RESUMEN
Steroid-binding globulins (SBGs) such as sex hormone binding globulin, corticosteroid binding globulin, and vitamin-D binding protein are receiving increasing notice as being actively involved in steroid actions. This paper reviews data of all three of these SBGs, focusing on their presence and possible activity in the brain and nose. We have found all three proteins in the brain in limbic areas such as the paraventricular (PVN) and supraoptic nuclei (SON) as well as other areas of the hypothalamus, hippocampus, and medial preoptic area. There is also evidence that all three are made in the PVN and SON, in conjunction with the neuropeptides oxytocin and vasopressin. The localization of these three SBGs is more variable within areas of the main olfactory area and the vomeronasal organ. However, all three are found in the mucus of these areas, suggesting that one of their functions is to sequester aerosol steroids, such as pheromones, and deliver them to sensory cells and then to deeper sensory areas. In this manuscript, we present multiple models of SBG action including: A) SBG binding to a membrane receptor, B) this SBG receptor being associated with a larger protein complex including cytoplasmic steroid receptors, C) when the SBGs binds to their SBG receptors, second messengers within the cells respond, D) after SBG binding to its receptor, it releases its associated steroid into the membrane's lipid bilayer, from which it gains access into the cell only when bound by an internal protein, E) the SBG, possibly with its bound SBG receptor, is internalized into the cell from which it can gain access to numerous organelles and possibly the cell's nucleus or F) associate with intracellular steroid receptors, G) SBGs produced in target cells are released from those cells upon specific stimulation, and H) according to the Free Steroid Hypothesis steroids released from the extracellular SBG passively diffuse across the plasma membrane of the cell. These models move the area of steroid endocrinology forward by providing important paths of steroid activity within many steroid target cells.
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Encéfalo/metabolismo , Nariz/química , Globulina de Unión a Hormona Sexual/análisis , Globulina de Unión a Hormona Sexual/metabolismo , Animales , HumanosRESUMEN
OBJECTIVE: As prostate-specific antigen (psa) makes prostate cancer (pca) screening more accessible, more men are being identified with conditions that indicate high risk for developing pca, such as elevated psa and high-grade intraepithelial neoplasia (hgpin). In the present study, we assessed psychological well-being and risk perception in individuals with those high-risk conditions. METHODS: A questionnaire consisting of a psychological symptom survey, a trait risk-aversion survey, and a cancer-specific risk perception survey was administered to 168 patients with early-stage localized pca and 69 patients at high risk for pca (n = 16 hgpin, n = 53 psa > 4 ng/mL). Analysis of variance was used to examine differences in psychological well-being and appraisal of risk between the groups. RESULTS: Compared with the pca group, the high-risk group perceived their risk of dying from something other than pca to be significantly lower (p = 0.007). However, pca patients reported significantly more clinically important psychological symptoms. CONCLUSIONS: The identification of prostate conditions that predict progression to cancer might not result in the psychological symptoms commonly experienced by pca patients, but does appear to be related to a distorted perception of the disease's mortal risk. Patients with pca experience reduced psychological well-being, but better understand the risks of pca recurrence and death. Education on the risks and outcomes of pca can help at-risk men to view health assessments with reduced worry.
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There are a lot of publications on the physical fitness of patients with haemophilia (PWH), however, most studies only reflect individual sport-specific motor capacities or focus on a single fitness ability. They involve small patient populations. In this respect principal objective of this study was to compare the physical fitness in all respects and the body composition of young PWH to healthy peers based on the most valid data we could get. Twenty-one German haemophilia treatment centres were visited from 2002 to 2009. PWH between 8 and 25 years were included. They performed a five-stage fitness test covering the sport-specific motor capacities for coordination, measured by one leg stand, strength, aerobic fitness and mobility as well as body composition. The patients' results were compared with age- and gender-specific reference values of healthy subjects. Two hundred and eighty-five PWH (mean age 13.2 ± 4.5 years, 164 PWH with severe disease) were included prospectively in the study. PWH are significantly below the reference values of healthy subjects in the one-leg stand test, the mobility of the lower extremity, the strength ratio of chest and back muscles and the endurance test. In body composition, the back strength and the mobility of the upper extremity PWH are significantly above the reference values. There are no significant differences in abdominal strength. In conclusion we found specific differences in different fitness abilities between PWH and healthy subjects. Knowing this, we are able to work out exercise programmes to compensate the diminished fitness abilities for our PWH.
