Synaptic plasticity in human thalamocortical assembloids.

Synaptic plasticities, such as long-term potentiation (LTP) and depression (LTD), tune synaptic efficacy and are essential for learning and memory. Current studies of synaptic plasticity in humans are limited by a lack of adequate human models. Here, we modeled the thalamocortical system by fusing human induced pluripotent stem cell-derived thalamic and cortical organoids. Single-nucleus RNA-sequencing revealed that most cells in mature thalamic organoids were glutamatergic neurons. When fused to form thalamocortical assembloids, thalamic and cortical organoids formed reciprocal long-range axonal projections and reciprocal synapses detectable by light and electron microscopy, respectively. Using whole-cell patch-clamp electrophysiology and two-photon imaging, we characterized glutamatergic synaptic transmission. Thalamocortical and corticothalamic synapses displayed short-term plasticity analogous to that in animal models. LTP and LTD were reliably induced at both synapses; however, their mechanisms differed from those previously described in rodents. Thus, thalamocortical assembloids provide a model system for exploring synaptic plasticity in human circuits.

Jedi-1/MEGF12-mediated phagocytosis controls the pro-neurogenic properties of microglia in the ventricular-subventricular zone.

Microglia are the primary phagocytes in the central nervous system and clear dead cells generated during development or disease. The phagocytic process shapes the microglia phenotype, which affects the local environment. A unique population of microglia resides in the ventricular-subventricular zone (V-SVZ) of neonatal mice, but how they influence the neurogenic niche is not well understood. Here, we demonstrate that phagocytosis contributes to a pro-neurogenic microglial phenotype in the V-SVZ and that these microglia phagocytose apoptotic cells via the engulfment receptor Jedi-1. Deletion of Jedi-1 decreases apoptotic cell clearance, triggering a neuroinflammatory microglia phenotype that resembles dysfunctional microglia in neurodegeneration and aging and that reduces neural precursor proliferation via elevated interleukin-1ß signaling; interleukin-1 receptor inhibition rescues precursor proliferation in vivo. Together, these results reveal a critical role for Jedi-1 in connecting microglial phagocytic activity to the maintenance of a pro-neurogenic phenotype in the developing V-SVZ.

Jedi-1/MEGF12-mediated phagocytosis controls the pro-neurogenic properties of microglia in the ventricular-subventricular zone.

Microglia are the primary phagocytes in the central nervous system and are responsible for clearing dead cells generated during development or disease. The phagocytic process shapes the phenotype of the microglia, which affects the local environment. A unique population of microglia reside in the ventricular-subventricular zone (V-SVZ) of neonatal mice, but how they influence this neurogenic niche is not well-understood. Here, we demonstrate that phagocytosis creates a pro-neurogenic microglial phenotype in the V-SVZ and that these microglia phagocytose apoptotic cells via the engulfment receptor Jedi-1. Deletion of Jedi-1 decreases apoptotic cell clearance, triggering the development of a neuroinflammatory phenotype, reminiscent of neurodegenerative and-age-associated microglia, that reduces neural precursor proliferation via elevated interleukin (IL)-1ß signaling; inhibition of IL-1 receptor rescues precursor proliferation in vivo. Together, these results reveal a critical role for Jedi-1 in connecting microglial phagocytic activity to a phenotype that promotes neurogenesis in the developing V-SVZ.

Jedi-1 deficiency increases sensory neuron excitability through a non-cell autonomous mechanism.

The dorsal root ganglia (DRG) house the primary afferent neurons responsible for somatosensation, including pain. We previously identified Jedi-1 (PEAR1/MEGF12) as a phagocytic receptor expressed by satellite glia in the DRG involved in clearing apoptotic neurons during development. Here, we further investigated the function of this receptor in vivo using Jedi-1 null mice. In addition to satellite glia, we found Jedi-1 expression in perineurial glia and endothelial cells, but not in sensory neurons. We did not detect any morphological or functional changes in the glial cells or vasculature of Jedi-1 knockout mice. Surprisingly, we did observe changes in DRG neuron activity. In neurons from Jedi-1 knockout (KO) mice, there was an increase in the fraction of capsaicin-sensitive cells relative to wild type (WT) controls. Patch-clamp electrophysiology revealed an increase in excitability, with a shift from phasic to tonic action potential firing patterns in KO neurons. We also found alterations in the properties of voltage-gated sodium channel currents in Jedi-1 null neurons. These results provide new insight into the expression pattern of Jedi-1 in the peripheral nervous system and indicate that loss of Jedi-1 alters DRG neuron activity indirectly through an intercellular interaction between non-neuronal cells and sensory neurons.

Peripheral myelin protein 22 modulates store-operated calcium channel activity, providing insights into Charcot-Marie-Tooth disease etiology.

