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Degeneration and regeneration of ventral root motor fibers in amyotrophic lateral sclerosis. Morphometric studies of cervical ventral roots.

Morphometric studies by histological and teased-fiber methods on myelinated fibers of the C6 and C8 ventral roots were made in cases of myotrophic lateral sclerosis (ALS) and in age-matched controls. In 11 controls myelinated fibers were divided into two groups: large and small fibers. The fiber density was calculated and the actual number of fibers was counted in both groups. The number of large fibers was significantly decreased in 14 of 17 cases with ALS, and correlated with the muscle strength of the ipsilateral upper limb as determined by manual muscle testing before death. The absolute number of small fibers of the C6 ventral root was increased in 3 of 5 cases of ALS. In these cases, there was a group of small myelinated fibers showing very thin myelin lamellae compared to axon circumference, and these were considered as regenerating myelinated fibers. On teased-fiber study, the frequency of fibers with axonal degeneration was 23.8% in ALS in contrast to 0.6% in controls (P less than 0.001). In addition, 5.5% of fibers showed segmental demyelination, and remyelination after segmental demyelination was found in 16.4% of fibers in ALS. These figures were also significantly larger (P less than 0.01) than controls (2.4% and 10.1%, respectively). These results suggest that the essential pathological change in the ventral spinal root in ALS is axonal degeneration of large myelinated fibers, and that Schwann cell involvement may also occur.

Adult↗

Effects of root replantation and neurotrophic factor treatment on long-term motoneuron survival and axonal regeneration after C7 spinal root avulsion.

In order to determine the effect of nerve root replantation on motoneuron survival and regeneration, we have avulsed and replanted C7 ventral rootlets in adult rabbits under various conditions. Intraspinal alterations and exact positions of ventrolateral replantations were studied in each animal, and the effects of BDNF and/or CNTF administration during replantation investigated in different experimental groups. Six months after lesion, about 70% of motoneurons were lost on the lesioned sides in the C7 segment, without significant differences between groups. Retrograde fluorescent tracing and histological analysis documented that many axons had regrown through the original ventral exit zones or had exited the spinal cord at the lateral replantation site. However, many laterally exiting axons had not grown out directly from the ventral horn through the lateral white matter but had elongated vertically before leaving the spinal cord. The mean axonal diameter was significantly higher in regenerated axons that had exited through the original ventral exit zones in comparison with axons which had grown out laterally. Application of BDNF and/or CNTF did not show any effects on the pathways of regeneration into the replanted root. The results indicate that motoneuron survival cannot be significantly improved by a single dose of neurotrophic factors applied to a ventrolateral replantation site. However, a significant number of myelinating axons are found in replanted roots, and regeneration may be more efficient when outgrowth through the original ventral exit zone is supported.

Animals↗

Cytokinin inhibits lateral root initiation but stimulates lateral root elongation in rice (Oryza sativa).

Research in lateral root (LR) development mainly focuses on the role of auxin. This article reports the effect of cytokinins (kinetin and trans-zeatin) on LR formation in rice (Oryza sativa L.). Our results showed that cytokinin has an inhibitory effect on LR initiation and stimulatory effect on LR elongation. Both KIN and ZEA at a concentration of 1 microM and above completely inhibited lateral root primordium (LRP) formation. The inhibitory effect of cytokinin on LR initiation required a continuous presence of KIN or ZEA in the growth solution. Cytokinin did not show any inhibitory effect on LR emergence from the seminal root once LRPs had been formed. The LRPs that developed in cytokinin-free solution can emerge normally in the solution containing inhibitory concentration (1 microM) of KIN and ZEA. The KIN and ZEA treatment dramatically stimulated LR elongation at all the concentrations tested. Maximum LR elongation was observed at a concentration of 0.01 microM KIN and 0.001 microM ZEA. The epidermal cell length increased significantly in LRs of cytokinin treated seedlings compared to those of untreated control. This result indicates that the stimulation of LR elongation by cytokinin is due to increased cell length. Exogenously applied auxin counteracted the effect of cytokinin on LR initiation and LR elongation, suggesting that cytokinin acts on LR elongation through an auxin dependent pathway.

Adenine↗

The SHORT-ROOT gene controls radial patterning of the Arabidopsis root through radial signaling.

Asymmetric cell divisions play an important role in the establishment and propagation of the cellular pattern of plant tissues. The SHORT-ROOT (SHR) gene is required for the asymmetric cell division responsible for formation of ground tissue (endodermis and cortex) as well as specification of endodermis in the Arabidopsis root. We show that SHR encodes a putative transcription factor with homology to SCARECROW (SCR). From analyses of gene expression and cell identity in genetically stable and unstable alleles of shr, we conclude that SHR functions upstream of SCR and participates in a radial signaling pathway. Consistent with a regulatory role in radial patterning, ectopic expression of SHR results in supernumerary cell divisions and abnormal cell specification in the root meristem.

