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Involvement of leukemia inhibitory factor and nerve growth factor in the development of dorsal root ganglion neurons.

Leukemia inhibitory factor (LIF) was recently shown to stimulate the generation of sensory neurons from the murine neural crest in vitro. Here, we examine the respective activities of LIF and nerve growth factor (NGF) throughout the embryonic development of sensory neurons in dorsal root ganglia (DRG) and neural crest. In cultures of embryonic day 12 (E12) DRG, which contain sensory neuron precursor cells, a combination of both LIF and NGF are required for the differentiation of mature sensory neurons from their neurofilament negative (NF-) precursors. The primary differentiation step from NF- cell to NF+ immature neuron is promoted by LIF, whereas the survival and further maturation of the newly differentiated neurons depends on NGF. In cultures of sensory neurons isolated at the time of target innervation (E14 and E15 DRG), the survival of the majority of the neurons is dependent on NGF. However, LIF acts as a survival agent for a discrete population of NGF non-responsive neurons. From E16, the number of neurons maintained by LIF increases to > 90% by birth. Consistent with the in vitro observations, LIF mRNA could be detected at early developmental stages (E12-E13), within the spinal column and DRG as well as the limbs and, later (after E15), in areas of sensory innervation (skin, limbs, feet and gut). This supports the idea that LIF, as well as NGF, may regulate sensory development in vivo.

Animals↗

Rooting and the development of male sexual behavior in two inbred mouse strains.

The sexual development of C57BL 6/J and SEC/IReJ male mice was studied during twice-weekly pairings with estrus females from 5 weeks of age. SEC were far slower to begin copulating than C57 and their responses to the female included a much higher frequency of rooting. Rooting, where the male's snout is pushed below the female's flank and jerked upwards, declines over tests as the full copulatory pattern develops. SEC also spent a higher proportion of their time in close contact with the female and groomed her more than C57. Both strains showed more rapid development of copulation if testing was delayed until 10 weeks of age, especially if they had exposure to female scent during this maturation period. The frequency of rooting and the speed of development are inversely related in these strains, but in segregating hybrid populations no such relationship is maintained. It is argued that the slowness of SEC's development cannot be ascribed to low sexual motivation. The significance of rooting remains obscure, but it seems possible that it may represent an aspect of exploratory behavior that is highly developed in SEC and some other strains.

Animals↗

Adhesion and development of the root rot fungus (Heterobasidion annosum) on conifer tissues: effects of spore and host surface constituents.

The objective of this study was to correlate the occurrence of particular root and woody stump surface components with the ability of spores of the root rot fungus (Heterobasidion annosum) to adhere, germinate and establish on conifer tissues. With the aid of high performance liquid chromatography, several sugars (pinitol, xylitol, dulcitol, mannitol, D-glucose, mannose, fructose) were detected on both stump and fine root surfaces of Scots pine and Norway spruce. Of all the sugars observed, xylose and arabinose were poorly utilized for initiation of germ tube growth whereas spore germination was enhanced in the presence of D-glucose, mannose or fructose. Oxidation of these sugars by pretreatment of wood discs or roots with periodic acid abolished the ability of the spores to germinate. Non-sugar components such as long chain fatty acids on spores and root surfaces as detected with nuclear magnetic resonance were found to have a significant influence on adhesion and initiation of germ tube development. Removal of these aliphatic compounds from the root surface increased spore germination by 2-fold, whereas similar treatment on spores led to a 5-fold decrease in adhesiveness to root material. In vitro studies revealed that the di-ethyl ether extract from the roots had no long term adverse effect on spore germination which suggests that the fungus may possess the capability to detoxify this substance. Similarly, adhesion of spores was affected by low and freezing temperatures. The role of significant levels of mannitol and trehalose accumulated in spores and hyphae of the fungi on viability, survival and tolerance to adverse conditions such as oxidative stress, freezing and desiccation are discussed.

Journal Article↗

The consequence of peroxidase overexpression in transgenic plants on root growth and development.

