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Removal of polysialic acid-neural cell adhesion molecule induces aberrant mossy fiber innervation and ectopic synaptogenesis in the hippocampus.

The mossy fiber axons of both the developing and adult dentate gyrus express the highly polysialylated form of neural cell adhesion molecule (NCAM) as they innervate the proximal apical dendrites of pyramidal cells in the CA3 region of the hippocampus. The present study used polysialic acid (PSA)-deficient and NCAM mutant mice to evaluate the role of PSA in mossy fiber development. The results indicate that removal of PSA by either specific enzymatic degradation or mutation of the NCAM-180 isoform that carries PSA in the brain causes an aberrant and persistent innervation of the pyramidal cell layer by mossy fibers, including excessive collateral sprouting and/or defasciculation of these processes, as well as formation of ectopic mossy fiber synaptic boutons. These results are considered in terms of two possible effects of PSA removal: an increase in the number of mossy fibers that can grow into the pyramidal cell layer and an inhibition of process retraction by formation of stable junctions including synapses. As these defects on granule cells in the adult animal and PSA-positive granule cells continue to be produced in the mature brain, the present findings may be relevant to previous studies suggesting that PSA-NCAM function is required for long-term potentiation, long-term depression, and learning behaviors associated with hippocampus.

Animals↗

Ontogeny of cholecystokinin-8-containing neuron system of the rat: an immunohistochemical analysis. I. Forebrain and upper brainstem.

Ontogeny of the cholecystokinin-8 (CCK) neuron system in the forebrain and upper brainstem of the rat was investigated by means of indirect immunofluorescence. CCK cells and fibers first appeared in the developing ventral tegmental area and in the primordium of the medial forebrain bundle, respectively, at gestational day 15 (12-14-mm embryos). From that time, CCK cells appeared in various areas of the forebrain and upper brainstem until birth and reached the maximum content at postnatal day 10. After postnatal day 10, although CCK cells tended to decrease slightly in number, colchicine treatment in the adult rats brought out numerous CCK cells in the same areas. In contrast, although CCK fibers developed only slightly during the fetal period, marked development was seen after birth, particularly between postnatal days 5 and 10. After that time, as the rats grew, CCK fibers continued to increase in number and formed a meshwork of varying density in various areas of the forebrain and upper brainstem.

Animals↗

Detection of biomolecules in the near-infrared spectral region via a fiber-optic immunosensor.

The design, development, and application of a fluorescent fiber-optic immunosensor (FFOI) procedure for the detection of antibody/antigen binding within the near-infrared (NIR) spectral region is reported. The technique was developed through the combined use of fiber-optics, semiconductor laser excitation, fluorescence detection, NIR dye, and immunochemical techniques. The antibody is immobilized on the FFOI's sensing tip and utilized as a recognition component for trace amounts of specific antigen. The FFOI is constructed to utilize antibody sandwich technique. Three individual immunoassays are reported. The first two assays utilize the FFOI and NN382, a commercial NIR dye, for the detection of human immunoglobulin G (IgG). In these assays, goat anti-human IgG antibody (GAHG) is immobilized on the sensitive terminal of the FFOI followed by the exposure of the antibody-coated terminal to human IgG. The probe is then introduced to GAHG labeled with NN382, generating a signal. The third assay utilizes the FFOI for the detection of trace amounts of Legionella pneumophila serogroup 1 (LPS1). In this assay, rabbit anti-LPS1 antibody is immobilized on the sensitive terminal of the FFOI followed by exposure to LPS1. The antigen-coated probe is then treated with monoclonal anti-LPS1 antibody followed by incubation with GAHG labeled with NN382. The assays are optimized to detect the corresponding antigen via the NIR-FFOI. Typical measurements are performed in 10-15 min. A 780-nm semiconductor laser provides the excitation of the immune complex and the resulting emission is detected by a 820-nm silicon photodiode detector. The intensity of the resulting fluorescence is directly proportional to the concentration of the antigen. Solutions of IgG and LPS1 with concentrations as low as 10(-11) M and 0.5 ng/ml, respectively, have been detected with a minimum interference.

Animals↗

Mechanisms of septal lamination in the developing hippocampus analyzed by outgrowth of fibers from septal implants. II. Absence of guidance by degenerative debris.