Amino Acid Sequence↗

Some physiological and pharmacological properties of slow depolarization of substantia gelatinosa neurons by repetitive stimulation of C-fibers of dorsal root in adult rat spinal cord slices with dorsal root attached.

Transverse slices of the spinal cord with dorsal root attached were prepared from adult rats and used to record the response of the spinal substantia gelatinosa neurons evoked by repetitive stimulation of the dorsal root with a whole cell patch clamp method. The repetitive stimulation of 10 trains (20 pulses at 100 Hz/train) with an intensity necessary to activate C-fibers, but not A-fibers alone, evoked slow depolarization (SD) during it and the SD disappeared within 1 min after the termination of it. The SD was completely inhibited by 500 nM of tetrodotoxin (TTX), but not significantly changed by 50 nM of TTX, nor 20 microM CNQX + 50 microM AP5, nor 1 microM CP-99994. DAMGO inhibited the SD in a concentration dependent manner (10 nM-1 microM), but 1 microM DPDPE and 1 microM U-50488H did not. The inhibitory effect of DAMGO (1 microM) was reversed by naloxone (1 microM). These results suggest that the SD of substantia gelatinosa neurons evoked by repetitive stimulation of C-fibers of the dorsal root is an event relevant to nociception in the spinal dorsal horn.

2-Amino-5-phosphonovalerate↗

Pathogenesis of sciatic pain: role of herniated nucleus pulposus and deformation of spinal nerve root and dorsal root ganglion.

The basic pathophysiologic mechanisms related to disc herniation and sciatica are poorly understood. Recently it was demonstrated that nucleus pulposus from an intervertebral disc could induce structural and functional changes in adjacent nerve roots when applied epidurally, however, it is not known if such changes are painful. In a model for inducing disc herniation in the rat, we found that puncture of a lumbar disc with subsequent herniation of nucleus pulposus without nerve root compression, or chronic displacement of the 4th lumbar nerve root and ganglion, did not individually induce significant changes in thresholds for mechanical or thermal stimulation compared to sham-operated animals. However, the combination of disc puncture and displacement induced a reduction of the threshold for thermal stimulation, indicating hyperalgesia, that was present 2 days after surgery and gradually recovered during a 14-day period. These data and the associated description of this new model for experimental disc herniation may increase our understanding of the pathophysiologic events leading to sciatica and help in evaluating new modalities for diagnosis and treatment of disc herniation and sciatica.

Animals↗

Development of the root pole and cell patterning in Arabidopsis roots.

The root forms at the basal end of an axis that is set up early in embryogenesis, and recent genetic analysis has indicated that auxin transport is required for the formation of the root pole. Drug studies show that auxin transport is also required for the maintenance of the tissue organisation in the seedling root. These studies support existing models for tissue patterning that involve canalised auxin flow. Molecular insights into the mechanism of cell-type specification and patterning has come from the epidermis where positionally controlled cell specification is regulated by a cascade of transcription factors.

Arabidopsis↗

Root patterning: SHORT ROOT on the move.

The radial pattern of the plant root is determined by the action of two transcription factors, SHORT ROOT and SCARECROW, which are produced in different cell types. The SHORT ROOT protein has now been shown to move from cell to cell, regulating transcription of the downstream SCARECROW gene in the target cells.

Cell Division↗

Radioisotope studies of root activity and root-level interactions in tree-based production systems: a review.

This review focuses on the radioisotope methods used for studying the root activity patterns of woody perennials and root competition in multi-species cropping systems. 32P soil-injection is by far the most widely used method. The method is unique as it enables one to delineate the lateral and vertical spread of the absorbing roots. A system of classification of crop plants based on the concept of 'effective foraging space' has been developed. The scope and applicability of the radioisotope methods in different cropping situations as well as from the point of view of radiological safety were discussed.

Phosphorus Radioisotopes↗

Nonsurgical root canal therapy treatment with apparent indications for root-end surgery.

The recent introduction of the surgical microscope to the practice of endodontics, especially for surgery, has allowed clearer visualization of the periapex during root-end resection and filling. However, despite this and other technologic advances, it has not been demonstrated that in the absence of thorough canal debridement the success rate of periapical surgery has improved over the 50% to 60% demonstrated in most long-term prognosis studies. Therefore it remains important to fully instrument and obturate the root canal system with conventional therapy before surgery is considered; this considerably improves the long-term prognosis. Each of the case reports in this article involves a situation in which conventional treatment was performed when a surgical approach might have been considered as the treatment of choice. Surgery should not be considered to be the primary treatment when root canal treatment or retreatment may be readily achieved. Indeed, the operating microscope and other technical aids will probably allow more cases to be treated and retreated conventionally that might otherwise have required surgical intervention.

