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Historical perspective as a guide for identifying and developing applicable methods for dietary fiber.

A review is presented describing the nature and evolving definition of dietary fiber. The historical development of the current definition is discussed as are the efforts to develop analytical methods to support food labeling regulations. Also considered are the characterization and quantitation of resistance starch, a dietary starch that does not digest in the small intestine, behaves like dietary fiber and therefore may have potential as a health-related ingredient in foods. The current status of AOAC methodology is discussed along with the possibility of updating the definition of dietary fiber. The potential impacts of changing the dietary fiber definition on analytical issues and on food composition databases are also considered.

Dietary Carbohydrates↗

Rapid onset of neuronal death induced by blockade of either axoplasmic transport or action potentials in afferent fibers during brain development.

We have investigated how neurons in the optic tecta of embryonic day 16 chick embryos depend for survival on their afferents from the retina. To distinguish between activity-mediated effects and other, "trophic," ones, we compared the effects on the tectal neurons of blocking intraocular axoplasmic transport (with colchicine) or action potentials (by means of TTX). Both interventions rapidly induced the appearance of dying (pyknotic) neurons in the tectum, with major increases in their number occurring within 13 hr post-colchicine and within 9 hr post-TTX. Following both drugs, the dying neurons were morphologically similar, and in both cases the cell death depended on protein synthesis. However, the effects of colchicine and of TTX could be dissociated, since the most superficial tectal neurons became pyknotic only in response to colchicine, and, with a sufficiently short survival time (9 hr), the deep cells of the stratum griseum centrale became pyknotic only in response to TTX. We hence argue that the survival of the tectal neurons depends on their ongoing maintenance by substances released from retinotectal axon terminals, the release being activity dependent in the case of the deep neurons but independent of activity in the case of the superficial ones.

Action Potentials↗

Distribution of conduction system fibers in the developing and adult rabbit heart revealed by an antineurofilament antibody.

Using an immunological approach, we demonstrated previously that a neurofilament-like protein is expressed in rabbit heart conduction tissue myocytes, and we proposed that these specialized cardiac muscle cells are of neuroectodermal origin. In the present study, we used the expression of the neurofilament-like protein as a marker for identifying conduction tissue cells and studying their distribution in the developing heart. In 11-day-old rabbit embryos, myocytes expressing the neurofilament-like protein were localized at the atrioventricular and the sinoatrial junctions and had a ring-like distribution. At embryonic day 12, reactive myocytes were found also in the subendocardial layer of the dorsal ventricular wall, in continuity with labeled myocytes at the atrioventricular junction. Examination of older embryos and of neonatal and adult hearts revealed that the expression of the neurofilament-like protein was not restricted to myocytes of conduction tissue regions, but it was also detectable in myocytes of the sinoatrial ring bundle, in scattered myocytes localized in the left sinal horn wall, and in the right atrium in proximity to atrioventricular sulcus tissue. Thus, using an intracellular marker, we show that precursors of adult atrial conduction tissue are distributed at the sinoatrial and atrioventricular junctions; at variance, ventricular conduction tissue precursors do not have a ring-like distribution but are localized in the subendocardial layer, in continuity with the atrioventricular junctional myocytes.

Aging↗

Cytoplasmic stress fibers in the developing heart.

Rhodamine-conjugated phalloidin staining was used to study the distribution of filamentous actin in the developing heart of embryonic chicks and rats during the morphogenetic period of cardiac septation. In the chick, intense fluorescence indicative of abundant filamentous actin was observed along the myocardium and in the mesenchymal condensations that formed within the aorticopulmonary septum at day 5. Such cellular condensations and concentration of filamentous actin were not seen in the atrioventricular cushions nor in the preseptation outflow tract. Similar results were found in the 14-day rat embryo. In electron micrographs, microfilament bundles with irregular dense bodies were seen in elongated mesenchymal cells between the valve sites of both species. Cell-cell contacts were observed between such elongated cells and myocyte processes protruding from the nearby myocardial sheath. These histochemical and ultrastructural observations suggest that such mesenchymal condensations serve a specialized mechanical tensile role during embryonic septation of cardiac outflow channels.

Actins↗

Histochemical evidence of altered development of cholinergic fibers in the rat dentate gyrus following lesions. II. Effects of partial entorhinal and simultaneous multiple lesions.

