Molecular Aspects of Secretory Granule Exocytosis by Neurons and Endocrine Cells

Neuronal communication and endocrine signaling are fundamental for integrating the function of tissues and cells in the body. Hormones released by endocrine cells are transported to the target cells through the circulation. By contrast, transmitter release from neurons occurs at specialized intercellular junctions, the synapses. Nevertheless, the mechanisms by which signal molecules are synthesized, stored, and eventually secreted by neurons and endocrine cells are very similar. Neurons and endocrine cells have in common two different types of secretory organelles, indicating the presence of two distinct secretory pathways. The synaptic vesicles of neurons contain excitatory or inhibitory neurotransmitters, whereas the secretory granules (also referred to as dense core vesicles, because of their electron dense content) are filled with neuropeptides and amines. In endocrine cells, peptide hormones and amines predominate in secretory granules. The function and content of vesicles, which share antigens with synaptic vesicles, are unknown for most endocrine cells. However, in B cells of the pancreatic islet, these vesicles contain GABA, which may be involved in intrainsular signaling.' Exocytosis of both synaptic vesicles and secretory granules is controlled by cytoplasmic calcium. However, the precise mechanisms of the subsequent steps, such as docking of vesicles and fusion of their membranes with the plasma membrane, are still incompletely understood. This contribution summarizes recent observations that elucidate components in neurons and endocrine cells involved in exocytosis. Emphasis is put on the intracellular aspects of the release of secretory granules that recently have been analyzed in detail

Pareto versus lognormal: a maximum entropy test

It is commonly found that distributions that seem to be lognormal over a broad range change to a power-law (Pareto) distribution for the last few percentiles. The distributions of many physical, natural, and social events (earthquake size, species abundance, income and wealth, as well as file, city, and firm sizes) display this structure. We present a test for the occurrence of power-law tails in statistical distributions based on maximum entropy. This methodology allows one to identify the true data-generating processes even in the case when it is neither lognormal nor Pareto. The maximum entropy approach is then compared with other widely used methods and applied to different levels of aggregation of complex systems. Our results provide support for the theory that distributions with lognormal body and Pareto tail can be generated as mixtures of lognormally distributed units

Regional and developmental brain expression patterns of SNAP25 splice variants

SNAP25 is an essential SNARE protein for regulated exocytosis in neuronal cells. Differential splicing of the SNAP25 gene results in the expression of two transcripts, SNAP25a and SNAP25b. These splice variants differ by only 9 amino acids, and studies of their expression to date have been limited to analysis of the corresponding mRNAs. Although these studies have been highly informative, it is possible that factors such as differential turnover of the SNAP25 proteins could complicate interpretations based entirely on mRNA expression profiles

UBA1/GARS-dependent pathways drive sensory-motor connectivity defects in spinal muscular atrophy

Deafferentation of motor neurons as a result of defective sensory-motor connectivity is a critical early event in the pathogenesis of spinal muscular atrophy, but the underlying molecular pathways remain unknown. We show that restoration of ubiquitin-like modifier-activating enzyme 1 (UBA1) was sufficient to correct sensory-motor connectivity in the spinal cord of mice with spinal muscular atrophy. Aminoacyl-tRNA synthetases, including GARS, were identified as downstream targets of UBA1. Regulation of GARS by UBA1 occurred via a non-canonical pathway independent of ubiquitylation. Dysregulation of UBA1/GARS pathways in spinal muscular atrophy mice disrupted sensory neuron fate, phenocopying GARS-dependent defects associated with Charcot-Marie-Tooth disease. Sensory neuron fate was corrected following restoration of UBA1 expression and UBA1/GARS pathways in spinal muscular atrophy mice. We conclude that defective sensory motor connectivity in spinal muscular atrophy results from perturbations in a UBA1/GARS pathway that modulates sensory neuron fate, thereby highlighting significant molecular and phenotypic overlap between spinal muscular atrophy and Charcot-Marie-Tooth disease.</p

Soils with dark subsurface horizons in saline basins in the Brazilian Pantanal.

