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Developmental ocular abnormalities in rats with X-ray--induced cataract mutation.

Prenatal and early postnatal devpelopment of eyes of Wistar rats with X-ray-induced cataract mutation was assessed histologically, to determine the events leading to cataract formation. A significant phenomenon appeared to be retinal folding, which occurred regularly at 14 to 15 days of gestation and might have pushed the lens against the cornea. A corneal reaction with the lens was indicated by corneal adhesions to the lens, seen frequently during and shortly after the period of retinal folding, and could have stimulated the epithelial hyperplasia that leads invariably to anterior polar cataract in these animals. Changes in the lens fiber cells, which could have been generated by the epithelial hyperplasia, included the sporadic detachment of newly developing fibers from the lens epithelium and the eventual swelling of the anterior ends of fibers still attached to the epithelium. The detached fibers became fusiform and developed postnatally into posterior, suture-associated vacuoles. Anterior uveitis was noted postnatally in some eyes and probably contributed to the subsequent development of the cataract. The results are discussed in the light of congenital anterior polar cataract formation reported in humans and other animals.

Animals↗

Postnatal expression of myosin heavy chains in muscle spindles of the rat.

The immunocytochemical expression of two myosin isoforms in intrafusal muscle fibers was examined in soleus muscles of neonatal (zero to six days postpartum) and adult rats. Monoclonal antibodies specific for myosin heavy chains of the slow-tonic anterior latissimus dorsi (ALD58) and fast-twitch pectoralis (MF30) muscles of the chicken were used. In adults ALD58 bound to the intracapsular regions of bag1 and bag2 fibers and MF30 bound to the intracapsular regions of bag2 and chain fibers. The extracapsular regions of intrafusal fibers and all extrafusal fibers did not react to ALD58 or MF30. Bag1 and bag2 fibers of neonatal rats expressed immature myosin patterns but chain fibers did not. The adult pattern of immunoreactivity of intrafusal fibers developed by the fourth postnatal day, when the patterns of motor but not sensory innervation in the spindle are still immature. Data suggest that the expression and maintenance of the specific anti-myosin immunoreactivity of intrafusal fibers during postnatal development of rat spindles is dependent upon sensory but not motor innervation. Moreover, afferents might regulate the gene expression responsible for synthesis of myosins isoforms specific to intrafusal fibers only in those myonuclei located within the capsule, but not in the myonuclei in extracapsular regions of intrafusal fibers.

Animals↗

Endothelial adherence under shear stress is dependent upon microfilament reorganization.

In response to externally applied shear stress, cultured endothelial monolayers develop prominent, axially-aligned, microfilamentous bundles, termed "stress fibers" (Dewey: Journal of Biomechanical Engineering 106:31-35, 1984; Franke et al.: Nature 81:570-580, 1984; Franke et al.: Klin. Wochenschr 64:989-992, 1986; Wechezak et al.: Laboratory Investigation 53:639-647, 1985). It is unclear, however, whether similar stress fibers develop in noncontiguous endothelial cells and whether these structures are necessary for adherence of individual cells under shear stress. It also is unknown what alterations occur in microtubules, intermediate filaments, and focal contacts as a consequence of shear stress. In this study, endothelial cells, free of intercellular contact, were exposed to 93 dynes/cm2 for 2 hr. With the aid of specific labeling probes and interference reflection microscopy, the distributional patterns of microfilaments, microtubules, intermediate filaments, and focal contacts were examined. Following shear stress, microfilament bundles and their associated focal contacts were concentrated in the proximal (relative to flow direction) cell regions. In contrast, microtubules were distributed uniformly within cell contours. Intermediate filaments displayed only an occasional tendency for accumulation at proximal edges. When cells were shear-tested in the presence of cytochalasin B to inhibit microfilament assembly, considerable cell loss occurred. Following inhibition of tubulin polymerization, no increase was observed in the percentage of cells lost due to shear over nontreated controls. Nocodazole-treated cells, however, were characterized by prominent stress fibers throughout the cell. These results indicate that stress fiber and focal contact reorganization represent major responses in isolated endothelial cells exposed to shear stress and that these cytoskeletal structures are necessary for adherence.

