RESUMEN
Plants maintain pools of pluripotent stem cells which allow them to constantly produce new tissues and organs. Stem cell homeostasis in shoot and root tips depends on negative regulation by ligand-receptor pairs of the CLE peptide and leucine-rich repeat receptor-like kinase (LRR-RLK) families. However, regulation of the cambium, the stem cell niche required for lateral growth of shoots and roots, is poorly characterized. Here we show that the LRR-RLK MOL1 is necessary for cambium homeostasis in Arabidopsis thaliana. By employing promoter reporter lines, we reveal that MOL1 is active in a domain that is distinct from the domain of the positively acting CLE41/PXY signaling module. In particular, we show that MOL1 acts in an opposing manner to the CLE41/PXY module and that changing the domain or level of MOL1 expression both result in disturbed cambium organization. Underlining discrete roles of MOL1 and PXY, both LRR-RLKs are not able to replace each other when their expression domains are interchanged. Furthermore, MOL1 but not PXY is able to rescue CLV1 deficiency in the shoot apical meristem. By identifying genes mis-expressed in mol1 mutants, we demonstrate that MOL1 represses genes associated with stress-related ethylene and jasmonic acid hormone signaling pathways which have known roles in coordinating lateral growth of the Arabidopsis stem. Our findings provide evidence that common regulatory mechanisms in different plant stem cell niches are adapted to specific niche anatomies and emphasize the importance of a complex spatial organization of intercellular signaling cascades for a strictly bidirectional tissue production.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/metabolismo , Cámbium/metabolismo , Proteínas Quinasas/fisiología , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Cámbium/crecimiento & desarrollo , Ciclopentanos/metabolismo , Etilenos/metabolismo , Regulación de la Expresión Génica de las Plantas , Meristema/crecimiento & desarrollo , Meristema/metabolismo , Mutación , Oxilipinas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismo , Transducción de SeñalRESUMEN
Transport of RNAs to dendrites occurs in neuronal RNA granules, which allows local synthesis of specific proteins at active synapses on demand, thereby contributing to learning and memory. To gain insight into the machinery controlling dendritic mRNA localization and translation, we established a stringent protocol to biochemically purify RNA granules from rat brain. Here, we identified a specific set of interactors for two RNA-binding proteins that are known components of neuronal RNA granules, Barentsz and Staufen2. First, neuronal RNA granules are much more heterogeneous than previously anticipated, sharing only a third of the identified proteins. Second, dendritically localized mRNAs, e.g., Arc and CaMKIIα, associate selectively with distinct RNA granules. Third, our work identifies a series of factors with known roles in RNA localization, translational control, and RNA quality control that are likely to keep localized transcripts in a translationally repressed state, often in distinct types of RNPs.
Asunto(s)
Dendritas/metabolismo , Biosíntesis de Proteínas , Transporte de ARN , ARN Mensajero/metabolismo , Animales , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/genética , Proteína Quinasa Tipo 2 Dependiente de Calcio Calmodulina/metabolismo , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Hipocampo/citología , Hipocampo/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Unión Proteica , Proteínas de Unión al ARN/metabolismo , Ratas , Ribonucleoproteínas/metabolismoRESUMEN
Unlike the situation in animals, the final morphology of the plant body is highly modulated by the environment. During Arabidopsis development, intrinsic factors provide the framework for basic patterning processes. CLASS III HOMEODOMAIN LEUCINE ZIPPER (HD-ZIPIII) transcription factors are involved in embryo, shoot and root patterning. During vegetative growth HD-ZIPIII proteins control several polarity set-up processes such as in leaves and the vascular system. We have identified several direct target genes of the HD-ZIPIII transcription factor REVOLUTA (REV) using a chromatin immunoprecipitation/DNA sequencing (ChIP-Seq) approach. This analysis revealed that REV acts upstream of auxin biosynthesis and affects directly the expression of several class II HD-ZIP transcription factors that have been shown to act in the shade-avoidance response pathway. We show that, as well as involvement in basic patterning, HD-ZIPIII transcription factors have a critical role in the control of the elongation growth that is induced when plants experience shade. Leaf polarity is established by the opposed actions of HD-ZIPIII and KANADI transcription factors. Finally, our study reveals that the module that consists of HD-ZIPIII/KANADI transcription factors controls shade growth antagonistically and that this antagonism is manifested in the opposed regulation of shared target genes.