Soils in the Brazilian Pantanal classified as Espodossolos and registered in the literature diverge from the central concept of pedogenesis by podzolization, especially due to the high values of pH and basic cations, although the morphology is similar to that of spodic horizons. In this sense, this study hypothesized that the neutral to alkaline conditions along the edges of the saline basins in the Lower Nhecolândia region do not inhibit the podzolization process nor the development of spodic soils. The objectives were to analyze the attributes of soils with spodic features and evaluate whether they correspond to a podzolization process. Four profiles in a transect in Lower Nhecolândia were selected, where the morphology indicated the presence of a spodic horizon. Three profiles (P1, P2 and P4) were located in a bay and one in the transition between a bay and a "cordilheira" (small elevation between lagoons) (P3). The soils were analyzed for particle size, chemistry and X-ray fluorescence (XRF), as well as organic carbon (C org), total carbon (TC) and XRF of the nodules. The complexity of features and characteristics of soil profiles in the Lower Nhecolândia region indicate multiple pedogenetic processes in this environment. The characteristics of all profiles denote a redoximorphic process: poor profile development (except for P3), mottles formed by precipitation of Fe and Mn oxides, as well as presence of Fe and Mn nodules. The organic matter content of these nodules is mostly greater than or equal to that of the surrounding material, and aluminum is relatively not higher than in the fine earth. This reinforces a rexodimorphic process in the horizons with spodic features. The occurrence of sodic and solodic characters, as well as clay accumulation in P3 also characterize the sodification process. The neutral to alkaline pH values in water, high sum of bases and low C org and Al oxide contents of the studied soils all contradict the occurrence of a podzolization process. However, Fe, Al and C org accumulation in the nodules and some B horizons indicate a spodic character. Also, the fact that SiBCS criteria classify the studied soils as Espodossolos indicates the relevance of establishing limits for chemical attributes, which would adjust the taxonomy of soils with a spodic character according to their pedogenesi

Attomolar Detection of Botulinum Toxin Type A in Complex Biological Matrices

BACKGROUND: A highly sensitive, rapid and cost efficient method that can detect active botulinum neurotoxin (BoNT) in complex biological samples such as foods or serum is desired in order to 1) counter the potential bioterrorist threat 2) enhance food safety 3) enable future pharmacokinetic studies in medical applications that utilize BoNTs. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a botulinum neurotoxin serotype A assay with a large immuno-sorbent surface area (BoNT/A ALISSA) that captures a low number of toxin molecules and measures their intrinsic metalloprotease activity with a fluorogenic substrate. In direct comparison with the "gold standard" mouse bioassay, the ALISSA is four to five orders of magnitudes more sensitive and considerably faster. Our method reaches attomolar sensitivities in serum, milk, carrot juice, and in the diluent fluid used in the mouse assay. ALISSA has high specificity for the targeted type A toxin when tested against alternative proteases including other BoNT serotypes and trypsin, and it detects the holotoxin as well as the multi-protein complex form of BoNT/A. The assay was optimized for temperature, substrate concentration, size and volume proportions of the immuno-sorbent matrix, enrichment and reaction times. Finally, a kinetic model is presented that is consistent with the observed improvement in sensitivity. CONCLUSIONS/SIGNIFICANCE: The sensitivity, specificity, speed and simplicity of the BoNT ALISSA should make this method attractive for diagnostic, biodefense and pharmacological applications

Caracterização micromorfológica de Espodossolos no Rio de Janeiro.

