved at preprophase to form the spindle and returned to the cell wall at telophase. During metaphase and early anaphase, chromosomal microtubules were attached to distinct kinetochores; few interzonal microtubules were evident. The subsequent elongation of the spindle at late anaphase was accompanied by the appearance of numerous interzonal microtubules and the loss of the original nuclear envelope. The nucleoli dispersed during prophase and reformed at telophase. The wall septum appeared at prophase but extended only a short distance into the cell by telophase; microtubules were not associated with the developing septum.

scholarly journals Selective binding, uptake, and retrograde transport of tetanus toxin by nerve terminals in the rat iris. An electron microscope study using colloidal gold as a tracer

1978 ◽  
Vol 77 (1) ◽  
pp. 1-13 ◽  
Author(s):  
ME Schwab ◽  
H Thoenen
Keyword(s):  
Electron Microscope ◽  
Tetanus Toxin ◽  
Colloidal Gold ◽  
Smooth Endoplasmic Reticulum ◽  
Specific Binding ◽  
Retrograde Transport ◽  
Nerve Terminals ◽  
Gold Complexes ◽  
Selective Binding ◽  
Gold Particles

A series of specific macromolecules (tetanus toxin, cholera toxin, nerve growth factor [NGF], and several lectins) have been shown to be transported retrogradely with high selectivity from terminals to cell bodies in various types of neurons. Under identical experimental conditions (low protein concentrations injected), most other macromolecules, e.g. horseradish peroxidase (HRP), albumin, ferritin, are not transported in detectable amounts. In the present EM study, we demonstrate selective binding of tetanus toxin to the surface membrane of nerve terminals, followed by uptake and subsequent retorgrade axonal transport. Tetanus toxin or albumin was adsorbed to colloidal gold particles (diam 200 A). The complex was shown to be stable and well suited as an EM tracer. 1-4 h after injection into the anterior eye chamber of adult rats, tetanus toxin-gold particles were found to be selectively associated with membranes of nerve terminals and preterminal axons. Inside terminals and axons, the tracer was localized mainly in smooth endoplasmic reticulum (SER)-like membrane compartments. In contrast, association of albumin-gold complexes with nervous structures was never observed, in spite of extensive uptake into fibroblasts. Electron microscope and biochemical experiments showed selective retrograde transport of tetanus toxin-gold complexes to the superior cervical ganglion. Specific binding to membrane components at nerve terminals and subsequent internalization and retrograde transport may represent an important pathway for macromolecules carrying information from target organs to the perikarya of their innervating neurons.

The Case for Adaptability of the Neuromuscular Junction to Endurance Exercise Training

1998 ◽  
Vol 23 (4) ◽  
pp. 339-360 ◽  
Author(s):  
Robert Panenic ◽  
Phillip F. Gardiner
Keyword(s):  
Electrical Stimulation ◽  
Exercise Training ◽  
Neuromuscular Junction ◽  
Key Words ◽  
Endurance Exercise ◽  
Physiological Function ◽  
Trophic Factors ◽  
Nerve Terminals ◽  
Endurance Exercise Training

Although the adaptability of the neuromuscular junction (NMJ) has been demonstrated using the models of denervation/reinnervation, electrical stimulation, development, aging, and pathological states, relatively little is known about the effects of increased chronic voluntary use on the morphology and physiological function of the NMJ. A review of findings relating to adaptations in the various pre- and postsynaptic components of the NMJ with exercise training is presented. These findings are discussed as they pertain to NMJ function during exercise. Other physiological modulators of the NMJ, such as trophic factors released by nerve terminals and muscles, and circulating substances are discussed in terms of possible roles they may play in training-induced adaptations. Key words: neuromuscular junction, endurance exercise, adaptations, morphology, acetylcholinesterase, physiology, trophic factors

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