Patterns of Particle Movement in Nerve Fibres In Vitro- an Analysis by Photokymography and Microscopy

1972 ◽  
Vol 11 (3) ◽  
pp. 875-886
Author(s):  
M. BERLINROOD ◽  
S. M. McGEE-RUSSELL ◽  
R. D. ALLEN

Tissue-cultured nerve fibres derived from 3 species of amphibian embryos were composed of bundles of neurites, when viewed by electron microscopy, and contained particles moving in both directions by saltations. Photokymographic analysis of ciné microscopy records taken with Zeiss differential interference optics demonstrated instantaneous particle velocities ranging from 0.15 to 1.8 µm 8-1 (13-155 mm/day) in Rana pipiens cultures; 0.16-0.8 µm s-1 (14-74 mm/day) in Ambystoma tigrinum cultures and 0.6-2.25 µm s-1 (45-194 mm/day) in Xenopus laevis cultures. Measured pathlengths for particles saltations ranged from 4 to 63 µm. No correlations between particle velocity and pathlength were found.

1998 ◽  
Vol 15 (2) ◽  
pp. 319-331 ◽  
Author(s):  
ADRIAN G. PALACIOS ◽  
RANJANA SRIVASTAVA ◽  
TIMOTHY H. GOLDSMITH

Photocurrents from isolated rods of adults and sub-adults of three species of amphibians, Rana pipiens, Ambystoma tigrinum, and Xenopus laevis, were measured with suction pipette electrodes. The intensity for a half-maximal response was 0.91 ± 0.48 photons μm−2 flash−1 (mean ± s.d., 10-ms flashes) for Rana, 0.92 ± 0.44 for Ambystoma, and 6.14 ± 1.33 for Xenopus. The mean number of photoisomerizations at half-saturation was 22 ± 12 for Rana, 50 ± 24 for Ambystoma, and 221 ± 48 for Xenopus. The photocurrent per photoisomerization is several times smaller in Xenopus rods than in the other two species. Spectral sensitivity was measured from 277–737 nm with light polarized both parallel and perpendicular to the planes of the membrane disks. Dichroism fell in the near UV and was absent in the region of absorption by tryptophan and tyrosine. Maximum sensitivity of Rana was at 503.9 ± 2.6 nm (n = 86), and of Ambystoma, 505.8 ± 1.8 nm (n = 24). Animals from these same batches that were sampled by HPLC had no 3-dehydroretinal (retinal2). Xenopus containing about 94% retinal2 and 6% retinal1 had λmax at 519.3 ± 2.7 nm (n = 11). Spectral position of the β-band, estimated by the method of Stavenga et al. (1993), appears to be at longer wavelengths in amphibian photoreceptors than in other vertebrates. Fits of log sensitivity to a normalized-frequency template that tracks the long-wavelength tail of the α-band (Lamb, 1995) show that the rod pigments of Rana and Ambystoma are slightly narrower than those found in the photoreceptors of fish and mammals.


Development ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 283-299
Author(s):  
J. M. W. Slack ◽  
D. Forman

When small explants from early gastrulae of Xenopus laevis are allowed to develop in abuffered salt solution there is a considerable difference between the patterns of differentiation obtained from different dorsoventral levels of the marginal zone. These patternsof differentiation correspond to the fates of the different regions in the course of normal development. They are not altered if several explants of the same type are fused before culture. If a ventral marginal zone explant from Xenopus is cultured in contact with a piece of dorsal marginal zone from the axolotl, it forms structures more dorsal in character than it would in isolation or in normal development. This behaviour is shown only feebly with other regions of the axolotl gastrula. A piece of dorsal marginal zone from Xenopus is not affected in its development by culture in contact with an explant of ventral marginal zone from the axolotl. The dorsalization of ventral marginal zone explants is shown only if there is a large area of direct contact with the dorsal explant and if the pieces remain in contact for a period of 48 h or more. It is proposed that this in vitro interaction is the same as the dorsoventral component of action of the ‘organizer’ graft discovered by Spemann and Mangold.


1983 ◽  
Vol 3 (2) ◽  
pp. 226-230 ◽  
Author(s):  
Lars Edvinsson ◽  
Edward D. Högestätt ◽  
Rolf Uddman ◽  
Ludwig M. Auer

Pial veins, choroid plexus veins, and the cerebri magna vein were investigated with regard to their ultrastructural organization, adrenergic nerve supply, and in vitro reactivity. The vessel walls consisted of a continuous layer of endothelial cells, large amounts of collagenous material, and occasional pericytes. Smooth muscle cells were observed only in a few specimens from the cerebri magna vein. All veins were surrounded by adrenergic nerve fibres. Potassium (124 m M) and noradrenaline (10−5–10−4 M) induced small contractions (0.2–0.5 mN) of isolated veins during in vitro conditions. The magnitude of these responses was less than one-tenth of that obtained in small pial arteries.


