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PubMed2006

Molecular characterization and expression analysis of nine cotton GhEF1A genes encoding translation elongation factor 1A.

The translation elongation factor 1A, eEF1A, plays an important role in protein synthesis, catalyzing the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. To investigate the role of eEF1A for protein synthesis in cotton fiber development, nine different cDNA clones encoding eukaryotic translation elongation factor 1A were isolated from cotton (Gossypium hirsutum) fiber cDNA libraries. The isolated genes (cDNAs) were designated cotton elongation factor 1A gene GhEF1A1, GhEF1A2, GhEF1A3, GhEF1A4, GhEF1A5, GhEF1A6, GhEF1A7, GhEF1A8, GhEF1A9, respectively. They share high sequence homology at nucleotide level (71-99% identity) in the coding region and at amino acid level (96-99% identity) among each other. Phylogenetic analysis demonstrated that the nine GhEF1A genes can be divided into 5-6 subfamilies, indicating the divergence occurred in structures of the genes as well as the deduced proteins during evolution. Real-time quantitative RT-PCR analysis revealed that GhEF1A genes are differentially expressed in different tissues/organs. Of the nine GhEF1A genes, five are expressed at relatively high levels in young fibers. Further analysis indicated that expressions of the GhEF1As in fiber are highly developmental-regulated, suggesting that protein biosynthesis is very active at the early fiber elongation.

Amino Acid Sequence↗

PubMed1961

The ionic mechanisms of hyperpolarizing responses in lobster muscle fibers.

Lobster muscle fibers develop hyperpolarizing responses when subjected to sufficiently strong hyperpolarizing currents. In contrast to axons of frog, toad, and squid, the muscle fibers produce their responses without the need for prior depolarization in high external K(+). Responses begin at a threshold polarization (50 to 70 mv), the potential reaching 150 to 200 mv hyperpolarization while the current remains constant. The increased polarization develops at first slowly, then becomes rapid. It usually subsides from its peak spontaneously, falling temporarily to a potential less hyperpolarized than at threshold for the response. As long as current is applied there can be oscillatory behavior with sequential rise and subsidence of the polarization, repeating a number of times. Withdrawal of current leads to rapid return of the potential to the resting level and a small, brief depolarization. Associated with the latter, but of longer duration, is an increased conductance whose magnitude and duration increase with the antecedent current. Hyperpolarizing responses of lobster muscle fibers are due to increased membrane resistance caused by hyperpolarizing K inactivation. The oscillatory characteristic of the response is due to a delayed superimposed and prolonged increase in membrane permeability, probably for Na(+) and for either K(+) or Cl(-). The hyperpolarizing responses of other tissues also appear to result from hyperpolarizing K inactivation, on which is superimposed an increased conductance for some other ion or ions.

Animals↗

PubMed1982

Electrophysiological correlates of rapid escape reflexes in intact earthworms, Eisenia foetida. I. Functional development of giant nerve fibers during embryonic and postembryonic periods.

Grids of recording electrodes etched onto printed circuit boards were used for noninvasive recording of medial (MGF) and lateral (LGF) giant nerve fiber spikes in developing earthworms, Eisenia foetida. Stereotyped patterns of through-conducted giant fiber spikes, evoked by light tactile stimulation, were first detectable in the normal crawling embryonic stage and continued to be detectable throughout postembryonic development. Giant fiber spiking activity in normal crawling embryos was accompanied by stereotyped muscle activity and rapid escape withdrawal, suggesting that giant fiber reflex pathways are functionally intact before the worm hatches. For both the MGF and LFG, several age-dependent changes were noted, including the following: increases in spike conduction velocity, increases in giant fiber diameter, and decreases in spike duration. The MGF conduction velocity in normal crawling embryos was 1.1-1.6 m s-1 (6-7 micrometers diameter) and increased to 7.0-8.5 m s-1 (20-25 micrograms diameter) by 60 days after hatching. The LGF conduction velocity in normal crawling embryos was 0.7-1.1 m s-1 (2.5-4.0 micrometers diameter) and increased to 4.0-5.5 m s-1 (8-14 micrometers diameter) by 60 days after hatching. During postembryonic development MGF and LGF conduction velocities were linearly related to fiber diameter.