Adolescent↗

Comparison of radiographic and MRI features of a root-diverging odontogenic myxoma, with discussion of the differential diagnosis of lesions likely to move roots.

Lesions that can produce divergence of the roots of teeth in the mandible include odontogenic cysts (odontogenic keratocysts, lateral periodontal cysts and radicular cysts), ameloblastomas, odontogenic myxomas, central giant cell granulomas, adenomatoid odontogenic tumors and aneurismal bone cysts, and other tumors. Moreover most benign jaw lesions can do this occasionally. However, when lesions--which show interradicular tear-shaped radiolucencies--are small it is often difficult to interpret them radiographically, because they do not show characteristic radiographic features. We describe a comparison of radiographic and magnetic resonance (MR) features of a root-diverging odontogenic myxoma, with discussion of the differential diagnosis of lesions likely to move roots. In addition, we discuss radiographic and MR features of possible lesions, which show similar radiographic findings to odontogenic myxoma.

Adult↗

Expression of MsPG3-GFP fusions in Medicago truncatula'hairy roots' reveals preferential tip localization of the protein in root hairs.

Tip growth is a specialized type of polar growth where new cell wall is deposited in a localized region of the cell, the growing tip. These cells show a characteristic zonation, with a high accumulation of secretory vesicles containing cell wall components at the tip, followed by an organelle-enriched zone. MsPG3 is a Medicago sativa polygalacturonase gene isolated in our laboratory, specifically expressed during the interaction of this plant with its symbiotic partner Sinorhizobium meliloti and which might participate in tip growth processes during symbiosis. We have used MsPG3-GFP fusions to study in vivo protein transport processes and localization during root hair growth. Different MsPG3-GFP fusions were expressed in Medicago truncatula'hairy roots' following a protocol developed for this study and also tested by transient expression in onion epidermal cells. Preferential accumulation of an MsPG3-GFP fusion protein in the tip of the growing root hair at different developmental stages was found, confirming the delivery of MsPG3 to the newly synthesized cell wall. This indicates that this protein may participate in tip growth processes during symbiosis and, in addition, that this fusion could be a useful tool to study this process in plants.

Gene Transfer Techniques↗

AUX1 regulates root gravitropism in Arabidopsis by facilitating auxin uptake within root apical tissues.

Plants employ a specialized transport system composed of separate influx and efflux carriers to mobilize the plant hormone auxin between its site(s) of synthesis and action. Mutations within the permease-like AUX1 protein significantly reduce the rate of carrier-mediated auxin uptake within Arabidopsis roots, conferring an agravitropic phenotype. We are able to bypass the defect within auxin uptake and restore the gravitropic root phenotype of aux1 by growing mutant seedlings in the presence of the membrane-permeable synthetic auxin, 1-naphthaleneacetic acid. We illustrate that AUX1 expression overlaps that previously described for the auxin efflux carrier, AtPIN2, using transgenic lines expressing an AUX1 promoter::uidA (GUS) gene. Finally, we demonstrate that AUX1 regulates gravitropic curvature by acting in unison with the auxin efflux carrier to co-ordinate the localized redistribution of auxin within the Arabidopsis root apex. Our results provide the first example of a developmental role for the auxin influx carrier within higher plants and supply new insight into the molecular basis of gravitropic signalling.

Arabidopsis↗

Evolution and genetics of root hair stripes in the root epidermis.

Root hair pattern develops in a number of different ways in angiosperm. Cells in the epidermis of some species undergo asymmetric cell divisions to form a smaller daughter cell from which a hair grows, and a larger cell that forms a non-hair epidermal cell. In other species any cell in the epidermis can form a root hair. Hair cells are arranged in files along the Arabidopsis root, located in the gaps between underlying cortical cell files. Epidermal cells overlying a single cortical cell file develop as non-hair epidermal cells. Genetic analysis has identified a transcription factor cascade required for the formation of this pattern. WEREWOLF (WER) and GLABRA2 (GL2) are required for the formation of non-hair epidermal cells while CAPRICE (CPC) is required for hair cell development. Recent analyses of the pattern of epidermal cells among the angiosperms indicate that this striped pattern of cell organization evolved from non-striped ancestors independently in a number of diverse evolutionary lineages. The genetic basis for the evolution of epidermal pattern in angiosperms may now be examined.

Biological Evolution↗

Root, root hair, and symbiotic mutants of the model legume Lotus japonicus.