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Hemofilia A/fisiopatología , Hemofilia B/fisiopatología , Deportes , Tejido Adiposo/diagnóstico por imagen , Adolescente , Adulto , Niño , Alemania , Humanos , Masculino , Fuerza Muscular/fisiología , Resistencia Física/fisiología , Aptitud Física , Equilibrio Postural/fisiología , Estudios Prospectivos , Radiografía , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Central nervous glycogen synthase kinase 3ß (GSK3ß) is implicated in a number of neuropsychiatric diseases, such as bipolar disorder, depression, schizophrenia, fragile X syndrome or anxiety disorder. Many drugs employed to treat these conditions inhibit GSK3ß either directly or indirectly. We studied how conditional knockout of GSK3ß affected structural synaptic plasticity. Deletion of the GSK3ß gene in a subset of cortical and hippocampal neurons in adult mice led to reduced spine density. In vivo imaging revealed that this was caused by a loss of persistent spines, whereas stabilization of newly formed spines was reduced. In electrophysiological recordings, these structural alterations correlated with a considerable drop in the frequency and amplitude of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-dependent miniature excitatory postsynaptic currents. Expression of constitutively active ß-catenin caused reduction in spine density and electrophysiological alterations similar to GSK3ß knockout, suggesting that the effects of GSK3ß knockout were mediated by the accumulation of ß-catenin. In summary, changes of dendritic spines, both in quantity and in morphology, are correlates of experience-dependent synaptic plasticity; thus, these results may help explain the mechanism of action of psychotropic drugs inhibiting GSK3ß.
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Espinas Dendríticas/fisiología , Potenciales Postsinápticos Excitadores/genética , Regulación de la Expresión Génica/genética , Glucógeno Sintasa Quinasa 3/deficiencia , Neuronas/citología , beta Catenina/metabolismo , Animales , Antineoplásicos Hormonales/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Corteza Cerebral/citología , Fármacos actuantes sobre Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Antagonistas del GABA/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/genética , Glucógeno Sintasa Quinasa 3 beta , Hipocampo/citología , Técnicas In Vitro , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Ratones , Ratones Transgénicos , Neuronas/efectos de los fármacos , Técnicas de Placa-Clamp , Picrotoxina/farmacología , Tamoxifeno/farmacologíaRESUMEN
Inclusive paedagogic thinking and acting is a modern and increasingly important topic in school sports. It will affect teachers as well as parents and students. The new international guidelines and national curricula enable new ways of inclusion especially for students with chronic illnesses like haemophilia. Special help from the sport teachers is of vital importance. In our project "fit for life" where we advice children and young adults with haemophilia to find their appropriate sport, we developed a new approach for an optimised inclusion of children with haemophilia into sport lessons. The whole project is running in corporation with the German Sport Teachers Association/Hessen. We analysed and rated the actual curricula of the different school years and looked at the specific needs, risks and necessary abilities for persons with haemophilia. By this means we gathered about 600 typical movements and/or exercises for school sports and developed individual advice and adapted exercise solutions for sport lessons.