A pedogênese de solos de ambientes costeiros ainda é pouco estudada. Dentre os solos que ocorrem nesse ambiente podem ser destacados os Espodossolos. A investigação de padrões micromorfológicos de Espodossolos pode contribuir para uma maior compreensão dos processos pedogenéticos que ocorrem nestes solos. Nesse sentido o presente estudo procurou avaliar através da técnica de micromorfologia diferentes formas de matéria orgânica e sua evolução dentro dos processos pedogenéticos existentes em Espodossolos. Para isso foram selecionados dois perfis de Espodossolos no estado do Rio de Janeiro. Os perfis foram descritos e caracterizados segundo (Santos et al., 2013) e alguns horizontes foram selecionados para a realização da descrição micromorfológica. As fotomicrografias identificaram matéria orgânica monomórfica e polimórfica, a primeira associada a revestimentos orgânicos na superfície do esqueleto, e quase revestimentos da porosidade entre grãos. A matéria orgânica polimórfica foi observada nos horizontes que apresentavam resíduos materiais vegetais e descoloramento associados à maior ou menor presença de ferro e matéria orgânica. Para os dois perfis foram identificados processos de eluviação e dissolução demonstrando que a análise micromorfológica é uma ferramenta de grande expressão para processos da matéria orgânica em solos formados neste ambiente

Caracterização e classificação de solos em uma topossequência sobre calcário na Serra da Bodoquena, MS.

A serra da Bodoquena, localizada no Estado do Mato Grosso do Sul, apresenta particularidades nos seus solos, que diferem de outras regiões do bioma cerradopantanal. Este trabalho teve como objetivo ampliar o conhecimento dos solos formados sobre calcário, por meio da caracterização dos seus atributos físicos, químicos, mineralógicos e da matéria orgânica. Foi selecionada uma topossequência sobre calcário, onde foram abertas trincheiras no topo (P1), terço inferior (P2), sopé (P3) e baixada (P4 e P5). Os perfis foram descritos morfologicamente e analisados os atributos físicos, químicos e mineralógicos dos horizontes. De acordo com o Sistema Brasileiro de Classificação de Solos, os solos estudados foram classificados como: (P1) Organossolo Fólico Sáprico lítico - OOs; (P2) Chernossolo Háplico Órtico típico - MXo; (P3) Chernossolo Argilúvico Órtico típico - MTo; (P4) Gleissolo Melânico Carbonático chernossólico - GMk1; e (P5) Gleissolo Melânico Carbonático organossólico - GMk2. Todos os perfis estudados apresentaram cores escuras nos horizontes superficiais e mais avermelhadas ou acinzentadas em profundidade, em razão da drenagem, sempre associados com elevados valores de saturação por bases e tendo o cálcio como cátion predominante no complexo sortivo. Das frações húmicas, a humina representou a maior fração do carbono orgânico em todos os solos. A análise mineralógica constatou a presença de calcita na fração areia nos perfis GMk1 e GMk2 e caulinita, illita e montmorilonita,na fração argila de todos os solos. A ocorrência do Organossolo Fólico em ambiente não altimontano, diferente do relatado pelo Sistema Brasileiro de Classificação de Solos, sugere maior amplitude das condições ambientais para a ocorrência dessa subordem

Biallelic Loss-of-Function Variants in BICD1 Are Associated with Peripheral Neuropathy and Hearing Loss

Hearing loss and peripheral neuropathy are two clinical entities that are genetically and phenotypically heterogeneous and sometimes co-occurring. Using exome sequencing and targeted segregation analysis, we investigated the genetic etiology of peripheral neuropathy and hearing loss in a large Ashkenazi Jewish family. Moreover, we assessed the production of the candidate protein via western blotting of lysates from fibroblasts from an affected individual and an unaffected control. Pathogenic variants in known disease genes associated with hearing loss and peripheral neuropathy were excluded. A homozygous frameshift variant in the BICD1 gene, c.1683dup (p.(Arg562Thrfs*18)), was identified in the proband and segregated with hearing loss and peripheral neuropathy in the family. The BIDC1 RNA analysis from patient fibroblasts showed a modest reduction in gene transcripts compared to the controls. In contrast, protein could not be detected in fibroblasts from a homozygous c.1683dup individual, whereas BICD1 was detected in an unaffected individual. Our findings indicate that bi-allelic loss-of-function variants in BICD1 are associated with hearing loss and peripheral neuropathy. Definitive evidence that bi-allelic loss-of-function variants in BICD1 cause peripheral neuropathy and hearing loss will require the identification of other families and individuals with similar variants with the same phenotype