To gain an overview of plant factors controlling nodule number and organogenesis, an extensive screening using model legume Lotus japonicus was carried out. This screening involved 40,000 M2 seeds, and 32 stable mutant lines were isolated. From these, 16 mutant lines maintaining the phenotypic variation were selected and genetically analyzed. With respect to nodule number, four loci were identified, Ljsym77, Ljsym78, slippery root (slp), and radial organization1 (rdo1). The former two mutants have an increased number of nodules, while the latter two have a decreased number. Ljsym78-1 and Ljsym78-2 are hypernodulating mutants with a branched root system and were found to be allelic to Ljsym16. The phenotype of the Ljsym77 mutant was highly pleiotropic, being deficient in light and gravity responses. The slp mutant was isolated as a low-nodulating mutant lacking root hairs. Concerning nodule organogenesis, nine symbiotic loci were identified, including the two loci alb1 and fen1. Mutants affecting the developmental process of nodule organogenesis were placed in three phenotypic categories: Nod- (Ljsym70 to Ljsym73), Hist- (alb1-1, alb1-2, and Ljsym79), and Fix- (fen1, Ljsym75, and Ljsym81).

Ethyl Methanesulfonate↗

Resolution of furcation bone loss associated with vital pulp tissue after nonsurgical root canal treatment of three-rooted mandibular molars: a case report of identical twins.

This case report demonstrates the simultaneous development of furcation bone loss in three-rooted tooth #30 with vital pulp tissue in identical twins. In the first report, resolution of furcal bone loss was seen after nonsurgical root canal treatment. In the second report, furcal bone loss resolved after non-surgical root canal treatment, and periodontal therapy reduced probing depths to maintainable levels.

Adolescent↗

Changes in cytosolic pH within Arabidopsis root columella cells play a key role in the early signaling pathway for root gravitropism.

Ratiometric wide-field fluorescence microscopy with 1',7'- bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF)-dextran demonstrated that gravistimulation leads to rapid changes in cytoplasmic pH (pHc) in columella cells of Arabidopsis roots. The pHc of unstimulated columella cells in tiers 2 and 3, known sites of graviperception (E.B. Blancaflor, J.B. Fasano, S. Gilroy [1998] Plant Physiol 116: 213-222), was 7.22 +/- 0.02 pH units. Following gravistimulation, the magnitude and direction of pHc changes in these cells depended on their location in the columella. Cells in the lower side of tier 2 became more alkaline by 0.4 unit within 55 s of gravistimulation, whereas alkalinization of the cells on the upper side was slower (100 s). In contrast, all cells in tier 3 acidified by 0.4 pH unit within 480 s after gravistimulation. Disrupting these pHc changes in the columella cells using pHc modifiers at concentrations that do not affect root growth altered the gravitropic response. Acidifying agents, including bafilomycin A1, enhanced curvature, whereas alkalinizing agents disrupted gravitropic bending. These results imply that pHc changes in the gravisensing cells and the resultant pH gradients across the root cap are important at an early stage in the signal cascade leading to the gravitropic response.

Arabidopsis↗

Tissue-specific expression of stabilized SOLITARY-ROOT/IAA14 alters lateral root development in Arabidopsis.

Auxin is important for lateral root (LR) initiation and subsequent LR primordium development. However, the roles of tissue-specific auxin signaling in these processes are poorly understood. We analyzed transgenic Arabidopsis plants expressing the stabilized mutant INDOLE-3 ACETIC ACID 14 (IAA14)/SOLITARY-ROOT (mIAA14) protein as a repressor of the auxin response factors (ARFs), under the control of tissue-specific promoters. We showed that plants expressing the mIAA14-glucocorticoid receptor (GR) fusion protein under the control of the native IAA14 promoter had the solitary-root/iaa14 mutant phenotypes, including the lack of LR formation under dexamethasone (Dex) treatment, indicating that mIAA14-GR is functional in the presence of Dex. We then demonstrated that expression of mIAA14-GR under the control of the stele-specific SHORT-ROOT promoter suppressed LR formation, and showed that mIAA14-GR expression in the protoxylem-adjacent pericycle also blocked LR formation, indicating that the normal auxin response mediated by auxin/indole-3 acetic acid (Aux/IAA) signaling in the protoxylem pericycle is necessary for LR formation. In addition, we demonstrated that expression of mIAA14-GR under either the ARF7 or the ARF19 promoter also suppressed LR formation as in the arf7 arf19 double mutants, and that IAA14 interacted with ARF7 and ARF19 in yeasts. These results strongly suggest that mIAA14-GR directly inactivates ARF7/ARF19 functions, thereby blocking LR formation. Post-embryonic expression of mIAA14-GR under the SCARECROW promoter, which is expressed in the specific cell lineage during LR primordium formation, caused disorganized LR development. This indicates that normal auxin signaling in LR primordia, which involves the unknown ARFs and Aux/IAAs, is necessary for the establishment of LR primordium organization. Thus, our data show that tissue-specific expression of a stabilized Aux/IAA protein allows analysis of tissue-specific auxin responses in LR development by inactivating ARF functions.

Arabidopsis↗