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Curriculum , Hemofilia A , Educación del Paciente como Asunto/métodos , Educación del Paciente como Asunto/organización & administración , Servicios de Salud Escolar/organización & administración , Instituciones Académicas/organización & administración , Deportes/educación , Adolescente , Niño , Alemania , Humanos , Enseñanza , Adulto JovenRESUMEN
BACKGROUND: Proof is lacking that pollen count is representative for allergen exposure, also because allergens were found in nonpollen-bearing fractions of ambient air. OBJECTIVE: We monitored simultaneously birch pollen and the major birch pollen allergen Bet v 1 in different size fractions of ambient air from 2004 till 2007 in Munich, Germany. METHODS: Air was sampled with a ChemVol high-volume cascade impactor equipped with stages for particulate matter (PM)>10 microm, 10 microm>PM>2.5 microm, and 2.5 microm>PM>0.12 microm. Allergen was determined with a Bet v 1-specific ELISA. Pollen count was assessed with a Burkard pollen trap. We also measured the development of allergen in pollen during ripening. RESULTS: About 93 +/- 3% of Bet v 1 was found in the PM > 10 microm fraction, the fraction containing birch pollen. We did not measure any Bet v 1 in 2.5 microm > PM > 0.12 microm. Either in Munich no allergen was in this fraction or the allergen was absorbed to diesel soot particles that also deposit in this fraction. Pollen released 115% more Bet v 1 in 2007 than in 2004. Also within 1 year, the release of allergen from the same amount of pollen varied more than 10-fold between different days. This difference was explained by a rapidly increasing expression of Bet v 1 in pollen in the week just before pollination. Depending on the day the pollen is released during ripening, its potency varies. CONCLUSION: In general, pollen count and allergen in ambient air follow the same temporal trends. However, because a 10-fold difference can exist in allergen potency of birch pollen, symptoms might be difficult to correlate with pollen counts, but perhaps better with allergen exposure.
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Aire/análisis , Antígenos de Plantas/análisis , Betula , Monitoreo del Ambiente/métodos , Polen , Antígenos de Plantas/inmunología , Betula/inmunología , Ensayo de Inmunoadsorción Enzimática , Alemania , Polen/inmunología , Rinitis Alérgica Estacional/inmunologíaRESUMEN
Atmospheric circulation leaves few direct traces in the geological record, making reconstructions of this crucial element of the climate system inherently difficult. We produced a regional Mediterranean synthesis of paleo-proxy data from the sea surface to alpine altitudes. This provides a detailed observational context for change in the three-dimensional structure of atmospheric circulation between the Last Glacial Maximum (LGM, approximately 23,000 to 19,000 years ago) and the present. The synthesis reveals evidence for frequent cold polar air incursions, topographically channeled into the northwestern Mediterranean. Anomalously steep vertical temperature gradients in the central Mediterranean imply local convective precipitation. We find the LGM patterns to be analogous, though amplified, to previously reconstructed phases of enhanced meridional winter circulation during the Maunder Minimum (the Little Ice Age).
RESUMEN
Stretch of nerve has been reported to decrease the amplitude of the compound action potential (CAP) with a complete block appearing in approximately 30 minutes. But for the most part, those experiments were carried out in vivo, and it is generally accepted that the failure of responses was due to a closure of vessels supplying the nerve with a resulting ischemia and anoxia. These studies were undertaken to determine if stretch of nerve has effects that are independent of interference with its vascular supply. In the studies, lengths of rat sciatic and dog peroneal nerves were removed and placed in a chamber supplied with oxygen in which their CAPs were continuously elicited and recorded. This in vitro preparation obviated interference with the nerve's metabolism on stretching. We have previously shown that the form change termed 'beading,' appearing within 10 seconds and reversing as quickly on relaxation, can be elicited with tensions of only several grams. We wished to determine if stretch adequate to produce beading could alter CAPs with the same rapidity. Tensions below 2 g had little effect. On applying tensions of 10-100 g, levels well above those needed to bead the fibers, both increases and decreases of CAP amplitude were seen. The changes occurred within 10 seconds of stretch application, the time at which beading arises with stretch. Although the decreases of CAP amplitudes could be accounted for by beading, the degree of CAP change did not correspond to the amount of tension applied. We hypothesize that the constrictions in the beaded fibers increase axial resistivity and diminish local currents so as to block conduction. The lack of an increasing degree of decreased CAP amplitude with increases in tension is ascribed to the inhibition of elongation offered by the collagen fibrils present in nerve. Collagenase applied to nerves allowed a further increase in length, producing a 'hyperbeading,' showing much longer lengths of beading constrictions on stretch. This would further increase axial resistance and is taken to account for the greater decreases of CAP amplitudes seen following collagenase treatment. To account for those cases where increases of CAP amplitude were seen on stretch, we hypothesize that stretch can also cause an increase in the excitability of the nodes. The outcome of stretch in any given nerve would be the resultant of two opposing actions; beading of the internodes causes a decrease of local currents leading to block of CAPs, while an increased excitability of the nodes acts to augment the responses.