Actin Cytoskeleton↗

Development of intraosseous fibers of the mouse periodontium: a high-voltage electron microscopic study.

Development of intraosseous fibers was studied in mandibles of Swiss white mice, age 17 days to 45 days. Light microscopic (LM) and high-voltage electron microscopic (HVEM) techniques were used. In LM, Wilder's reticular stain revealed intraosseous fibers throughout the interdental septum by day 17. fibers were composed of unit collagen fibrils with abundant interfibrillar matrix. As development proceeded, fibers exhibited less interfibrillar matrix. Channels surrounding the intraosseous fibers became evident. Anchoring fibers attached the intraosseous fiber to bone. Fibroblasts appeared to be absent, suggesting that maintenance of the intraosseous fiber might be the function of the osteocyte. The presence of intraosseous fibers suggested a reevaluation of the fibrous attachment of teeth to bone. The concept of intraosseous fibers may simplify concepts of tooth movement and approximal drift.

Animals↗

Temporal and spatial patterns of twining force and lignification in stems of Ipomoea purpurea.

Using the TWIFOR, an electronic device for continuous, in vivo measurement of the forces exerted by twining vines, we examined the forces generated by vines growing on cylindrical poles of slender (6.35 mm) and thicker (19.05 mm) diameter. In stems of Ipomoea purpurea (L.) Roth. magnitudes of twining force (axial tensions) were, on average, less at a particular time and location on the more slender poles; while twining loads (normal force per unit length of vine) were much greater on the slender poles because of the greater curvature of the vines. Thus, the geometry of the helix formed by the vine on the pole affects the ability of the vine to maintain a frictional interaction with its support. In addition, the plant-to-plant variation in twining force was twice as great on the thicker support poles. Metaxylem and fibers developed closer to the plant apex in vines on the slender poles. On the thicker poles, a significant fraction of the maximum twining force developed during the establishment of the first gyre, before fibers were lignified, indicating that primary growth can be sufficient to establish high twining forces. On the slender poles, however, twining force increased with developmental stage until the gyre was at least 1.5 m from the apex. Thus, twining force can increase after cessation of primary growth. No simple relationship was found between the site of fiber differentiation and twining force.

Ipomoea↗

Role of calcium loading in early afterdepolarizations generated by Cs+ in canine and guinea pig Purkinje fibers.

INTRODUCTION: Our previous observations indicate that the Na+:Ca2+ exchange current (INa:Ca) plays an important role in early afterdepolarizations occurring at more negative Vm (L-EAD). The purpose of these studies was to examine the role of Ca(2+)-loading, which stimulates INa:Ca, in generation of L-EAD. METHODS AND RESULTS: Purkinje strands and preparations of ventricular myocardium from dogs and guinea pigs were superfused with oxygenated physiologic buffer solutions at 37 degrees C. To induce EADs, [K+]o was reduced to 2.0 to 3.0 mM and [Cs+]o (3.6 to 4.0 mM) was added at slow rates of < or = 0.3 Hz. Isometric contraction in canine Purkinje strands and guinea pig papillary muscles doubled in 1-hour exposure to Cs+ and low [K+]o at slow rates and the uptake of 45Ca2+ was approximately doubled after 30 minutes. Forty-three percent of Purkinje fibers developed L-EAD after a latent period of 17 to 123 minutes of exposure. Ouabain (0.2 microM) suppressed L-EAD within 10 minutes reversibly. Ca(2+)-loading (low [Na+]o or high [Ca2+]o) for 5 to 10 minutes before exposure to Cs+, low [K+]o, and slow rates resulted in rapid development of L-EAD in all preparations during subsequent exposure. In Ca(2+)-loaded preparations, delayed afterdepolarizations (DADs) as well as L-EADs developed. CONCLUSIONS: Reduction of K+ currents with Cs+, low [K+]o, and slow rates induced L-EAD in a fraction of Purkinje fibers after a latent period during which Ca(2+)-loading of the sarcoplasmic reticulum occurred, while fibers preloaded with Ca2+ developed L-EAD rapidly and uniformly. These findings indicate that Ca(2+)-loading is a critical condition for the development of L-EAD. Early suppression of L-EAD by ouabian suggests a dependence of L-EAD on low [Na+]i. These findings implicate INa:Ca in the generation of L-EAD.