Transgenic tobacco plants that overproduce the tobacco anionic peroxidase wilt upon reaching maturity, although having functional stomata and normal vascular anatomy and physiology. These plants were examined further to determine the cause for wilting, and thus better understand how the anionic peroxidase functions in plant growth and development. Shoots from young peroxidase overproducing plants were grafted onto wild-type tobacco root stock to determine if the roots could absorb and transmit sufficient water to maintain leaf turgidity. These grafted plants never wilted when grown in the greenhouse though shoot peroxidase activity remained ten-fold greater than in control plants, thus indicating that wilting is a consequence of peroxidase expression in the roots. Close examination of root systems revealed considerably less root mass in the transformed plant, primarily exhibited through a decrease in branching. At flowering, root growth rate and total root mass in transformed plants were less than 50% of control plants although shoot mass and growth rate were unchanged. This is in contrast to root growth in young seedlings where transformed plants performed equivalently to controls. Root hydraulic conductivity was measured to evaluate the effect of elevated peroxidase expression on water absorption and transport; however, no significant change in hydraulic conductivity was found in transformed plants. The consequence of anionic peroxidase overexpression on indoleacetic acid (IAA) metabolism was also examined. No significant difference in IAA levels was observed; however, root elongation in plants overexpressing peroxidase was insensitive to exogenous IAA. It can be concluded that the overexpression of the tobacco anionic peroxidase in transformed plants results in diminished root mass from fewer root branches, which contributes to the wilting phenomenon seen in these plants. Further, this developmental change in transformed plants may be a consequence of the metabolism of IAA by the anionic peroxidase.

Indoleacetic Acids↗

LRP1, a gene expressed in lateral and adventitious root primordia of arabidopsis.

We describe a gene that is expressed in lateral and adventitious root primordia of Arabidopsis. The gene was identified by expression of a transposon-borne promoterless beta-glucuronidase gene in lateral root primordia. The gene, designated LRP1 for lateral root primordium 1, and its corresponding cDNA were cloned and sequenced. The expression pattern of the gene in lateral root primordia was confirmed by in situ hybridization with LRP1 cDNA probes. The LRP1 gene encodes a novel protein. LRP1 expression is activated during the early stages of root primordium development and is turned off prior to the emergence of lateral roots from the parent root. Insertion of the transposon in the LRP1 gene disrupted its expression. To evaluate the homozygous insertion line for a mutant phenotype, several aspects of wild-type lateral root development were analyzed. A mutant phenotype has not yet been identified in the insertion line; however, there is evidence that the gene belongs to a small gene family. LRP1 provides a molecular marker to study the early stages of lateral and adventitious root primordium development.

Amino Acid Sequence↗

Pscroph, a parasitic plant EST database enriched for parasite associated transcripts.

BACKGROUND: Parasitic plants in the Orobanchaceae develop invasive root haustoria upon contact with host roots or root factors. The development of haustoria can be visually monitored and is rapid, highly synchronous, and strongly dependent on host factor exposure; therefore it provides a tractable system for studying chemical communications between roots of different plants. DESCRIPTION: Triphysaria is a facultative parasitic plant that initiates haustorium development within minutes after contact with host plant roots, root exudates, or purified haustorium-inducing phenolics. In order to identify genes associated with host root identification and early haustorium development, we sequenced suppression subtractive libraries (SSH) enriched for transcripts regulated in Triphysaria roots within five hours of exposure to Arabidopsis roots or the purified haustorium-inducing factor 2,6 dimethoxybenzoquinone. The sequences of over nine thousand ESTs from three SSH libraries and their subsequent assemblies are available at the Pscroph database http://pscroph.ucdavis.edu. The web site also provides BLAST functions and allows keyword searches of functional annotations. CONCLUSION: Libraries prepared from Triphysaria roots treated with host roots or haustorium inducing factors were enriched for transcripts predicted to function in stress responses, electron transport or protein metabolism. In addition to parasitic plant investigations, the Pscroph database provides a useful resource for investigations in rhizosphere interactions, chemical signaling between organisms, and plant development and evolution.