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Nervio Ciático/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Colagenasas/farmacología , Perros , Masculino , Fibras Nerviosas/efectos de los fármacos , Fibras Nerviosas/fisiología , Fibras Nerviosas/ultraestructura , Nervio Peroneo/efectos de los fármacos , Nervio Peroneo/fisiología , Nervio Peroneo/ultraestructura , Estimulación Física , Ratas , Ratas Sprague-Dawley , Valores de Referencia , Nervio Ciático/efectos de los fármacos , Nervio Ciático/ultraestructuraRESUMEN
To account for the beading of myelinated fibers, and axons of unmyelinated nerve fibers as well of neurites of cultured dorsal root ganglia caused by mild stretching, a model is presented. In this model, membrane tension and hydrostatic pressure are the basic factors responsible for axonal constriction, which causes the movement of axonal fluid from the constricted regions into the adjoining axon, there giving rise to the beading expansions. Beading ranges from a mild undulation, with the smallest degree of stretch, to more globular expansions and narrow intervening constrictions as stretch is increased: the degree of constriction is physically limited by the compaction of the cytoskeleton within the axons. The model is a general one, encompassing the possibility that the membrane skeleton, composed mainly of spectrin and actin associated with the inner face of the axolemma, could be involved in bringing about the constrictions and beading.
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Fibras Nerviosas/fisiología , Fibras Nerviosas/ultraestructura , Animales , Fenómenos Biofísicos , Biofisica , Gatos , Substitución por Congelación , Ganglios Espinales/fisiología , Ganglios Espinales/ultraestructura , Técnicas In Vitro , Microscopía Electrónica , Modelos Neurológicos , Fibras Nerviosas Mielínicas/fisiología , Fibras Nerviosas Mielínicas/ultraestructura , Ratas , Nervio Ciático/fisiología , Nervio Ciático/ultraestructura , Estrés MecánicoRESUMEN
Nerve fibers which appear beaded (varicose, spindle-shaped, etc.) are often considered the result of pathology, or a preparation artifact. However, beading can be promptly elicited in fresh normal nerve by a mild stretch and revealed by fast-freezing and freeze-substitution, or by aldehyde fixating at a temperature near 0 degree C (cold-fixation). The key change in beading are the constrictions, wherein the axon is much reduced in diameter. Axoplasmic fluid and soluble components are shifted from the constrictions into the expansions leaving behind compacted microtubules and neurofilaments. Labeled cytoskeletal proteins carried down by slow axonal transport are seen to move with the soluble components and not to have been incorporated into and remain with, the cytoskeletal organelles on beading the fibers. Lipids and other components of the myelin sheath are also shifted from the constrictions into the expansions, with preservation of its fine structure and thickness. Additionally, myelin intrusions into the axons are produced and a localized bulging into the axon termed "leafing". The beading constrictions do not arise from the myelin sheath: beading occurs in the axons of unmyelinated fibers. It does not depend on the axonal cytoskeleton: exposure of nerves in vitro to beta, beta'-iminodipropionitrile (IDPN) disaggregates the cytoskeletal organelles and even augments beading. The hypothesis advanced was that the beading constrictions are due to the membrane skeleton; the subaxolemmal network comprised of spectrin/fodrin, actin, ankyrin, integrins and other transmembrane proteins. The mechanism can be activated directly by neurotoxins, metabolic changes, and by an interruption of axoplasmic transport producing Wallerian degeneration.