Animals↗

The role of dietary fiber in the development and treatment of childhood obesity.

Childhood obesity is viewed as a public health problem in the United States because of its assumed high prevalence and increasing secular trend. The best estimate of the genetic contribution to obesity ranges from 5% to 25%. Environmental factors play a major role in obesity development. Low income and a low level of education have been associated with obesity, particularly among white women. Caloric intake as a risk factor for obesity has not been clearly established. This lack of a clear-cut association may be attributable to the problem of accurately measuring caloric intake. Several studies have linked increased total fat intake, rather than caloric intake, with obesity. Some studies have linked television viewing to obesity in children. Obesity is rare among the populations of developing countries, where dietary fiber intake is high. Explanations for the role of dietary fiber in obesity include a reduced caloric density of the foods, a slower rate of food ingestion, and possible effects on satiety. Most studies on the role of fiber in the treatment of obesity have been somewhat limited by lack of comparison groups, inadequate sample sizes, and short durations of the observations. However, although limited, the available evidence suggests that fiber potentially could play a useful role in weight reduction. For children, fiber administration should be considered as an adjuvant therapy rather than a primary modality, because fiber might aid in promoting satiety during meals and curbing hunger between meals.(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗

Lanceolate hair-J (lahJ): a mouse model for human hair disorders.

Lanceolate hair-J (lahJ) arose spontaneously in 1994 on the DBA/1LacJ inbred background at The Jackson Laboratory. Mutant mice were runted, alopecic, and lacked vibrissae. As they aged, their skin wrinkled. Affected mice developed a noninflammatory, proliferative skin disease with follicular dystrophy. Hair fibers developed a number of abnormalities including periodic nodules along the shaft (trichorrhexis nodosa), compaction resembling trichorrhexis invaginata, spiral fractures, broken tips, and lance-shaped tips. This mutation exhibits some characteristics that resemble an autosomal recessive ichthyosiform disease that occurs in humans characterized in part by peculiar, invaginating, multinodal, hair shaft abnormalities known as Netherton's syndrome. Periodic nodules also resemble the human genetic based disease monilethrix. This autosomal recessive mouse mutation, allelic with lanceolate hair (lah), based on breeding studies, is located on mouse Chromosome 18, within a cluster of genes coding for adhesion molecules. Homozygotes for either of these allelic mouse mutations have elevated serum IgE levels, a feature also common with human Netherton's syndrome.

Alleles↗

Expression of GAP-43 in growing efferent fibers during cochlear development.

The development of olivocochlear efferent fibers has been studied by means of the immunocytochemical detection of the growth-associated protein GAP-43. This study has been carried out in pre- and postnatal, developing, pigmented rats. Results indicate that olivocochlear efferents reach the developing auditory epithelium from embryonic day 18 on. GAP-43-like immunoreactivity persists in the organ of Corti until the beginning of the second postnatal month. The relationships between the expression of GAP-43 in olivocochlear efferent fibers and other plasticity-related phenomena are discussed.

Animals↗

Development of fungiform papillae, taste buds, and their innervation in the hamster.

Fungiform taste buds in mature hamsters are less subject to neurotrophic influences than those of other species. This study evaluates taste-bud neurotrophism during development in hamsters by examining the relation between growing nerves and differentiating fungiform papillae. Chorda tympani (CT) or lingual (trigeminal) nerve (LN) fibers were labelled with Lucifer Yellow as they grew into (CT fibers) or around (LN fibers) developing taste buds. Developing fungiform papillae and taste pores were counted with the aid of a topical tongue stain. The tongue forms on embryonic days (E) 10.5-11 and contains deeply placed CT and LN fibers but no papillae. By E12, the tongue epithelium develops scattered elevations. These "eminences" selectively become innervated by LN fibers that grow to the epithelium earlier and in larger numbers than CT fibers. Definitive fungiform papillae form rapidly during E13-14 and become heavily innervated by LN fibers. Intraepithelial CT fibers, rare at E13, invariably innervate fungiform papillae containing nascent taste buds at E14. During E14-15 (birth = E15-16), most papillae contain taste buds with pores, extensive perigemmal LN innervation, and extensive intragemmal CT innervation. At birth, numbers of fungiform papillae and taste pores are adultlike. The results show that fungiform eminences begin forming in the absence of innervation. The subsequent differentiation of definitive fungiform papillae and their innervation by LN fibers occur synchronously, prior to the differentiation of taste buds and their CT innervation. The hamster is precocious (e.g., compared to rat) in terms of LN development and the structural maturity of the anterior tongue at birth.