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Tipificación del Cuerpo , Proteínas de Homeodominio/genética , Factores de Transcripción/genética , Adaptación Fisiológica , Arabidopsis/citología , Arabidopsis/crecimiento & desarrollo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/metabolismo , Sitios de Unión , Inmunoprecipitación de Cromatina , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genoma de Planta/genética , Proteínas de Homeodominio/metabolismo , Hipocótilo/citología , Hipocótilo/genética , Hipocótilo/crecimiento & desarrollo , Hipocótilo/efectos de la radiación , Hibridación in Situ , Ácidos Indolacéticos/análisis , Ácidos Indolacéticos/metabolismo , Luz , Mutación , Filogenia , Hojas de la Planta/citología , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/efectos de la radiación , Análisis de Secuencia de ADN , Transducción de Señal , Factores de Transcripción/metabolismoRESUMEN
Long distance cell-to-cell communication is critical for the development of multicellular organisms. In this respect, plants are especially demanding as they constantly integrate environmental inputs to adjust growth processes to different conditions. One example is thickening of shoots and roots, also designated as secondary growth. Secondary growth is mediated by the vascular cambium, a stem cell-like tissue whose cell-proliferating activity is regulated over a long distance by the plant hormone auxin. How auxin signaling is integrated at the level of cambium cells and how cambium activity is coordinated with other growth processes are largely unknown. Here, we provide physiological, genetic, and pharmacological evidence that strigolactones (SLs), a group of plant hormones recently described to be involved in the repression of shoot branching, positively regulate cambial activity and that this function is conserved among species. We show that SL signaling in the vascular cambium itself is sufficient for cambium stimulation and that it interacts strongly with the auxin signaling pathway. Our results provide a model of how auxin-based long-distance signaling is translated into cambium activity and suggest that SLs act as general modulators of plant growth forms linking the control of shoot branching with the thickening of stems and roots.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , Lactonas/metabolismo , Transducción de Señal , Arabidopsis/efectos de los fármacos , Cámbium/citología , Cámbium/efectos de los fármacos , Cámbium/crecimiento & desarrollo , Inflorescencia/citología , Inflorescencia/genética , Inflorescencia/crecimiento & desarrollo , Lactonas/farmacología , Mutación/genética , Especificidad de Órganos/efectos de los fármacos , Tallos de la Planta/citología , Tallos de la Planta/efectos de los fármacos , Tallos de la Planta/crecimiento & desarrollo , Transducción de Señal/efectos de los fármacos , Especificidad de la EspecieRESUMEN
Multipotent stem cell populations, the meristems, are fundamental for the indeterminate growth of plant bodies. One of these meristems, the cambium, is responsible for extended root and stem thickening. Strikingly, although the pivotal role of the plant hormone auxin in promoting cambium activity has been known for decades, the molecular basis of auxin responsiveness on the level of cambium cells has so far been elusive. Here, we reveal that auxin-dependent cambium stimulation requires the homeobox transcription factor WOX4. In Arabidopsis thaliana inflorescence stems, 1-N-naphthylphthalamic acid-induced auxin accumulation stimulates cambium activity in the wild type but not in wox4 mutants, although basal cambium activity is not abolished. This conclusion is confirmed by the analysis of cellular markers and genome-wide transcriptional profiling, which revealed only a small overlap between WOX4-dependent and cambium-specific genes. Furthermore, the receptor-like kinase PXY is required for a stable auxin-dependent increase in WOX4 mRNA abundance and the stimulation of cambium activity, suggesting a concerted role of PXY and WOX4 in auxin-dependent cambium stimulation. Thus, in spite of large anatomical differences, our findings uncover parallels between the regulation of lateral and apical plant meristems by demonstrating the requirement for a WOX family member for auxin-dependent regulation of lateral plant growth.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Cámbium/citología , Proteínas de Homeodominio/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Cámbium/crecimiento & desarrollo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas de Homeodominio/genética , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/metabolismo , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismoRESUMEN