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Fibras Nerviosas/fisiología , Fibras Nerviosas/ultraestructura , Animales , Axones/ultraestructura , Citoesqueleto/ultraestructura , Humanos , Modelos Neurológicos , Vaina de Mielina/ultraestructura , Orgánulos/ultraestructura , Estimulación FísicaRESUMEN
Myelinated nerve fibres become beaded when nerves are subjected to a mild stretch; the beading is seen as varicosities, a series of alternating constrictions and enlargements, when using freeze-substitution or cold-fixation to hold this labile form change in place during fixation. One possibility for how this form change comes about is that the myelin sheath or its Schwann cell initiates beading. We now report, however, that a similar beading is seen in the axons of unmyelinated fibres. In electron micrographs, longitudinal sections of axons show the series of constrictions and expansions typical of beading. In cross-sections, axons with unusually small diameter, corresponding to the constrictions, are seen to contain closely packed microtubules and neurofilaments while neighbouring swollen axons with widely dispersed microtubules correspond to the beading expansions. Another possibility for the form change is that the cytoskeleton is responsible for beading. We discovered that direct exposure of nerves to beta, beta'-iminodipropionitrile in vitro for 1-6 h causes both axonal microtubules and neurofilaments to become degraded and replaced by an amorphous residue. Nevertheless, beta,beta'-iminodipropionitrile-treated nerves show constrictions in myelinated fibres when stretched. An even greater degree of beading with narrower and longer constrictions appears in some fibres, with the expanded regions having oblate ends giving the appearance of a string of sausages. In cross-sections taken through the constrictions, a greater than usual reduction of axonal area was seen, this was due to the loss of cytoskeletal organelles which would act to limit the degree of constriction. With longer exposure to beta, beta'-iminodipropinitrile more fibres show complete degeneration of the cytoskeleton and form ovoids typical of Wallerian degeneration. Unmyelinated axons of beta, beta'-iminodipropionitrile-treated nerves which showed degeneration of their cytoskeleton with its replacement by amorphous material still demonstrated beading. As neither the myelin sheath nor the intact cytoskeleton within the axon is necessary for beading, by exclusion, we consider beading constrictions to be initiated at the level of the axolemma. In our hypothesis the membrane skeleton is responsible; namely, the spectrin, actin and other molecular species lining the inside of the axolemma and binding to transmembrane proteins. The membrane skeleton may be activated by stretch via transmembrane proteins (e.g. beta 1-integrins). The membrane skeleton mechanism may also be directly engaged in the production of Wallerian degeneration or be induced by neurotoxic agents.
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Axones/fisiología , Citoesqueleto/fisiología , Fibras Nerviosas/fisiología , Nitrilos/farmacología , Animales , Axones/efectos de los fármacos , Citoesqueleto/efectos de los fármacos , Masculino , Microscopía Electrónica , Modelos Neurológicos , Vaina de Mielina/fisiología , Fibras Nerviosas/efectos de los fármacos , Fibras Nerviosas/ultraestructura , Ratas , Ratas Sprague-DawleyRESUMEN
When the nerves are lightly stretched and fixed by freeze-substitution, their fibers show the form-change termed "beading" which consists of a series of undulating constrictions and swellings in the internodes. This form change has not ordinarily been seen in chemically fixed nerves, or when it has, it has been ascribed to a pathological change or an artifact. We now report that beading is also retained in normal nerves when, following a light maintained stretch, they are fixed with aldehydes at a temperature close to 0 degrees C. The degree of beading in single fibers teased from the aldehyde fixed nerves was graded and found to be maximal at 0 degrees C, falling off with increased temperature until, at temperatures above 16 degrees C, most fibers showed no beading or a very mild beading. The fibers of nerves cold-fixed at 0 degrees C displayed the characteristics as freeze-substituted fibers, but with a somewhat smaller number of maximally beaded fibers and an 18% reduction in microtubule numbers in the axons. Desheathing or slitting the sheaths of the nerves before cold-fixation increased the probability of retaining beading. Exposure of stretched nerves to the aldehyde fixative at room temperatures for times as short as 3-5 min before they were cold-fixed showed a diminished degree of beading, indicating that aldehydes can have a deleterious effect on the beading mechanism which we hypothesize to be present in the fiber. This action is distinct from the general cross-linking action of aldehydes.