Animals↗

Effect of early gestation feeding, birth weight, and gender of progeny on muscle fiber characteristics of pigs at slaughter.

Maternal nutrition and progeny birth weight affect muscle fiber development in the pig, thereby influencing early postnatal growth rate. The objective of the study was to determine the extent to which growth, morphometric characteristics, and area and distribution of slow-oxidative (SO), fast oxidative-glycolytic (FOG), and fast glycolytic (FG) fibers of three muscles (LM = longissimus muscle; RF = rectus femoris; ST = semitendinosus) of slaughter pigs were affected by DE intake level during the first 50 d of gestation. Multiparous Swiss Large White sows were assigned randomly to one of three energy intake treatments: 1) fed 2.8 kg/d of a standard diet (STD; n = 6) containing 10.7 MJ DE/kg; 2) fed 2.8 kg/d of a low-energy diet (LE; n = 5) containing 6.6 MJ DE/kg; or 3) fed 4.0 kg/d of a standard diet (HE; n = 5) containing 10.7 MJ DE/kg (as-fed basis). Sows were subjected to energy intake treatments for the first 50 d of gestation; however, from d 51 to parturition, sows received 2.8 kg/d of the standard diet, and the amount of feed offered each sow during lactation was adjusted according to the litter size. Sows farrowed normally and pig birth weights were recorded. Based on birth weight, the two lightest (1.27 kg; Lt) and two heaviest (1.76 kg; Hvy) barrows and gilts from the 16 litters (n = 64) were selected at weaning and were offered a fixed amount of feed (170 g x BW(0.569)/d) from 25 to 105 kg BW. Regardless of the birth weight, progeny from HE sows grew slower (P < 0.05) during lactation and the growing-finishing period, had a lower (P < 0.05) gain-to-feed ratios, and had higher (P < 0.05) percentages of adipose tissue than pigs born from LE sows. The ST was shorter (P = 0.03) in Lt than in Hvy pigs, and the ST of gilts was heavier (P = 0.01) and had a larger (P = 0.01) girth than the ST of barrows. Overall mean fiber area tended to be larger (P < or = 0.11) in the LM and light portion of the ST of Lt than in Hvy pigs, and was larger (P = 0.03) in the ST of gilts than barrows. The ST of progeny from LE sows had fewer (P < 0.10) FG fibers, which was compensated by either more (P < 0.05) FOG in the light portion of the ST, or more (P < 0.10) SO fibers in the dark portion, and these differences were more pronounced in Lt pigs than in Hvy pigs. Overall, maternal feeding regimen affected muscle fiber type distribution, whereas birth weight and gender affected muscle fiber area.

Animal Feed↗

Syncoilin upregulation in muscle of patients with neuromuscular disease.

Syncoilin may have a role in linking the desmin-associated intermediate filament network of the muscle fiber with the dystrophin-associated protein complex (DAPC). We have evaluated syncoilin in a range of neuromuscular disorders including Duchenne and Becker muscular dystrophy, central core disease, congenital muscular dystrophies, and neurogenic disorders. Our results show that syncoilin immunolabeling is not only altered in muscle fibers with alterations in the DAPC but also in response to a variety of genetic defects, including those associated with proteins of the extracellular matrix and the intracellular Ca2+-release channel (ryanodine receptor). The pattern of syncoilin immunolabeling in these diseases appeared to reflect a rearrangement of the intermediate filament-associated cytoskeleton that characterizes both muscle fiber development and conditions in which the cytoskeletal organization of the muscle fiber is significantly affected. These observations raise the possibility that mutations in the gene encoding for syncoilin may underlie some forms of muscle disease.

Animals↗

The MLC1v gene provides a transgenic marker of myocardium formation within developing chambers of the Xenopus heart.