Aging↗

PubMed2002

Development of neuromuscular junctions in the mouse esophagus: focus on establishment and reduction of enteric co-innervation.

The development of vagal and enteric innervation of esophageal motor endplates was examined in perinatal and adult BALB/c and NMRI mice using immunocytochemistry and confocal laser scanning microscopy. Nicotinic acetylcholine receptors were demonstrated with fluorochrome-tagged alpha-bungarotoxin, vagal motor terminals with antisera against vesicular acetylcholine transporter and calcitonin gene-related peptide, and enteric nerve terminals with antisera against neuronal nitric oxide synthase, vasoactive intestinal peptide and galanin. Results demonstrated that enteric and vagal innervations of striated esophageal muscle fibers develop in close spatiotemporal relationship, but with different courses. Connections between VAChT-positive vagal nerve terminals and growing acetylcholine receptor clusters were established from E17 to reach 100% motor endplate innervation at P14 and were maintained throughout adult life. CGRP immunoreactivity developed with a delay of several days after the appearance of VAChT in vagal terminals. From P14 to adulthood CGRP was colocalized with VAChT in almost all motor endplates. In contrast, enteric co-innervation rates increased from E17 to a maximum of 70-80% at P4, while their incidence at motor endplates progressively declined over the following 5 months to lower levels maintained throughout adulthood. Whereas adult enteric co-innervation rates in BALB/c and NMRI mice differed significantly (approximately 30% versus approximately 10%, respectively), their increase and reduction, respectively, during development showed an identical time course. These results suggest a well-ordered sequence of attraction of enteric nerve fibers to, and removal from motor endplates in the developing mouse esophagus. Thus, enteric co-innervation may subserve a functional role in the development and control of perinatal striated esophageal muscle rather than representing an unspecific "hangover" from the smooth muscle past of this organ.

Animals↗

PubMed1998

Differences in myelination between spinal cord and corticular tissue transplanted intraocularly in rats.

This paper compares the myelination in rat cortex and spinal cord transplanted on the embryonic day (E) 12, 14, 16, 18, 20 and right after birth (P0) into the anterior eye chamber of adult rats. Myelinated fibers were not observed in either cortical or spinal cord transplants of E12. When transplanted on E14, abundant myelinated fibers developed in the spinal cord and gathered at the periphery of the transplants as a "white matter". In the grafted cortex myelinated fibers were found when transplantation occurred on E16 or later. The myelinated fibers, however, remained scarce or formed only narrow bundles. The number and distribution of myelinated fibers did not depend on the donor's age between E16 and P0: even in the latter case transplanted cortex were found without myelination. The differences could be attributed to the different development of the cortical and spinal cord oligodendrocytes and to the different intrinsic organization of the grafted samples.

Animals↗

PubMed1990

Characteristics of A- and C-fibers ending in a sensory nerve neuroma in the rat.