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Fibras Nerviosas Mielínicas/ultraestructura , Animales , Artefactos , Reactivos de Enlaces Cruzados/farmacología , Citoesqueleto/ultraestructura , Fijadores/farmacología , Formaldehído/farmacología , Substitución por Congelación , Glutaral/farmacología , Masculino , Vaina de Mielina/efectos de los fármacos , Vaina de Mielina/ultraestructura , Fibras Nerviosas Mielínicas/efectos de los fármacos , Polímeros/farmacología , Ratas , Ratas Sprague-Dawley , Nervio Ciático/ultraestructura , Estrés Mecánico , Fijación del Tejido/métodosRESUMEN
The bands of Fontana, appearing as spirals or irregular light and dark strips crossing the surface of unstretched nerves, are due to the wavy disposition of nerve fibers within the epineural-perineural sheaths. A mean tension of 2.7 +/- 0.23 (S.E.M.) g applied to segments of rat tibial nerves straightens the fibers and unbands the nerves causing them to lengthen by 9.35 +/- 0.89%. The nerves cold-fixed in situ at that point showed the myelinated fibers to be beaded. On relaxation the nerves rebanded and the fibers were no longer beaded. The tension at which unbanding occurred was better determined when the epineural-perineural sheaths were slit longitudinally. Under these conditions, unbanding occurred at a mean tension of 0.59 +/- 0.08 g and the nerves lengthened by 8.56 +/- 0.58%. The lengthening was not statistically different from that seen in sheathed nerves. In preparations with the epineural-perineural sheaths removed, banding was lost with tensions of 0.20 +/- 0.03 g and the nerves lengthened by 12.1 +/- 1.04%. The tensions needed were significantly lower than that for the sheathed and slit-sheath nerve groups. When cold-fixed, when banding was lost, the fibers were seen to be beaded. Banding of the desheathed nerves returned on relaxation of the nerves. However, after tensions of 8 g they showed plasticity in which the ends of the nerves needed to be pushed together to initiate rebanding in comparison to sheathed or sheath-slit nerves which rebanded spontaneously following relaxation after even higher tensions of 40 g. At the highest tensions the nerves remained extended and could not be forcibly rebanded.(ABSTRACT TRUNCATED AT 250 WORDS)
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Fibras Nerviosas Mielínicas/ultraestructura , Manejo de Especímenes/métodos , Potenciales de Acción , Animales , Frío , Colagenasas/farmacología , Masculino , Vaina de Mielina/efectos de los fármacos , Vaina de Mielina/ultraestructura , Fibras Nerviosas Mielínicas/efectos de los fármacos , Fibras Nerviosas Mielínicas/fisiología , Ratas , Ratas Sprague-Dawley , Estrés Mecánico , Nervio Tibial/ultraestructura , Fijación del TejidoRESUMEN
The amounts of apolipoprotein (apo) E and A1 released into the culture medium were examined in the regenerating nerves distal to a crush site following chronic alcohol feeding. Cultured minced segments of regenerating nerves taken from rats fed an alcohol-containing liquid diet for 5 weeks released only 50% of apoE but nearly 200% of apoA1 when compared with rats pair-fed with a control diet. The extent of decrease in medium apoE corresponded to the decrease of apoE mRNA in the nerve. Thus, chronic alcohol ingestion affects apoE synthesis of regenerating nerves by changing its mRNA level. On the other hand, apoA1 mRNA remained undetectable in regenerating and intact nerves whether the rats were fed alcohol or not. Furthermore, the amount of apoA1 released by the regenerating nerve into the culture medium was not significantly larger than that present in the nerve tissue prior to incubation. Therefore, it is most likely that apoA1 released by the injured nerve originated from the bloodstream and the increase in apoA1 content seen in the crushed nerve of alcohol-fed rats is due to an enhanced permeability of the nerve-blood barrier. Since the burst of apolipoproteins in the injured nerve is likely to play a role in nerve regeneration, the perturbation of apolipoprotein contents in regenerating nerves by chronic alcohol consumption may contribute to the pathogenesis of alcoholic neuropathy.