In previous study (Lewis, E.R., and C. W. Cotman (1980) Brain Res. 96: 307--330; E. R. Lewis and C. W. Cotman, submitted for publication), we have shown that septal tissues grafted to the brain of a neonatal rat will extend fibers into special terminal laminae within the hippocampal formation of the recipient. As a consequence of the implantation surgery, neurons in the developing entorhinal cortex are ablated and it is possible that processes of implanted cells may be guided by degenerating afferent fibers in the host. In order to test this hypothesis, we have characterized the distribution of degeneration argyrophilia that results from the implant surgery and the time course of its removal and examined the growth of septal implants in a host brain devoid of observable degenerative debris. The pattern of degeneration argyrophilia shortly after the implantation surgery did not correspond to the pattern of implant-associated AChE staining in the host hippocampal formation. By 72 hr after surgery, the great majority of argyrophilic debris had been removed. Implants placed in a cavity created 72 hr after surgery demonstrated excellent viability and produced the typical pattern of AChE staining in the hippocampal formation of the host. This suggests that the presence of degenerating host axons is not necessary to guide the ingrowth of axons and establish septohippocampal laminated fields.

Acetylcholinesterase↗

Increase of muscle mitochondrial content with age in murine muscular dystrophy.

Comparison of morphological features of gastrocnemius muscle fibers in normal and dystrophic (dy2J) mice during development was undertaken to determine the time course of increased oxidative capacity in dystrophic fibers. Measurements of mitochondrial volume percent and of Z-line width were made in superficial fast-twitch fibers using electron microscopy and stereological techniques. Dystrophic fibers develop a progressively higher mitochondrial volume percent than normal fibers after 1 month of age. Z-line width is positively correlated with mitochondrial volume percent. The results support the hypothesis that progressive changes in muscle fiber properties result from abnormal neural activity (pseudomyotonia) in dystrophic animals.

Aging↗

Stress fibers in endothelial cells overlying atherosclerotic lesions in rabbit aorta.

Endothelial injury or dysfunction has long been postulated to promote atherogenesis, but structural alterations of endothelium in atherosclerosis have remained obscure. We report the common occurrence of actin-containing stress fibers, stainable by rhodamine-phalloidin, in endothelium overlying atherosclerotic lesions in cholesterol-fed rabbits. Nonlesioned areas in the same aortas showed normal endothelium with minimal development of stress fibers, which was no different from the appearance of endothelium in chow-fed rabbits. Microtubule organization revealed by immunofluorescence appeared normal in all areas. The development of stress fibers may be related to an altered substratum for endothelial attachment. This study provided no evidence to relate stress fiber formation with lesion initiation, but an association with well-developed foam cell lesions was evident.

Animals↗

Development of a new in vitro model of elastic fiber assembly in human pigmented epithelial cells.

OBJECTIVES: We developed an in vitro model of elastic fiber assembly that provides a comparison of the efficiency of different tropoelastin molecules to organize into fibers. DESIGN AND METHODS: Recombinant tropoelastin was added to ARPE-19 cell culture medium. The elastic fiber assembly was evaluated by immunofluorescence staining, the quantitative analysis of cross-linking amino acids, and semi-quantitative analysis of matrix-associated tropoelastin. RESULTS: We confirmed that ARPE-19 cells express fibrillin-containing microfibrils and lysyl oxidase, but they do not express tropoelastin. Immunofluorescence staining showed a dose- and time-dependent increase in the extracellular matrix. The quantity of cross-linking amino acids and matrix-associated tropoelastin also increased together with the matrix-associated elastin. Moreover, the analysis of a radioimmunoprecipitation assay (RIPA) buffer-soluble fraction indicated that tropoelastin interacted with microfibrils and cross-linked elastin was detected as a super molecular complex. CONCLUSION: These observations indicate that this in vitro model is especially useful for the analysis of mechanisms of elastic fiber formation.

Animals↗

Postnatal development and synapse elimination of climbing fiber to Purkinje cell projection in the cerebellum.