It has been concluded previously that the septohippocampal fibers which project to the rat dentate gyrus extend or branch in the denervated area of the molecular layer following a complete ipsilateral entorhinal lesion. The septohippocampal fibers thus appear to replace some of the perforant fibers which degenerate as a result of the lesion. The reactive fibers eventually become localized to a much smaller and more superficial area after lesions of immature rats than after lesions made in adulthood. To determine whether this difference in the response results from a selective reaction to loss of the lateral perforant path in the immature rat, various portions of the entorhinal cortex were removed at the age of 11 days, and the cholinergic septohippocampal fibers were visualized by acetylcholinesterase histochemistry. An alternative possibility, that the difference between immature and adult rats is attributable to an interaction with other reactive afferents, was tested by removing other sources of input (the contralateral entorhinal cortex, contralateral hippocampal formation or both) along with the ipsilateral entorhinal cortex at the age of 11 days and then demonstrating the septohippocampal fibers histochemically. Lesions of the lateral part of the ipsilateral entorhinal cortex (source of the lateral perforant path) at 11 days of age evoked a septohippocampal reaction along the outer edge of the molecular layer, where the lateral perforant path fibers normally terminate. This result matched that produced by a complete entorhinal lesion. Lesions of the medial entorhinal cortex evoked no obvious reaction. In contrast, the septohippocampal fibers in adult rats proliferated in the denervated area of the molecular layer after lesions of either part of the entorhinal cortex. Combining lesions of other sources of innervation to the dentate gyrus with an ipsilateral entorhinal lesion at 11 days of age did not alter the response of septohippocampal fibers, as determined histochemically. Neither did the septohippocampal fibers react to removal of commissural afferents alone. The response at any age was unaffected by prior or subsequent removal of the contralateral entorhinal cortex. These results indicate that in immature rats the septohippocampal fibers respond only to loss of the lateral perforant path, but these same fibers can later react to loss of any part of the perforant path. They are regarded as support for the hypothesis that the reactive septohippocampal fibers preferentially interact with dendritic growth cones. Our results do not support explanations based on a hypothetical attraction between septohippocampal and crossed perforant path fibers (which react in the same area) or on competition with commissural fibers (which reinnervate an adjacent area). We suggest further that proximity to the degenerating elements does not in itself determine the pattern of reinnervation after lesions of the central nervous system.

Acetylcholinesterase↗

LHRH neurons migrate into the trigeminal nerve when the developing olfactory nerve fibers are physically interrupted in chick embryos.

Most LHRH neurons actively migrate from the olfactory epithelium to the forebrain during embryonic days (ED) 3.5-8. When a small piece of the membrane filter was placed on the central course of the olfactory nerve in ED 3.5-5 chick embryos, LHRH neurons deviated from their regular migratory course at ED 6.5-7.5 to follow a route along the PSA-NCAM-positive medial and lateral nasal branches of the ophthalmic nerve of the trigeminal nerve. The olfactory nerve fibers which were specifically immunoreactive for somatostatin also deviated into the ophthalmic nerve. Enzymatic removal of PSA using endoneuraminidase did not interfere with the migration of LHRH neurons into the ophthalmic nerve bundle of the trigeminal nerve. The presence of structural supports seems to be primarily of importance in the migration of LHRH neurons along the olfactory and trigeminal nerve bundles. PSA may be less important for the migration of the LHRH neurons along peripheral neural elements.

Animals↗

Development of mossy fiber synapses in hippocampal slice culture.

The mossy fiber synaptogenesis has been studied in hippocampal slice cultures. In vivo mossy fiber terminals contact the thorny excrescences of CA3 pyramidal neurons over a restricted portion, i.e. the proximal part of the apical dendrite. In organotypic cultures mossy fibers expand their terminal field and invade the infrapyramidal area of the CA3 region and the supragranular layer of the dentate gyrus. Newly formed mossy fiber synapses in CA3 region were examined, through electron microscopy, in cultures taken at various time intervals. The main events of the formation of newly formed mossy fiber synapses can be summarized as follows. During the first week following explantation mossy fiber axons contact the dendritic shaft of the pyramidal dendrite and establish both symmetrical and asymmetrical contacts. Subsequent modifications occur in the postsynaptic portion facing the mossy fiber bouton: (i) a massive accumulation of polyribosomes and coated vesicles in the subsynaptic cytoplasm; (ii) undulations of the plasma membrane; (iii) disappearance of neurotubules at postsynaptic sites and appearance of a fine network of filamentous material. Later on in culture, complex giant spines invaginate within the synaptic bouton. In conclusion this study shows that CA3 pyramidal neurons following deafferentation retain the capacity to form thorny excrescences, when contacted by mossy fibers. Moreover these results suggest a crucial role for mossy fibers to induce the formation of thorny excrescences in an heterotopic localization, i.e. over the basilar dendrites of CA3 pyramidal neurons.

Animals↗