Many details of cardiac chamber morphogenesis could be revealed if muscle fiber development could be visualized directly within the hearts of living vertebrate embryos. To achieve this end, we have used the active promoter of the MLC1v gene to drive expression of green fluorescent protein (GFP) in the developing tadpole heart. By using a line of Xenopus laevis frogs transgenic for the MLC1v-EGFP reporter, we have observed regionalized patterns of muscle formation within the ventricular chamber and maturation of the atrial chambers, from the onset of chamber formation through to the adult frog. In f1 generation MLC1v-EGFP animals, promoter activity is first detected within the looping heart tube and delineates the forming ventricular chamber and proximal outflow tract throughout their development. The 8-kb MLC1v promoter faithfully reproduces the embryonic expression of the endogenous MLC1v mRNA. At later larval stages, weak patches of EGFP fluorescence are found on the atrial side of the atrioventricular boundary. Subsequently, an extensive lattice of MLC1v-expressing fibers extend across the mature atrial chambers of adult frog hearts and the transgene reveals the differing arrangement of muscle fibers in chamber versus outflow myocardium. The complete activity of the promoter resides within the proximal 4.5 kb of the MLC1v DNA fragment, whereas key elements regulating chamber-specific expression are present in the proximal-most 1.5 kb. Finally, we demonstrate how cardiac and craniofacial muscle expression of the MLC1v promoter can be used to diagnose mutant phenotypes in living embryos, using the injection of RNA encoding a Tbx1-engrailed repressor-fusion protein as an example.

Animals↗

Development of chicken intrafusal muscle fibers.

The first sign of developing intrafusal fibers in chicken leg muscles appeared on embryonic day (E) 13 when sensory axons contacted undifferentiated myotubes. In sections incubated with monoclonal antibodies against myosin heavy chains (MHC) diverse immunostaining was observed within the developing intrafusal fiber bundle. Large primary intrafusal myotubes immunostained moderately to strongly for embryonic and neonatal MHC, but they were unreactive or reacted only weakly with antibodies against slow MHC. Smaller, secondary intrafusal myotubes reacted only weakly to moderately for embryonic and neonatal MHC, but 1-2 days after their formation they reacted strongly for slow and slow-tonic MHC. In contrast to mammals, slow-tonic MHC was also observed in extrafusal fibers. Intrafusal fibers derived from primary myotubes acquired fast MHC and retained at least a moderate level of embryonic MHC. On the other hand, intrafusal fibers developing from secondary myotubes lost the embryonic and neonatal isoforms prior to hatching and became slow. Based on relative amounts of embryonic, neonatal and slow MHC future fast and slow intrafusal fibers could be first identified at E14. At the polar regions of intrafusal fibers positions of nerve endings and acetylcholinesterase activity were seen to match as early as E16. Approximately equal numbers of slow and fast intrafusal fibers formed prenatally; however, in postnatal muscle spindles fast fibers were usually in the majority, suggesting that some fibers transformed from slow to fast.

Animals↗

Effect of fiber optic probe geometry on depth-resolved fluorescence measurements from epithelial tissues: a Monte Carlo simulation.

Developing fiber optic probe geometries to selectively measure fluorescence spectra from different sublayers within human epithelial tissues will potentially improve the endogenous fluorescence contrast between neoplastic and nonneoplastic tissues. In this study, two basic fiber optic probe geometries, which are called the variable aperture (VA) and multidistance (MD) approaches, are compared for depth-resolved fluorescence measurements from human cervical epithelial tissues. The VA probe has completely overlapping illumination and collection areas with variable diameters, while the MD probe employs separate illumination and collection fibers with a fixed separation between them. Monte Carlo simulation results show that the total fluorescence detected is significantly higher for the VA probe geometry, while the probing depth is significantly greater for the MD probe geometry. An important observation is that the VA probe is more sensitive to the epithelial layer, while the MD probe is more sensitive to the stromal layer. The effect of other factors, including numerical aperture (NA) and tissue optical properties on the fluorescence measurements with VA and MD probe geometries, are also evaluated. The total fluorescence detected with both probe geometries significantly increases when the fiber NA is changed from 0.22 to 0.37. The sensitivity to different sublayers is found to be strongly dependent on the tissue optical properties. The simulation results are used to design a simple fiber optic probe that combines both the VA and MD geometries to enable fluorescence measurements from the different sublayers within human epithelial tissues.