Author(s):  
Ann Chidester Van Orden ◽  
John L. Chidester ◽  
Anna C. Fraker ◽  
Pei Sung

The influence of small variations in the composition on the corrosion behavior of Co-Cr-Mo alloys has been studied using scanning electron microscopy (SEM), energy dispersive x-ray analysis (EDX), and electrochemical measurements. SEM and EDX data were correlated with data from in vitro corrosion measurements involving repassivation and also potentiostatic anodic polarization measurements. Specimens studied included the four alloys shown in Table 1. Corrosion tests were conducted in Hanks' physiological saline solution which has a pH of 7.4 and was held at a temperature of 37°C. Specimens were mechanically polished to a surface finish with 0.05 µm A1203, then exposed to the solution and anodically polarized at a rate of 0.006 v/min. All voltages were measured vs. the saturated calomel electrode (s.c.e.).. Specimens had breakdown potentials near 0.47V vs. s.c.e.


Author(s):  
Tai-Te Chao ◽  
John Sullivan ◽  
Awtar Krishan

Maytansine, a novel ansa macrolide (1), has potent anti-tumor and antimitotic activity (2, 3). It blocks cell cycle traverse in mitosis with resultant accumulation of metaphase cells (4). Inhibition of brain tubulin polymerization in vitro by maytansine has also been reported (3). The C-mitotic effect of this drug is similar to that of the well known Vinca- alkaloids, vinblastine and vincristine. This study was carried out to examine the effects of maytansine on the cell cycle traverse and the fine struc- I ture of human lymphoblasts.Log-phase cultures of CCRF-CEM human lymphoblasts were exposed to maytansine concentrations from 10-6 M to 10-10 M for 18 hrs. Aliquots of cells were removed for cell cycle analysis by flow microfluorometry (FMF) (5) and also processed for transmission electron microscopy (TEM). FMF analysis of cells treated with 10-8 M maytansine showed a reduction in the number of G1 cells and a corresponding build-up of cells with G2/M DNA content.


Author(s):  
Godfrey C. Hoskins ◽  
Betty B. Hoskins

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.


Author(s):  
S. E. Miller ◽  
G. B. Hartwig ◽  
R. A. Nielsen ◽  
A. P. Frost ◽  
A. D. Roses

Many genetic diseases can be demonstrated in skin cells cultured in vitro from patients with inborn errors of metabolism. Since myotonic muscular dystrophy (MMD) affects many organs other than muscle, it seems likely that this defect also might be expressed in fibroblasts. Detection of an alteration in cultured skin fibroblasts from patients would provide a valuable tool in the study of the disease as it would present a readily accessible and controllable system for examination. Furthermore, fibroblast expression would allow diagnosis of fetal and presumptomatic cases. An unusual staining pattern of MMD cultured skin fibroblasts as seen by light microscopy, namely, an increase in alcianophilia and metachromasia, has been reported; both these techniques suggest an altered glycosaminoglycan metabolism An altered growth pattern has also been described. One reference on cultured skin fibroblasts from a different dystrophy (Duchenne Muscular Dystrophy) reports increased cytoplasmic inclusions seen by electron microscopy. Also, ultrastructural alterations have been reported in muscle and thalamus biopsies from MMD patients, but no electron microscopical data is available on MMD cultured skin fibroblasts.


Author(s):  
J. H. Hayden

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.


Author(s):  
A. R. Crooker ◽  
M. C. Myers ◽  
T. L. Beard ◽  
E. S. Graham

Cell culture systems have become increasingly popular as a means of screening toxic agents and studying toxic mechanisms of drugs and other chemicals at the cellular and subcellular levels. These in vitro tests can be conducted rapidly in a broad range of relevant mammalian culture systems; a variety of biological and biochemical cytotoxicity endpoints can be examined. The following study utilized human keratinocytes to evaluate the relative cytotoxicities of nitrofurazone (NF) and silver sulfadiazine (SS), the active ingredients of FURACIN(R) Topical Cream and SILVADENE(R) Cream, respectively. These compounds are anti-infectives used in the treatment of burn patients. Cell ultrastructure and elemental composition were utilized as cytotoxicity endpoints.Normal Human Epidermal Keratinocytes (HK) were prepared from the EpiPackTM culture system (Clonetics Corporation, Boulder, CO). For scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cells were seeded on sterile 35 mm Falcon plastic dishes; for elemental microanalysis, cells were plated on polished pyrolytic carbon discs (E. Fullam, Latham, NY) placed in the culture dishes.


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