Arabidopsis↗

An ultrastructural study of cytodifferentiation in the developing human root-tip.

Transmission electron microscopy showed that in growing human teeth, the root sheath consisted of inner and outer epithelial cells. The inner epithelial cells formed a basal lamina associated filamentous layer which increased in density in coronal direction. Extensions of developing odontoblasts were in contact with the basal lamina. Direct contact with the epithelial plasmalemma was not observed. The odontoblasts obtained their fully-developed cylindrical appearance after making contact with the basal lamina of the inner epithelial cells. But, they possessed already abundant RER before these contacts were present, indicating that the differentiation of dental papillary cells into collagen-producing cells did not require heterotypic epithelio-mesenchymal contacts. After deposition of dentine, the odontoblast processes were withdrawn from the epithelium and the outer layer of root dentine. Differentiation of cementoblasts was observed in the dental follicle in which mesenchymal cells develop into RER-containing cells, migrating through apical root-sheath fenestrations to their final position between the continuous root sheath and outer dentinal layer. The findings suggest that all differentiation steps of odontogenic epithelium and odontogenic papillary and follicular mesenchyme proceed without actual contact between epithelium and mesenchyme.

Bicuspid↗

Evidence for a rostrocaudal organization in dorsal root ganglia during development as demonstrated by intra-uterine WGA-HRP injections into the hindlimb of rat fetuses.

The development of the sensory innervation of the rat hindlimb was studied with special attention to the dorsal root ganglia and the lumbar plexus. Injections of wheat germ agglutinin-horseradish peroxidase were made into the hindlimb of 30 rat fetuses of gestational ages ranging from embryonic day 15-18. Additionally wheat germ agglutinin-horseradish peroxidase was applied to the sciatic nerves of 8 neonatal rats and 3 adults. The saphenous nerves of 2 neonatal rats were labeled. Injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the hindlimb of the fetuses result in completely and partially labeled dorsal root ganglia. Partial labeling always concerns the rostral or caudal part of a dorsal root ganglion. The associated dorsal roots of partially labeled dorsal root ganglia are also partially labeled in a corresponding rostrocaudal fashion. Reconstructions of the labeled nerves following injections into the hindlimb suggested that the somata of the sensory neurons of a particular nerve can be restricted to the rostral or caudal half of a dorsal root ganglion. For example: the rostral half of the fourth lumbar dorsal root ganglion belongs to the femoral nerve and its caudal half to the sciatic nerve. The results of application of wheat germ agglutinin-horseradish peroxidase to the central ends of the cut sciatic and saphenous nerves in neonatal rats confirmed these observations. So the rostrocaudal organization in the dorsal root ganglia stems from the distribution pattern of the peripheral nerves.

Animals↗

Cell-type-specific expression of plant cytochrome c mRNA in developing flowers and roots.

We have used RNA in situ hybridization to analyze the expression of transcripts encoding cytochrome c in different tissues and organs of sunflower (Helianthus annuus). Although northern-blot hybridization experiments indicate that the relative abundance of transcripts does not vary greatly, we have detected important changes in localization during flower development. Enhanced expression is observed in floral meristems as soon as they are discernible from the central portion of the capitulum containing the inflorescence meristem. As flowers develop, labeling is observed in all developing floral organ primordia. Later in development, expression in petals is reduced, and only the central portion of the flower becomes labeled. During the process of stamen formation, hybridization signals were obtained mainly in anthers. Less developed flowers at this stage showed expression through the archesporial tissue. During meiosis, the label was observed mainly in tapetal cells. Specific expression patterns, similar to those obtained for sunflower, were observed when Arabidopsis flowers were analyzed with a homologous cytochrome c probe. Specific patterns of expression were also observed in young sunflower roots. In this case, enhanced expression was detected in developing endodermis and pericycle and in protoxylem initials. We conclude that cell-specific mechanisms operate to regulate the abundance of cytochrome c encoding transcripts in different plant tissues. The overlap between the expression patterns of the nuclear encoded cytochrome c gene and some mitochondrial genes suggests the existence of coordinated mechanisms of expression.