Absorption↗

Ca-, Sr-tension relationships and contraction velocities of human muscle fibers.

Muscle fibers from the lateral gastrocnemius or intercostal muscles of 7 normal adult males were chemically skinned (sarcolemma disrupted) and isolated fibers were divided into two parts for histochemical determination of fiber type and physiologic studies. The Ca- and Sr-induced tension relationships and maximum contraction velocities were measured. Slow twitch fibers developed tension at lower concentrations of Ca or Sr than fast twitch fibers. The difference between fast and slow twitch fibers was greatest when Sr was the activating cation. Fast and slow twitch fibers was greatest when Sr was the activating cation. Fast and slow twitch fibers generated similar maximum tensions. The contraction velocities of fast twitch fibers were more than two-fold greater than slow twitch fibers. Fast-oxidative-glycolytic (FOG, type IIA) and fast-glycolytic (FG, type IIB) fibers had similar Ca- and Sr-tension relationships and contraction velocities.

Adult↗

The Arabidopsis RHD3 gene is required for cell wall biosynthesis and actin organization.

The Arabidopsis thaliana (L.) Heynh. ROOT HAIR DEFECTIVE3 (RHD3) gene has previously been shown to be essential for normal cell expansion [H. Wang et al. (1997) Genes Dev 11:799-811]. In this report, we demonstrated that mutation of the RHD3 gene in the Arabidopsis fragile fiber 4 (fra4) mutant caused a dramatic reduction in the wall thickness of fibers, vessels, and pith cells in the inflorescence stems and, concomitantly, a decrease in the mechanical strength of stems. The reduced wall thickness in the fra4 mutant was accompanied by an alteration in cell wall composition. Consistent with the defective fiber and vessel wall phenotypes, the RHD3 gene exhibited a strong expression in developing fiber and xylem cells. We showed that the Arabidopsis genome contains two additional RHD3-like genes, one of which was expressed specifically in flowers. In addition, we found that mutation of the RHD3 gene caused an alteration in the organization of the actin cytoskeleton but no effects on cortical microtubules. Our findings suggest an essential role of RHD3 in cell wall biosynthesis and actin organization, both of which are known to be important for cell expansion.

Actins↗

Fibulin-5 mutations: mechanisms of impaired elastic fiber formation in recessive cutis laxa.

To elucidate the molecular mechanisms of impaired elastic fiber formation in recessive cutis laxa, we have investigated two disease-causing missense substitutions in fibulin-5, C217R and S227P. Pulse-chase immunoprecipitation experiments indicated that S227P mutant fibulin-5 was synthesized and secreted by skin fibroblasts at a reduced rate when compared with the wild-type protein. Both mutants failed to be incorporated into elastic fibers by transfected rat lung fibroblasts. Purified recombinant fibulin-5 with either mutation showed reduced affinity for tropoelastin in solid-phase binding assays. Furthermore, S227P mutant fibulin-5 also showed impaired association with fibrillin-1 microfibrils. The same mutation triggered an endoplasmic reticulum (ER) stress response, as indicated by the strong co-localization of this mutant protein with folding chaperones in the ER, including calreticulin, immunoglobulin-binding protein and protein disulfide isomerase, and by increased rates of apoptosis in patient fibroblasts. Histological analysis of skin sections from a cutis laxa patient with a homozygous S227P mutation showed a lack of fibulin-5 in the extracellular matrix and a concomitant disorganization of dermal elastic fibers. By electron microscopy, elastic fibers in the skin of this patient showed a failure of elastin globules to fuse into a continuous elastic fiber core. We conclude that recessive cutis laxa mutations in fibulin-5 result in misfolding, decreased secretion and a reduced interaction with elastin and fibrillin-1 leading to impaired elastic fiber development. These findings support the hypothesis that fibulin-5 is necessary for elastic fiber formation by facilitating the deposition of elastin onto a microfibrillar scaffold via direct molecular interactions.

Amino Acid Substitution↗