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Alcoholismo/fisiopatología , Apolipoproteínas/metabolismo , Regeneración Nerviosa/efectos de los fármacos , Nervio Ciático/efectos de los fármacos , Animales , Apolipoproteína A-I/metabolismo , Apolipoproteínas E/metabolismo , Transporte Axonal/efectos de los fármacos , Transporte Axonal/fisiología , Northern Blotting , Técnicas de Cultivo , Masculino , Regeneración Nerviosa/fisiología , ARN/metabolismo , Ratas , Ratas Wistar , Nervio Ciático/fisiopatologíaRESUMEN
Small intrusions form in the internodes in or near the constrictions of beaded fibers prepared by fast-freezing and freeze-substituting mildly stretched nerves in the cat and rat. They appear as inwardly directed folds of the inner lamellae of the myelin sheath, or regularly formed spheres composed of lamellae with major dense and interperiod lines like those of the myelin sheath. A splitting of the lamellae and separation of the major dense lines may occur with an accumulation of Schwann cell cytoplasm between them, the result of an influx of cytoplasmic fluid from nearby constrictions. Longitudinally oriented microtubules have been observed in the intrusions, in the adaxonal Schwann cell cytoplasm, and in the innermost lamellae of the myelin sheath. The paranodes contain a number of larger intrusions in the form of spurs and globules along with shelve-like folds of the myelin sheath oriented in the longitudinal direction. Axoplasmic fluid driven from the constrictions during beading can enter the paranodes to smooth out their folds leaving the globular and spur-shaped myelin intrusions in isolation. Their wall thickness, measured from the central opening to the surface of the intrusion, is the same as that of the myelin sheath or, in some cases, double, the result of the folding of a spur-like intrusion upon itself. Intrusions unconnected to the sheath are seen in unbeaded fibers with regular, compact lamellae surrounded by axolemma. Others lack a covering axolemma and consist of variably disorganized and irregularly shaped lamellae suggesting that they are undergoing fragmentation and dissolution within the axon. The hypothesis is advanced that the intrusions in the internodes arise from an excess of lipid and other myelin components when the diameter of the sheath is reduced in the beading constrictions. In the paranodes, excess myelin components moved into these regions form the shelf-like folds which may fuse to form intrusions. These, separated from the myelin sheath, undergo fragmentation and dissolution and are carried by retrograde transport to the cell bodies where their constituent components can be reutilized.