1. We have studied, in vivo, the degree of spontaneous activity, responsiveness to mechanical and chemical stimuli, and the conduction velocities in C- and A-fibers ending in the neuromas formed 8-66 days after ligation and transection of a cutaneous sensory nerve in the rat. 2. Some of these C- and A-fibers developed ongoing activity. The percentage varied considerably between neuromas in different animals, from 0 to 23% (mean, 4.2%), with no major variation in the incidence as a function of neuroma age. 3. The endings of the fibers in the neuroma could be excited by both mechanical and chemical stimuli. From 0 to 26% (mean, 13%) of these fibers had mechanosensitive endings, some of which were located in the muscle/facia tissue outside the neuroma itself. Some fibers were excited by direct application of chemicals to their endings in the neuroma; 3.0% of A- and C-fibers responded to bradykinin, 2.0% to histamine, and 2.8% to adrenaline. There was no systemic variation in the percentages of mechano- or chemosensitive fibers with neuroma age. 4. The C-fiber action potentials showed a continuing decrease in conduction velocities over the 9 wk after nerve transection. More than 4 wk after transection, the conduction velocity of neuroma fibers was 88% that of C-fibers of normal saphenous nerve. 5. We conclude that fibers in a cutaneous nerve neuroma have some sensory capabilities similar to those in normal nerves terminating in the skin. This could be because they are retained after the nerve is transected or because they are initially lost but then regenerate. However, the numbers are restricted, probably because the fibers remain isolated from factors produced by their target skin tissue that are necessary for development and maintenance of sensory functions.

Action Potentials↗

PubMed2004

Hedgehog signaling is required for commitment but not initial induction of slow muscle precursors.

In zebrafish, skeletal muscle precursors can adopt at least three distinct fates: fast, non-pioneer slow, or pioneer slow muscle fibers. Slow muscle fibers develop from adaxial cells and depend on Hedgehog signaling. We analyzed when precursors become committed to their fates and the step(s) along their differentiation pathway affected by Hedgehog. Unexpectedly, we find that embryos deficient in Hedgehog signaling still contain postmitotic adaxial cells that differentiate into fast muscle fibers instead of slow. We show that by the onset of gastrulation, slow and fast muscle precursors are already spatially segregated but uncommitted to their fates until much later, in the segmental plate when slow precursors become independent of Hedgehog. In contrast, pioneer and non-pioneer slow muscle precursors share a common lineage from the onset of gastrulation. Our results demonstrate that slow muscle precursors form independently of Hedgehog signaling and further provide direct evidence for a multipotent muscle precursor population whose commitment to the slow fate depends on Hedgehog at a late stage of development when postmitotic adaxial cells differentiate into slow muscle fibers.

Animals↗

PubMed1994

Stimulation of in vitro myelin synthesis by microglia.

Central nervous system myelin is elaborated by oligodendrocytes, which have been studied extensively in cell culture. Dissociated brain cultures allow in vitro analysis of events in myelinogenesis, including cell-cell interactions. Microglia, the primary phagocytic cell of the central nervous system, appear in developing fiber tracts prior to the onset of myelination in vivo. To gain insight into potential oligodendrocyte-microglial interactions during development, these cells were co-cultured and various parameters of myelin synthesis were measured. In co-culture, microglia stimulated the synthesis of sulfatide, a myelin-specific galactolipid, in oligodendrocytes, as well as the expression of the myelin-specific proteins myelin basic protein and proteolipid protein. Activity of the oligodendrocyte cytoplasm-specific enzyme 2',3'-cyclic nucleotide 3'-phosphohydrolase was not elevated, suggesting that the effects of microglia were not due to stimulation of oligodendrocyte proliferation. This was confirmed by the inability of microglia to induce significant DNA synthesis. Conditioned medium from cultured microglia provided a similar stimulatory activity, suggesting that the increase in myelin synthesis does not require contact between oligodendrocytes and microglia. These findings suggest a stimulatory role for microglia during myelinogenesis.

2',3'-Cyclic Nucleotide 3'-Phosphodiesterase↗

PubMed1982

The in-vitro caffeine contracture test: influence of the muscle histochemical profile on test results.

In vitro caffeine contracture tests were carried out on whole rat muscle composed primarily of either histochemical type I or type II fibers. Muscles composed primarily of type I fibers developed contractures at lower concentrations of caffeine and had lower caffeine specific concentrations than muscles composed primarily of type II fibers. These findings indicate that the histochemical profile of a muscle can influence the results of the in-vitro caffeine contracture test.

Animals↗

PubMed2003

[Cloning and characterization of the PTS2 receptor gene (GhPex7) from cotton (Gossypium hirsutum L.)].