Cerebellar climbing fiber (CF) to Purkinje cell (PC) synapses in rodents provides a good model to study mechanisms underlying postnatal development of synaptic functions and elimination of redundant synapses in the central nervous system. At birth, each PC is innervated by multiple CFs. Then, single CF input is selected, matured and strengthened, while surplus CFs are eliminated. By the end of the third postnatal week, most PCs become innervated by single CFs. This up-date article aims to provide an overview of recent studies on the mechanisms of this process.

Afferent Pathways↗

Role of dietary fiber in colon cancer: an overview.

Studies have demonstrated a reduced risk of colon cancer when populations with diets high in total fat switched to a diet high in total fiber and certain whole-grain foods. Case-control studies have shown convincingly the relationship between dietary fiber and colon cancer. Studies in animal models have demonstrated that the inhibitory effects of dietary fiber on the development of colonic neoplasms depend on the nature and source of the fiber; that wheat bran appears to inhibit colon tumor development more consistently than do other dietary sources of fiber, such as oat and corn bran; and that dietary administration of phytic acid, high levels of which are present in wheat bran, inhibits colon carcinogenesis. Human dietary intervention studies have also indicated that the modifying effect of dietary fiber on bacterial enzymes involved in the production of putative colon tumor promoters depends on the type of fiber consumed. Dietary wheat bran, but not oat or corn bran, significantly decreased the levels of several tumor promoters in the colon, independent of stool bulk.

Animals↗

Single-fiber myosin heavy chain polymorphism during postnatal development: modulation by hypothyroidism.

The primary objective of this study was to follow the developmental time course of myosin heavy chain (MHC) isoform transitions in single fibers of the rodent plantaris muscle. Hypothyroidism was used in conjunction with single-fiber analyses to better describe a possible linkage between the neonatal and fast type IIB MHC isoforms during development. In contrast to the general concept that developmental MHC isoform transitions give rise to muscle fibers that express only a single MHC isoform, the single-fiber analyses revealed a very high degree of MHC polymorphism throughout postnatal development. In the adult state, MHC polymorphism was so pervasive that the rodent plantaris muscles contained approximately 12-15 different pools of fibers (i.e., fiber types). The degree of polymorphism observed at the single-fiber level made it difficult to determine specific developmental schemes analogous to those observed previously for the rodent soleus muscle. However, hypothyroidism was useful in that it confirmed a possible link between the developmental regulation of the neonatal and fast type IIB MHC isoforms.

Age Factors↗

Elastic fiber-associated proteins of skin in development and photoaging.

We sought to use antibodies against structural (tropoelastin fibrillin) and nonstructural (decay-accelerating factor [DAF], serum amyloid P -SAP- components of elastic fibers to characterize fiber structure in neonatal skin, normal adult skin and adult skin with solar elastosis from advanced photoaging. We found by immunohistochemistry and by western blotting that DAF, unlike SAP, is present on cutaneous elastic fibers in neonates and young children, suggesting that DAF may play an early, integral role in protecting elastic fibers from destruction by complement. The most superficial portion of oxytalan fibers stained with antibodies against fibrillin and DAF, while anti-tropoelastin stained only the deeper portion of oxytalan fibers. This suggests that deep oxytalan fibers are composed of both elastin and microfibrils, while the most superficial component is composed solely of microfibrillar proteins. Solar elastosis showed increased fibrillin, DAF, tropoelastin and SAP. Thus, solar elastosis is composed of both microfibrillar and elastin proteins.

Adult↗

Ultrastructure of muscle fibers and cells synthesizing DNA in lymph hearts of developing frogs and chick embryos.

The ultrastructure of striated muscle fibers and 3H-thymidine (3HTdr)-labeled cells adjacent to them in the lymph hearts of larvae of Rana temporaria, yearling frogs, and 9- to 13-day-old chick embryos was studied by use of electron-microscopic autoradiography. A comparatively high level of differentiation of lymph-heart muscle fibers was observed not only in yearling frogs but also in larvae. Myosatellites occurred in all stages of development. No mitoses were found in muscle fibers. In 9- to 13-day-old chick embryos the myofibers of lymph hearts were somewhat less differentiated than those of the larvae and yearling frogs. Differentiating sarcomeres were often seen in the sarcoplasm of myofibers of chick embryos. The analysis of the ultrastructure of 3HTdr-incorporating cells shows that 2-4 h after the single 3HTdr administration only mononucleated cells devoid of myofilaments are commonly labeled both in tadpoles and chick embryos. When fixation is postponed by 48-70 h, myonuclei frequently become labeled. Thus, the data obtained support the evidence that proliferation and differentiation processes in the developing muscle tissue of the lymph heart of both species studied are mutually exclusive, similar to the situation in differentiating vertebrate skeletal muscle.