Cytochrome c Group↗

Role of cytokinin and auxin in shaping root architecture: regulating vascular differentiation, lateral root initiation, root apical dominance and root gravitropism.

BACKGROUND AND AIMS: Development and architecture of plant roots are regulated by phytohormones. Cytokinin (CK), synthesized in the root cap, promotes cytokinesis, vascular cambium sensitivity, vascular differentiation and root apical dominance. Auxin (indole-3-acetic acid, IAA), produced in young shoot organs, promotes root development and induces vascular differentiation. Both IAA and CK regulate root gravitropism. The aims of this study were to analyse the hormonal mechanisms that induce the root's primary vascular system, explain how differentiating-protoxylem vessels promote lateral root initiation, propose the concept of CK-dependent root apical dominance, and visualize the CK and IAA regulation of root gravitropiosm. KEY ISSUES: The hormonal analysis and proposed mechanisms yield new insights and extend previous concepts: how the radial pattern of the root protoxylem vs. protophloem strands is induced by alternating polar streams of high IAA vs. low IAA concentrations, respectively; how differentiating-protoxylem vessel elements stimulate lateral root initiation by auxin-ethylene-auxin signalling; and how root apical dominance is regulated by the root-cap-synthesized CK, which gives priority to the primary root in competition with its own lateral roots. CONCLUSIONS: CK and IAA are key hormones that regulate root development, its vascular differentiation and root gravitropism; these two hormones, together with ethylene, regulate lateral root initiation.

Cytokinins↗

Pseudoaneurysm developed after aortic root homograft implantation.

A 55-year-old man with severe infective endocarditis underwent aortic root replacement using a homograft concomitant with saphenous vein grafting to the left anterior descending artery. The patient developed angina due to stenosis of the proximal anastomosis of the vein graft accompanied by a 2-cm pseudoaneurysm. This complex lesion was successfully managed with urgent coronary artery bypass surgery through a left thoracotomy, followed by a redo replacement of the old aortic homograft. The mechanism and the basis of the treatment strategy adopted for this case are described.

Anastomosis, Surgical↗

Interaction between two auxin-resistant mutants and their effects on lateral root formation in rice (Oryza sativa L.).

Since root elongation is very sensitive to auxin, screening for reduced inhibition in root elongation has been an important method for the detection of auxin-resistant mutants. Two recessive auxin-resistant lines of rice (Oryza sativa L. ssp. indica cv. IR8), arm1 and arm2, have been isolated by screening for resistance to 2,4-dichlorophenoxyacetic acid (2,4-D). arm1 displays a variety of morphological defects including reduced lateral root formation, increased seminal root elongation, reduced root diameter, and impaired xylem development in roots, while the arm2 phenotype is almost similar to wild-type IR8 except for a slightly reduced lateral root formation, impaired xylem development in roots and an enhanced plant height. Although the growth of arm2 roots exhibited a resistance to 2,4-D, it was sensitive to 1-naphthaleneacetic acid (NAA) as the wild type. At the same time, the arm2 roots showed a reduced [14C]2,4-D uptake while uptake of [3H]NAA was normal, suggesting that the resistance to 2,4-D of arm2 roots is due to a defect in 2,4-D uptake. To investigate the possible interaction between arm1 and arm2 genes, a double mutant has been constructed. The roots of arm1 arm2 double mutant were more resistant to 2,4-D and formed fewer lateral roots than those of either single mutant, suggesting that the two genes show synergistic effects with respect to both auxin response and lateral root formation. By contrast, all these mutants displayed the normal gravitropic response in roots, as did the wild-type plants. Taken together, Arm1 and Arm2 genes seem to function in different processes in the auxin-response pathways leading to lateral root formation.

2,4-Dichlorophenoxyacetic Acid↗