Peroxisomal targeting signals (PTS), including PTS1 and PTS2, were proposed to play an important role in introducing proteins into the peroxisomal matrix. Pex7, the PTS2 receptor, is crucial to the import of PTS2 proteins. Based on the sequence of a fragment (F010) recovered from cDNA-AFLP, the full-length cDNA sequence was obtained by RACE and the contig in cotton ESTs, and the coding sequence was further cloned. The GhPex7 cDNA, 1314 bp in length, contained 5 non-coding (77 bp) upstream, 3 complete downstream with polyA signal tail and ORF of 954 bp, which coded for a deduced protein of 317 amino acids. The deduced protein had a predicted MW 35.57 kDa and a pI of 5.603. Homology analysis demonstrated that GhPex7 protein contained three highly conserved domains and three G-bata domains of WD-40 proteins family. The sequence of nucleotides and amino acids shared 83% and 76% identity with known Arabidopsis AtPex7, respectively, and its amino acid sequence shared from 28% to 42% identity with those of Drosophila melanogaster, Saccach, cerevisiae, Mus musculus and Homo sapiens. Southern blotting suggested that at least two copies of GhPex7 gene existed in Gossypium hirstum genome. By Northern blotting and RT-PCR analysis, expression of the GhPex7 was detected in roots, stems, leaves, buds, ovules and fibers, however it was stronger in leaves and stems than in other tissues. At the stage of cotton's ovules and fibers development, RT-PCR analysis also indicated that expression activity of GhPex7 in ovule at 0DPA mutant plant (no-fiber) was stronger than in wild type plant, its expression in wild type fiber at 23DPA was stronger than at 12, 16 DPA too.

Amino Acid Sequence↗

PubMed2004

Production and characterization of chitosan fibers and 3-D fiber mesh scaffolds for tissue engineering applications.

This study reports on the production of chitosan fibers and 3-D fiber meshes for the use as tissue engineering scaffolds. Both structures were produced by means of a wet spinning technique. Maximum strain at break and tensile strength of the developed fibers were found to be 8.5% and 204.9 MPa, respectively. After 14 d of immersion in simulated body fluid (SBF), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and inductively coupled plasma emission (ICP) spectroscopy analyses showed that a bioactive Ca-P layer was formed on the surface of the fibers, meaning that they exhibit a bioactive behavior. The samples showed around 120% max. swelling in physiological conditions. The pore sizes of 3-D chitosan fiber mesh scaffolds were observed to be in the range of 100-500 microm by SEM. The equilibrium-swelling ratio of the developed scaffolds was found to be around 170% (w/w) in NaCl solution at 37 degrees C. Besides that, the limit swelling strain was less than 30%, as obtained by mechanical spectroscopy measurements in the same conditions. The viscoelastic properties of the scaffolds were also evaluated by both creep and dynamic mechanical tests. By means of using short-term MEM extraction test, both types of structures (fibers and scaffolds) were found to be non-cytotoxic to fibroblasts. Furthermore, osteoblasts directly cultured over chitosan fiber mesh scaffolds presented good morphology and no inhibition of cell proliferation could be observed.Osteoblast-like cells proliferating over chitosan based fibers after 7 d of culture.

Animals↗

PubMed2007

Functional analysis of a cotton glucuronosyltransferase promoter in transgenic tobaccos.