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Vaina de Mielina/ultraestructura , Fibras Nerviosas/ultraestructura , Neuronas/ultraestructura , Animales , Transporte Axonal , Gatos , Congelación , Glutaral , Microscopía Electrónica , Microtúbulos/ultraestructura , Coloración y EtiquetadoRESUMEN
To account for the transport in nerve fibers of tubulin and neurofilament proteins in slow component a, the Structural Hypothesis holds that these proteins are assembled into microtubules and neurofilaments in the cell bodies and the cytoskeletal organelles then moved down in the fibers as part of an interconnected matrix at a uniform rate of about 1 mm/day. The Unitary Hypothesis, on the other hand, considers these proteins to be carried down within the fibers as soluble components or as freely movable small polymers or subunits turning over locally in the stationary cytoskeleton. To differentiate between the two hypotheses, cat L7 dorsal roots were taken at times from 7 to 25 days after their L7 dorsal root ganglia were injected with [3H]leucine to assess the labeling of the cytoskeleton by the use of beading and autoradiography. Beading was induced by a mild stretch and after fast-freezing and freeze-substitution of the roots for histological preparation, the beads were seen in the fibers as a series of expanded regions alternating with constrictions. In the constrictions the cytoskeleton was compacted into an area as small as 5% that of the normal axon, with the axoplasmic fluid and displaceable (freely movable) components squeezed from the constrictions into the adjoining expansions. Roots taken after 7 and 14 days, times consistent with slow component a downflow, were assessed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and their content of tubulin and neurofilament proteins shown to constitute 40-50% of all the labeled proteins present. In autoradiographs of dorsal roots taken at those times, numerous grains due to radioactivity were located over the non-constricted regions of the fibers. Few or no grains were present over the constrictions after 7 days. The findings are in accord with the labeled tubulins and neurofilament proteins being present in soluble form in the fibers and expressed from the constrictions into the expansions of the beaded fibers. In contrast, a number of fibers in roots taken at 14-20 days after injection showed somewhat higher grain densities over the constrictions, and more so after 25 days, indicating uptake of labeled subunits into the cytoskeletal organelles at later times. The results are consistent with the downflow of tubulin and neurofilament proteins as soluble components which drop off in the axon to turn over locally in their respective cytoskeletal organelles.
Asunto(s)
Transporte Axonal , Proteínas del Citoesqueleto/metabolismo , Proteínas de Filamentos Intermediarios/metabolismo , Raíces Nerviosas Espinales/fisiología , Tubulina (Proteína)/metabolismo , Animales , Autorradiografía , Gatos , Leucina/metabolismo , Proteínas de Neurofilamentos , Raíces Nerviosas Espinales/citología , Raíces Nerviosas Espinales/metabolismoRESUMEN
Fast transport of labeled proteins in vertebrate nerve and the particle movement seen in the axoplasm of giant fibers with video microscopy share basic properties. They are both dependent on oxidative metabolism providing ATP to drive transport with the microtubules acting as the "rails" for transport. Glucose is the usual metabolite but when glucose is deleted from an in vitro medium some other endogenous metabolite(s) present in vertebrate nerve can temporarily provide ATP to carry on transport for several hours. The transport filament hypothesis was proposed to account for fast transport in vertebrate nerve. Carriers were pictured to which the various components transported are bound and these moved along the microtubules with the needed energy supplied by ATP utilized by a calmodulin activated Ca-Mg ATPase. Analogous models based on video visualization of particle movement have been proposed. While these models can in general account for fast transport, we require a better understanding of the wide range of slower and intermediate rates of transport seen in vertebrate nerve. The large component of cytoskeletal proteins moved down as slow component a (SCa) at a rate close to 1 mm/d has been accounted for by two different models. In the Structural Hypothesis the microtubules and neurofilament cytoskeletal organelles are considered to be assembled in the cell bodies and then moved down within the axon as an assembled matrix. In the Unitary Hypothesis the cytoskeletal proteins are moved down as soluble proteins on carriers as in the case of fast transport, but these slow transported proteins are more rapidly dropped off in the axon for turnover in the stationary cytoskeleton. Other slow and intermediate rates can similarly be accounted for on the basis of this one mechanism. Experimental evidence in support of the Unitary Hypothesis is presented based on the use of the beading phenomenon to partition the assembled cytoskeleton from the soluble components. Additional evidence is given for a distal assembly of the cytoskeleton in regenerating fibers which further supports Unitary Hypothesis. The Unitary Hypothesis not only best fits with the known properties of transport, it offers new approaches to an understanding of the action of neurotoxins and the genesis of neuropathological changes.