UNLABELLED: Diethylnitrosamine (DEN) is a hepatic procarcinogen which is frequently used as an inducer of hepatocellular carcinoma (HCC) in mice. Although mice after DEN exposure are among the most widely used models for liver tumorigenesis, a detailed, mechanistic characterization of the longitudinal changes in the respective tumor genomes has never been performed. Here we established the chronological order of genetic alterations during DEN carcinogenesis by examining mice at different points in time. Tumor samples were isolated by laser microdissection and subjected to array-comparative genomic hybridization (array-CGH) and sequencing analysis. Chromosomal gains and losses were observed in tumors by week 32 and increased significantly by week 56. Loss of distal chromosome 4q, including the tumor suppressors Runx3 and Nr0b2/Shp, was a frequent early event and persisted during all tumor stages. Surprisingly, sequencing revealed that ß-catenin mutations occurred late and were clearly preceded by chromosomal instability. Thus, contrary to common belief, ß-catenin mutations and activation of the Wnt/ß-catenin pathway are not involved in tumor initiation in this model of chemical hepatocarcinogenesis. CONCLUSION: Our study suggests that the majority of the current knowledge about genomic changes in HCC is based on advanced tumor lesions and that systematic analyses of the chronologic order including early lesions may reveal new, unexpected findings.
Asunto(s)
Inestabilidad Genómica , Animales , Carcinoma Hepatocelular/inducido químicamente , Carcinoma Hepatocelular/genética , Hibridación Genómica Comparativa , Subunidad alfa 3 del Factor de Unión al Sitio Principal/genética , Variaciones en el Número de Copia de ADN , Dietilnitrosamina , Humanos , Neoplasias Hepáticas/inducido químicamente , Neoplasias Hepáticas/genética , Masculino , Ratones , Receptores Citoplasmáticos y Nucleares/genética , beta Catenina/genéticaRESUMEN
Cell-to-cell communication is crucial for the development of multicellular organisms, especially during the generation of new tissues and organs. Secondary growth--the lateral expansion of plant growth axes--is a highly dynamic process that depends on the activity of the cambium. The cambium is a stem cell-like tissue whose activity is responsible for wood production and, thus, for the establishment of extended shoot and root systems. Attempts to study cambium regulation at the molecular level have been hampered by the limitations of performing genetic analyses in trees and by the difficulty of accessing this tissue in model systems such as Arabidopsis thaliana. Here, we describe the roles of two receptor-like kinases, REDUCED IN LATERAL GROWTH1 (RUL1) and MORE LATERAL GROWTH1 (MOL1), as opposing regulators of cambium activity. Their identification was facilitated by a novel in vitro system in which cambium formation is induced in isolated Arabidopsis stem fragments. By combining this system with laser capture microdissection, we characterized transcriptome remodeling in a tissue- and stage-specific manner and identified series of genes induced during different phases of cambium formation. In summary, we provide a means for investigating cambium regulation in unprecedented depth and present two signaling components that control a process responsible for the accumulation of a large proportion of terrestrial biomass.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/genética , Cámbium/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas Quinasas/metabolismo , Arabidopsis/citología , Proteínas de Arabidopsis/genética , Biomarcadores/metabolismo , Cámbium/citología , Cámbium/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Genes de Plantas/genética , Especificidad de Órganos/genética , Tallos de la Planta/genética , Tallos de la Planta/crecimiento & desarrollo , Proteínas Quinasas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