The 5' fragment (1 647 bp) of the cotton glucuronosyltransferase gene (GhGlcAT1) was transcriptionally fused to the beta-glucuronidase (GUS) gene, and functionally analyzed for important regulatory regions controlling gene expression in transgenic tobacco plants. GUS activity analysis revealed that the full-length promoter drives efficient expression of the GUS gene in the root cap, seed coat, pollen grains and trichomes. Exposure of the transgenic tobacco to various abiotic stresses showed that the promoter was mainly responsive to the sugars (glucose and sucrose) as well as gibberellic acid. Progressive upstream deletion analyses of the promoter showed that the region from -281 to +30 bp is sufficient to drive strong GUS expression in the trichomes of shoot, suggesting that the 311 bp region contains all cis-elements needed for trichome-specific expression. Furthermore, deletion analysis also revealed that the essential cis-element(s) for sucrose induction might be located between -635 and -281 bp. In addition, sequence analysis of the regulatory region indicated several conserved motifs among which some were shared with previously reported seed-specific elements and sugar-responsive elements, while others were related with trichome expression. These findings indicate that a 1 647-bp fragment of the cotton GhGlcAT1 promoter contains specific transcription regulatory elements, and provide clues about the roles of GhGlcAT1 in cotton fiber development. Further analyses of these elements will help to elucidate the molecular mechanisms regulating the expression of the GhGlcAT1 gene during fiber elongation.

Base Sequence↗

PubMed1989

[The development of spinal endoscope using a flexible optic fiber].

The development of spinal endoscope using a thin (0.75 mm in diameter) flexible fiber catheter AS-001, Fukuda-densi Co Ltd., Japan) is described in this study. This fiber catheter contains 3,000 optic fibers as imaging and illuminating fibers within its diameter. The effective length and the visual angle is 1.10 m and 53 degree, respectively. Video processor system was connected to this fiber catheter. With a patient in the lateral decubitus position under local anesthesia, this fiber catheter was introduced into the lumbar subarachnoid space in a similar manner as lumbar spinal drainage. Eight patients with myelopathy were evaluated by this spinal endoscopic study. With other radiological diagnostic modalities such as myelography, selective spinal angiography using intraarterial digital subtraction angiography (DSA), or magnetic resonance imaging (MRI), it was difficult to make definitive diagnosis in these 8 patients. In 4 patients among them, vascular tangle, or tortuous course of the dilated vessels were visualized on the dorsal surface of the spinal cord by the spinal endoscopic study, which strongly suggested the spinal arteriovenous malformation. In 3 of them, operative findings verified these preoperative endoscopic diagnoses to be correct. In the remaining case, surgical intervention was not attempted because of the personal affair of the patient. In every case, several structures around the thoracolumbar cord such as spinal dura mater, arachnoid membrane, or conus medullaris were observed under the pulsation of the cerebrospinal fluid. No complication was associated with this spinal endoscopic study. About 10 minutes were required for the whole procedure.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1982

[Noradrenergic innervation of rat neocortex during development: immunocytochemical evidence using anti-dopamine-beta-hydroxylase antibodies].

Dopamine-beta-hydroxylase (DBH), the synthetizing enzyme for norepinephrine, has been visualized by immunofluorescence technique ontogeny in the Rat cerebral cortex. DBH like immunoreactive fibers develop as two bundles, one in the marginal zone observed at embryonic day 19 and another in the deep cortex visible at birth. These plexuses rapidly invade the whole neocortex following a rostro-caudal gradient. Five days after birth, all cortical layers show DBH containing fibers but the adult pattern is only reached two weeks after birth.

Aging↗

PubMed1994

Neogenesis of skeletal muscle in the postinflammatory rat peritoneum.

During the healing phase of a chemical peritonitis, one or several skeletal muscle fibers develop, de novo, in the peritoneum of the adult of weanling rat diaphragm. One week after the initial injury the new muscle fibers are narrow and have central nuclei and cross-striations. The fibers increase progressively in caliber and the nuclei take up a subsarcolemmal location. The newly formed muscle fibers are separated from the intrinsic diaphragmatic muscle by an elastic membrane and by a hand of hyaline connective tissue. They are separated from the peritoneal surface by a zone of granulation tissue. Most new fibers are oriented at right angles to the intrinsic diaphragmatic muscle fibers. Their location and orientation suggest an origin from mesothelium or from fibroblasts in the granulation tissue rather than from the intrinsic diaphragmatic muscle. A similar phenomenon can be induced in the pleura on the other side of the diaphragm. In contrast, damage to the diaphragmatic muscle by injection of aluminum lactate does not engender myogenesis in the location described despite active regeneration in the intrinsic muscle.