Animals↗

Development and application of microporous hollow fiber protected liquid-phase microextraction via gaseous diffusion to the determination of phenols in water.

A new organic solvent-free microextraction technique termed liquid-gas-liquid microextraction (LGLME) was developed. In this technique, a small amount (6 microl) of aqueous acceptor solution (0.5M NaOH) is introduced into the channel of a 2.65 cm polypropylene hollow fiber. The hollow fiber is then immersed in an aqueous sample donor solution. The aqueous acceptor phase in the channel of the hollow fiber is separated from the sample solution by the hydrophobic microporous hollow fiber wall with air inside its pores. The analytes (phenols) passed through the microporous hollow fiber membrane by gas diffusion and were then trapped by the basic acceptor solution. After extraction, the acceptor solution was withdrawn into a microsyringe and injected into a capillary electrophoresis sample vial for subsequent analysis. Limits of detection of between 0.5 and 10 microg/l for eight phenols could be achieved. The relative standard deviations (n=6) of this technique between 2.7 and 7.6%. The technique also provides good enrichment factors for all the eight analytes.

Chromatography, Gas↗

Immunohistochemical study on the development of serotoninergic neurons in the chick: I. Distribution of cell bodies and fibers in the brain.

The ontogenetic development of the serotoninergic system in the embryonic as well as in posthatching chick brain was studied with an indirect immunohistochemical technique with the aid of a specific antibody to serotonin (5-hydroxytryptamine). By embryonic day 4, rostral and caudal groups of serotonin-immunoreactive cell populations appeared in the mesencephalon and rostral and caudal rhombencephalon. At this stage, the rostral group had a considerable number of labelled cells that sent axons toward more rostral parts of brain, whereas the caudal group consisted of a small number of scattered serotonin-immunoreactive cells. The number of serotonin-positive cells increased with development, such that by embryonic day 8 almost all the serotoninergic cell groups found in the adult chick were already present. Serotoninergic-positive cells appeared in the paraventricular organ of the diencephalon as early as embryonic day 10. Judging from the cytoarchitectural organization of serotonin-immunoreactive cells, all of the serotoninergic cell groups in the chick brain seemed to be fully developed by embryonic day 16. On embryonic day 4, serotonin-immunoreactive fibers were found to enter into the marginal layer of the mesencephalon. Subsequently, serotonin-positive fibers ascended in the marginal layer of the brainstem up to the levels of the diencephalon and to the telencephalon on embryonic day 6 and 8, respectively. Serotonin-positive fibers, which first began to penetrate into the mantle layer on embryonic day 8, reached to the rostral pole of the telencephalon by embryonic day 10. In general, serotonin fibers were found in almost all brain regions by embryonic day 16. However, "terminal formation" in some nuclei did not seem to begin until the late embryonic or posthatching period. These observations indicate that the initial development of serotoninergic cell groups occurs during the first half of the 20th day of the incubation period of the chick. However, a longer time, ranging from early embryonic to posthatching stages, is necessary for the complete development of the serotoninergic projections.

Animals↗

Postnatal development and laminar distribution of noradrenergic fibers in cat visual cortex.

Previous studies have indicated that adrenergic receptors show significant changes either in laminar distribution or in number during the critical period of kitten visual cortex development. In order to further investigate the postnatal development of this neurotransmitter system, especially in relation to the critical period, we used a polyclonal antibody against dopamine-beta-hydroxylase to localize noradrenaline-containing afferents in visual cortex of kittens of various ages from birth to adulthood. In young kittens, less than 2 weeks of age, noradrenergic fibers were sparse, short and randomly oriented, and were concentrated in layer I and in deep cortical layers V and VI. By postnatal day 40, the fibers were present throughout all cortical layers and exhibited higher densities in layers I, II, III, V and VI, with a band of lower staining in layer IV. While tangential fibers predominated in layers I, V and VI, relatively straight radial fibers traversed layers II and III. After postnatal day 40, we did not find major changes in the laminar distribution of adrenergic fibers. This developmental laminar distribution pattern of adrenergic fibers resembles that of the beta-adrenergic receptors that we and others have studied in kitten visual cortex, but differs from that of alpha-adrenergic receptors.

