scholarly journals Slow transport rates of cytoskeletal proteins change during regeneration of axotomized retinal neurons in adult rats

1990 ◽  
Vol 10 (2) ◽  
pp. 641-648 ◽  
Author(s):  
L McKerracher ◽  
M Vidal-Sanz ◽  
AJ Aguayo
1994 ◽  
Vol 107 (11) ◽  
pp. 3165-3172 ◽  
Author(s):  
I. de Curtis ◽  
G. Gatti

Integrin alpha 6 beta 1 is a laminin receptor involved in adhesion and neurite extension of retinal neurons on laminin. The present study was carried out to identify potential interactions between the alpha 6 beta 1 receptor and cellular proteins that may be involved in integrin signaling and function. For this purpose we have used a biochemical approach based on the solubilization of retinal neurons cultured on laminin with nonionic detergents, followed by centrifugation on sucrose velocity gradients. Analysis of the distribution of the alpha 6 and beta 1 integrin subunits in the gradients showed that they migrate as a large complex after extraction of cells with octylglucoside, but not after Triton X-100 extraction. Cytoskeletal proteins known to localize in adhesion plaques did not comigrate with alpha 6 beta 1 in octylglucoside gradients, while a set of polypeptides whose tyrosine phosphorylation was enhanced by culture on laminin colocalized with alpha 6 beta 1 on the gradients after octylglucoside solubilization. Culture of retinal neurons on bovine serum albumin, a nonadhesive substratum, partially affected the gradient distribution of the receptor after octylglucoside extraction. Furthermore, analysis of the gradient distribution of two alternatively spliced isoforms of the alpha 6 subunit, alpha 6-cytoA and alpha 6-cytoB, showed that the effect of non-adhesion on the sedimentation properties of the two integrin alpha 6 isoforms was more dramatic for alpha 6-cytoB than alpha 6-cytoA. These differences in the sedimentation behaviour indicate distinct biochemical properties of the two alpha 6 isoforms that, together with previous observations on their differential distribution in the developing retina, may reflect functional specificities.


1991 ◽  
Vol 2 (3-4) ◽  
pp. 193-205 ◽  
Author(s):  
Laurie C. Doering

Defined regions (septum, substantia nigra) of the embryonic central nervous system (CNS) were transplanted into the sciatic nerves of young adult rats. Immunocytochemical techniques were used to examine the expression of neurotransmitter related enzymes and neuronal cytoskeletal proteins in the grafts.The origin of the septal grafts was confirmed by immunoreactivity in nenrons to choline acetyltransferase and the β-nerve growth factor receptor (192-IgG). In substantia nigra grafts, neuronal perikarya and processes were identified with an antibody directed against tyrosine hydroxylase. Typical spatial distributions of phosphorylated (Mr200,000) and non phosphorylated (Mr168,000 & 200,000) neurofilaments were observed in the short term (1-2 months) grafts with the monoclonal antibodies RT97 and SMI-32 respectively. Dense dendrite arbors and neuronal cell bodies were immunostained with an antibody that recognizes a high molecular weight microtubule associated protein (MAP2).In the long term (1 year) transplants, prominent cytoskeletal changes in the somata, axons and dendrites of neurons were evident. The cells showed a shift in phosphorylated neurofilament staining from the axon to the soma accompanied by a reduction in axonal immunoreactivity in the adjacent neuropil, Other abnormal features included swollen perikarya, hypertrophied axonal segments and Short segments of kinked axons. Regression of the dendrite trees in the long standing grafts was also apparent when sections were reacted with the MAP2 antibody.These experiments indicate that grafted fetal neurons, isolated in the peripheral nervous system, differentiate and express markers like their counterpartsin situ. After extended time periods under these circumstances, cytoskeletal modifications become apparent in the neurons. These aberrant changes are similar to morphological characteristics associated with aging and neurodegenerative disorders. This experimental paradigm offers a new approach, to study cytoskeletal disturbances in neurons and provides a unique opportunity to examine conditions that may modulate the abnormal changes.


1987 ◽  
Vol 495 (1 Cell and Tiss) ◽  
pp. 1-7 ◽  
Author(s):  
A. J. AGUAYO ◽  
M. VIDAL-SANZ ◽  
M. P. VILLEGAS-PÉREZ ◽  
G. M. BRAY

Author(s):  
Alfredo Feria-Velasco ◽  
Guadalupe Tapia-Arizmendi

The fine structure of the Harderian gland has been described in some animal species (hamster, rabbit, mouse, domestic fowl and albino rats). There are only two reports in the literature dealing on the ultrastructure of rat Harderian gland in adult animals. In one of them the author describes the myoepithelial cells in methacrylate-embbeded tissue, and the other deals with the maturation of the acinar cells and the formation of the secretory droplets. The aim of the present work is to analize the relationships among the acinar cell components and to describe the two types of cells located at the perifery of the acini.