After primary growth, most dicotyledonous plants undergo secondary growth. Secondary growth involves an increase in the diameter of shoots and roots through formation of secondary vascular tissue. A hallmark of secondary growth initiation in shoots of dicotyledonous plants is the initiation of meristematic activity between primary vascular bundles, i.e. in the interfascicular regions. This results in establishment of a cylindrical meristem, namely the vascular cambium. Surprisingly, despite its major implications for plant growth and the accumulation of biomass, the molecular regulation of secondary growth is only poorly understood. Here, we combine histological, molecular and genetic approaches to characterize interfascicular cambium initiation in the Arabidopsis thaliana inflorescence shoot. Using genome-wide transcriptional profiling, we show that stress-related and touch-inducible genes are up-regulated in stem regions where secondary growth takes place. Furthermore, we show that the products of COI1, MYC2, JAZ7 and the touch-inducible gene JAZ10, which are components of the JA signalling pathway, are cambium regulators. The positive effect of JA application on cambium activity confirmed a stimulatory role of JA in secondary growth, and suggests that JA signalling triggers cell divisions in this particular context.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Cámbium/crecimiento & desarrollo , Ciclopentanos/farmacología , Oxilipinas/farmacología , Arabidopsis/anatomía & histología , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Cámbium/anatomía & histología , Cámbium/genética , División Celular/genética , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Meristema/anatomía & histología , Meristema/genética , Meristema/crecimiento & desarrollo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Reguladores del Crecimiento de las Plantas/farmacología , Raíces de Plantas/anatomía & histología , Raíces de Plantas/genética , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The dendritic localization of mRNAs and their subsequent translation at stimulated synapses contributes to the experience-dependent remodeling of synapses and thereby to the establishment of long-term memory. Localized mRNAs are transported in a translationally silent manner to distal dendrites in specific ribonucleoprotein particles (RNPs), termed transport RNPs. A recent study suggested that processing bodies (P-bodies), which have recently been identified as sites of RNA degradation and translational control in eukaryotic cells, may participate in the translational control of dendritically localized mRNAs in Drosophila neurons. This study raised the interesting question of whether dendritic transport RNPs are distinct from P-bodies or whether those structures share significant overlap in their molecular composition in mammalian neurons. Here, we show that P-body and transport RNP markers do not colocalize and are not transported together in the same particles in dendrites of mammalian neurons. Detailed time-lapse videomicroscopy analyses reveal, however, that both P-bodies and transport RNPs can interact in a dynamic manner via docking. Docking is a frequent event involving as much as 50% of all dendritic P-bodies. Chemically induced neuronal activity results in a 60% decrease in the number of P-bodies in dendrites, suggesting that P-bodies disassemble after synaptic stimulation. Our data lend support to the exciting hypothesis that dendritically localized mRNAs might be stored in P-bodies and be released and possibly translated when synapses become activated.
Asunto(s)
Dendritas/metabolismo , Hipocampo/citología , Neuronas/citología , Proteínas de Unión al ARN/metabolismo , Ribonucleoproteínas/metabolismo , Animales , Transporte Biológico/fisiología , Factor Neurotrófico Derivado del Encéfalo/farmacología , Células Cultivadas , Embrión de Mamíferos , Fármacos actuantes sobre Aminoácidos Excitadores/farmacología , Ácido Glutámico/farmacología , Proteínas Fluorescentes Verdes/metabolismo , Peróxido de Hidrógeno/farmacología , Indoles , Microscopía Confocal/métodos , Microscopía por Video/métodos , Ratas , Transfección/métodosRESUMEN
Background. The aim of the present study was to investigate the relationships between personality and quality of life during the course of geriatric rehabilitation, against the background of Cloninger's biosocial theory of personality. Methods. All consecutive patients of a geriatric rehabilitation clinic during one year were evaluated at admission and discharge (N = 687) by means of the ''Vienna List'' (a newly developed questionnaire for the assessment of quality of life in patients with severe dementia), and two variants of the Temperament and Character Inventory. Results. Self-directedness showed the most general and highest impact on quality of life and successful rehabilitation. Conclusions. It is probable in old and very old individuals who are on their highest level of maturity that the character represents the most important regulatory system in the encounter with challenges of daily life, which necessitates rehabilitation.