Afferent Pathways↗

Neurotropic influence of mesenchymal limb target tissue on spinal cord neurite growth in vitro.

Peripheral nerve fiber outgrowth from developing spinal cord is proposed to be under the influence of the limb bud target which, at the time of nerve fiber invasion, is in an essentially premuscular, mesenchymal condition. Thus, the true target for elongating spinal nerve fibers in early development is mesenchyme rather than differentiated skeletal muscle. Spinal cord explants derived from stage V larval Rana pipiens were cultured in a defined medium in the presence or absence of mesenchymal limb tissue or limb-conditioned medium (LCM). Analysis of quantified neuritic outgrowth under these conditions demonstrated a dependency on the target tissue for enhanced nerve fiber density and oriented growth. The characteristics of neuritic growth in the presence of limb mesenchyme or LCM changed from the relatively sparse and straight outgrowth of control cords to dense, wavy arborizations. Areas of the cord explants nearest the limb tissue exhibited the greatest increases in nerve fiber density and morphologic complexity. Additionally, an inverse relationship existed between growth enhancement and the cord-to-target distance. Regulation of directed nerve growth in vitro is suggested to result from a diffusible, target-originated growth factor that binds to the attachment substratum as a concentration gradient guidance pathway with implications for mechanisms of in vivo nerve growth.

Animals↗

Fiber and colon cancer.

The human evidence that dietary fiber prevents the development of colon cancer has been reviewed. The correlational studies are consistent with a protective effect in 61.9 per cent of reports. However, these studies are all retrospective and largely uncontrolled. The case-control studies provide evidence of a protective effect in only 48 per cent of reports. It is of additional concern that some human studies have found an association between tumor enhancement and some fiber-containing foods. Whereas this is not sufficient evidence to implicate dietary fiber as a promoter of human colon cancer, it does perhaps argue for a more conservative approach to recommending high-fiber diets as a means of cancer prevention. The animal data show that different sources of dietary fiber produce markedly different effects on colon carcinogenesis. Although some fibers exhibit protective properties, others clearly promote tumor development. The mechanisms behind these opposing actions require further investigation. However, one thing is clear and that is that dietary fibers do modulate the carcinogenic process and as such provide a valuable tool for probing the mechanisms and stages of colon tumor development. Dietary fiber appears to play a major role in the regulation of normal intestinal function and in the maintenance of a healthy intestinal mucosa. Although there is some evidence that a fiber-deficient diet predisposes to colon carcinogenesis, it is still not known whether an increase in fiber consumption will prevent the development of colon cancer. This is further complicated by not knowing what constitutes a normal level of fiber intake. In the interim, physicians should perhaps advise their patients to consume a moderate diet that contains vegetables, fruits, and whole grains. This will provide a varied source of fiber-containing foods and if consumed in sufficient quantity will optimize intestinal transit and bulk according to individual needs. Isolated fiber supplements have not been shown to be effective in colon cancer prevention. Further attempts to be more specific about what type of fiber to recommend seem premature at this time. However, an intake in the range of 20 to 35 gm per day of dietary fiber from foods has recently been recommended by an Expert Panel.

Colonic Neoplasms↗

Forskolin inhibits tension development in detergent-treated cardiac muscle fibers.

In detergent-treated cardiac muscle fibers, forskolin, a potent activator of adenylate cyclases, inhibits tension development elicited with submaximal [Ca2+] and increases incorporation of 32P into troponin-I. A similar reduced tension development has been observed after treatment with cAMP or the catalytic subunit of the cAMP-dependent protein kinase. It is concluded that these fibers still contain much of the enzyme cascade involved in evoking a contractile response to beta-adrenergic stimulation.

1-Methyl-3-isobutylxanthine↗