Author(s):  
Beverly L. Giammara ◽  
Jennifer S. Stevenson ◽  
Peggy E. Yates ◽  
Robert H. Gunderson ◽  
Jacob S. Hanker

An 11mm length of sciatic nerve was removed from 10 anesthetized adult rats and replaced by a biodegradable polyester Vicryl™ mesh sleeve which was then injected with the basement membrane gel, Matrigel™. It was noted that leg sensation and movement were much improved after 30 to 45 days and upon sacrifice nerve reconnection was noted in all animals. Epoxy sections of the repaired nerves were compared with those of the excised segments by the use of a variation of the PAS reaction, the PATS reaction, developed in our laboratories for light and electron microscopy. This microwave-accelerated technique employs periodic acid, thiocarbohydrazide and silver methenamine. It stains basement membrane or Type IV collagen brown and type III collagen (reticulin), axons, Schwann cells, endoneurium and perineurium black. Epoxy sections of repaired and excised nerves were also compared by toluidine blue (tb) staining. Comparison of the sections of control and repaired nerves was done by computer-assisted microscopic image analysis using an Olympus CUE-2 Image Analysis System.


Author(s):  
J. R. Kuhn ◽  
M. Poenie

Cell shape and movement are controlled by elements of the cytoskeleton including actin filaments an microtubules. Unfortunately, it is difficult to visualize the cytoskeleton in living cells and hence follow it dynamics. Immunofluorescence and ultrastructural studies of fixed cells while providing clear images of the cytoskeleton, give only a static picture of this dynamic structure. Microinjection of fluorescently Is beled cytoskeletal proteins has proved useful as a way to follow some cytoskeletal events, but long terry studies are generally limited by the bleaching of fluorophores and presence of unassembled monomers.Polarization microscopy has the potential for visualizing the cytoskeleton. Although at present, it ha mainly been used for visualizing the mitotic spindle. Polarization microscopy is attractive in that it pro vides a way to selectively image structures such as cytoskeletal filaments that are birefringent. By combing ing standard polarization microscopy with video enhancement techniques it has been possible to image single filaments. In this case, however, filament intensity depends on the orientation of the polarizer and analyzer with respect to the specimen.


Author(s):  
K.S. Kosik ◽  
L.K. Duffy ◽  
S. Bakalis ◽  
C. Abraham ◽  
D.J. Selkoe

The major structural lesions of the human brain during aging and in Alzheimer disease (AD) are the neurofibrillary tangles (NFT) and the senile (neuritic) plaque. Although these fibrous alterations have been recognized by light microscopists for almost a century, detailed biochemical and morphological analysis of the lesions has been undertaken only recently. Because the intraneuronal deposits in the NFT and the plaque neurites and the extraneuronal amyloid cores of the plaques have a filamentous ultrastructure, the neuronal cytoskeleton has played a prominent role in most pathogenetic hypotheses.The approach of our laboratory toward elucidating the origin of plaques and tangles in AD has been two-fold: the use of analytical protein chemistry to purify and then characterize the pathological fibers comprising the tangles and plaques, and the use of certain monoclonal antibodies to neuronal cytoskeletal proteins that, despite high specificity, cross-react with NFT and thus implicate epitopes of these proteins as constituents of the tangles.


Author(s):  
Tony M. Mosconi ◽  
Min J. Song ◽  
Frank L. Rice

Whiskers or vibrissal follicle-sinus complexes (F-SCs) on the snouts of many mammalian species are structures that have complex, dense sensory innervation. The innervation of F-SCs is remarkably similar in all species with the exception of one site - the inner conical body (ICB). The ICB is an elongated cylindrical structure that encircles the hair shaft near the neck of the follicle. This site has received only cursory attention in ultrastructural studies of the F-SCAdult rats were perfused after the method of Renehan and Munger2. F-SCs were quartered longitudinally and embedded separately in Epon-Araldite. Serial 0.25 μm sections were cut in either the longitudinal or perpendicular plane through the ICB and examined with an AEI EM7 1.2 MV HVEM (Albany, NY) at 1000 KV. Sensory endings were reconstructed from serial micrographs through at least 20 μm in the longitudinal plane and through 10 μm in the perpendicular plane.From two to six small superficial vibrissal nerves converge upon the neck of the F-SC and descend into the ICB. The nerves branch into smaller bundles of myelinated and unmyelinated axons along the dorsal side of the hair shaft.


2019 ◽  
Vol 133 (1) ◽  
pp. 50-58 ◽  
Author(s):  
Nathalie D. Elliott ◽  
Rick Richardson

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