Animals↗

PubMed2003

Evidence for an extensive collagen type III/VI proximal domain in the rat femur. I. Diminution with ovariectomy.

Collagenous proteins other than Type I have received little attention in hypogonadal bone loss. Using femora from 25 young (2.5 months) and older (11 months) control and ovariectomized adult rats killed 1-4 months postoperation, cancellous atrophy was histologically confirmed, and the immunolocalization of collagen Type III was examined. This occurred as numerous immunofluorescent Sharpey-like fibers, 5-25 microm thick, regularly associated with collagen Type VI, which ramified the femoral cortex. Sequential transverse cryosections enabled the mapping of the fibers in three-dimensions, demonstrating that they constituted an extensive subperiosteal domain which may be a lasting legacy of early skeletal development. Fiber density was greatest in the trochanters and femoral neck. The domain tapered distally and was apparently anchored into the mid-shaft by intracortical cartilaginous islands, staining for collagen Type VI (as well as Type II and fibronectin). Ovariectomy caused disconnection of the fibers and reduced the proximal domain of both young and older animals, previously positive areas of the cortex becoming negative. It is concluded that collagen Type III/VI occupies a substantial, discrete domain in the rat proximal femur as a complex extension of the periosteum. Diminution of this cortical domain with trabecular atrophy suggests that it has a proactive or reactive role in determining bone mass and strength by facilitating musculoskeletal exchange in a form that is disengaged by ovariectomy.

Age Factors↗

PubMed1991

Direct Photolabeling with [P]UDP-Glucose for Identification of a Subunit of Cotton Fiber Callose Synthase.

We have identified a 52 kilodalton polypeptide as being a likely candidate for the catalytic subunit of the UDP-glucose: (1-->3)-beta-glucan (callose) synthase of developing fibers of Gossypium hirsutum (cotton). Such a polypeptide migrates coincident with callose synthase during glycerol gradient centrifugation in the presence of EDTA, and can be directly photolabeled with the radioactive substrate, alpha-[(32)P]UDP-glucose. Interaction with the labeled probe requires Ca(2+), a specific activator of callose synthase which is known to lower the K(m) of higher plant callose synthases for the substrate UDP-glucose. Using this probe and several other related ones, several other proteins which interact with UDP-glucose were also identified, but none satisfied all of the above criteria for being components of the callose synthase.

Journal Article↗

PubMed1993

Transient expression of a slow-tonic MHC isoform by extrafusal fibers in the developing rat.

ALD 19, a monoclonal antibody that recognizes the slow-tonic myosin heavy chain (MHC) isoform, has been used extensively as a marker for nuclear bag intrafusal fibers of muscle spindles in developing and adult rats. Extrafusal fibers of adult rat hindlimb muscles do not express slow-tonic MHC. However, while using ALD 19 to trace the fate of intrafusal fibers following neonatal denervation, we noted that some extrafusal fibers of neonates also bound this antibody. The immunolabeled extrafusal fibers were a subset of slow fibers located in the deep axial regions of crural muscles. The same fiber subset transiently displayed a weak affinity for ALD 19 during the first postnatal week in normal muscles. Denervation at birth increased the intensity of ALD 19 immunolabelling by these extrafusal fibers and extended the duration of the slow-tonic immunoreactivity into the 2nd postnatal week, after which expression diminished or ceased. Demonstration that some developing extrafusal fibers have a nerve-independent capacity for transiently expressing slow-tonic MHC, an MHC previously though to be expressed only by intrafusal fibers, raises the possibility that both types of fiber originate from a subset of bipotential slow primary myotubes in rat hindlimbs.

Animals↗