Charcot-Marie-Tooth (CMT) disease is a peripheral neuropathy associated with gene duplication and point mutations in the peripheral myelin protein 22 (PMP22) gene. However, the role of PMP22 in Schwann cell physiology and the mechanisms by which PMP22 mutations cause CMT are not well-understood. On the basis of homology between PMP22 and proteins associated with modulation of ion channels, we hypothesized that PMP22 alters ion channel activity. Using whole-cell electrophysiology, we show here that heterologous PMP22 expression increases the amplitude of currents similar to those ascribed to store-operated calcium (SOC) channels, particularly those involving transient receptor canonical channel 1 (TrpC1). These channels help replenish Ca2+ in the endoplasmic reticulum (ER) following stimulus-induced depletion. Currents with similar properties were recorded in WT but not pmp22-/- mouse Schwann cells. Heterologous expression of the CMT-associated PMP22_L16P variant, which fails to reach the plasma membrane and localizes to the ER, led to larger currents than WT PMP22. Similarly, Schwann cells isolated from Trembler J (TrJ; PMP22_L16P) mice had larger currents than WT littermates. Calcium imaging in live nerves and cultured Schwann cells revealed elevated intracellular Ca2+ in TrJ mice compared with WT. Moreover, we found that PMP22 co-immunoprecipitated with stromal interaction molecule 1 (STIM1), the Ca2+ sensor SOC channel subunit in the ER. These results suggest that in the ER, PMP22 interacts with STIM1 and increases Ca2+ influx through SOC channels. Excess or mutant PMP22 in the ER may elevate intracellular Ca2+ levels, which could contribute to CMT pathology.

Synthesis and characterization of non-hydrolysable diphosphoinositol polyphosphate second messengers.

The diphosphoinositol polyphosphates (PP-IPs) are a central group of eukaryotic second messengers. They regulate numerous processes, including cellular energy homeostasis and adaptation to environmental stresses. To date, most of the molecular details in PP-IP signalling have remained elusive, due to a lack of appropriate methods and reagents. Here we describe the expedient synthesis of methylene-bisphosphonate PP-IP analogues. Their characterization revealed that the analogues exhibit significant stability and mimic their natural counterparts very well. This was further confirmed in two independent biochemical assays, in which our analogues potently inhibited phosphorylation of the protein kinase Akt and hydrolytic activity of the Ddp1 phosphohydrolase. The non-hydrolysable PP-IPs thus emerge as important tools and hold great promise for a variety of applications.

[Behavior of immatures Aedes aegypti in the coast State of São Paulo, Brazil]. / Comportamento de formas imaturas de Aedes aegypti, no litoral do Estado de São Paulo.

INTRODUCTION: In a region of high dengue incidence, on the coast of the State of São Paulo, 9 areas were selected to evaluate the behavior of immature Aedes aegypti. METHODS: The 9 areas were grouped into 4 strata according to soil use and occupation. Larvae and pupas were collected in a sample of approximately 500 buildings in each area. RESULTS: Although tires and canvas presented the highest positive rates for Aedes aegypti, drains and other containers fixed to the buildings were highly predominant among positive containers; 32 to 76% of the positive containers in the 4 study strata. Public areas of apartment buildings and large non-residential premises presented the highest positive rates for Aedes Aegypti, while apartments presented the lowest. Infestation levels were greater in residential areas with predominance of apartment buildings, where 76% of the breeding sites detected were containers fixed to the buildings. CONCLUSIONS: This knowledge is an important tool in the control strategy, since it reinforces the need for special attention regarding certain types of buildings and the adjustment of technical norms for pluvial water drains and improvement of building maintenance. Moreover, systematic observations are required to follow-up the occupancy dynamic of different buildings and containers by Aedes aegypti and the incorporation of this knowledge in the control of vectors in the region.

Comportamento de formas imaturas de Aedes aegypti, no litoral do Estado de São Paulo / Behavior of immatures Aedes aegypti in the coast State of São Paulo, Brazil

INTRODUÇÃO: Em região de alta incidência de dengue, no litoral do Estado de São Paulo, selecionaram-se 9 áreas, com objetivo de avaliar o comportamento de formas imaturas de Aedes aegypti. MÉTODOS: As 9 áreas foram agrupadas em 4 estratos, diferenciados pelo uso e ocupação do solo. Foram coletadas larvas e pupas numa amostra de cerca de 500 imóveis em cada área. RESULTADOS: Apesar do pneu e lona apresentarem as maiores taxas de positividade para Aedes aegypti, o ralo, juntamente com outros recipientes fixos nas edificações foram altamente predominantes entre os recipientes positivos (32 a 76 por cento dos recipientes positivos). As áreas coletivas de prédios e os imóveis não residenciais de grande porte apresentaram as maiores taxas de positividade para Aedes aegypti enquanto os apartamentos, as menores. Os níveis de infestação foram maiores na área residencial com predominância de prédios de apartamentos, onde 76 por cento dos criadouros detectados foram recipientes fixos nas edificações. CONCLUSÕES: Esses conhecimentos são importantes subsídios para a estratégia de controle, pois reforçam a necessidade de atenção especial para determinados tipos de imóveis, bem como da adequação da norma técnica de ralo de água pluvial e da melhoria de manutenção das edificações. Além disso, são necessárias observações sistemáticas que permitam acompanhar a dinâmica de ocupação de diferentes imóveis e recipientes por Aedes aegypti e a incorporação desses conhecimentos nas ações de controle do vetor na região.

INTRODUCTION: In a region of high dengue incidence, on the coast of the State of São Paulo, 9 areas were selected to evaluate the behavior of immature Aedes aegypti. METHODS: The 9 areas were grouped into 4 strata according to soil use and occupation. Larvae and pupas were collected in a sample of approximately 500 buildings in each area. RESULTS: Although tires and canvas presented the highest positive rates for Aedes aegypti, drains and other containers fixed to the buildings were highly predominant among positive containers; 32 to 76 percent of the positive containers in the 4 study strata. Public areas of apartment buildings and large non-residential premises presented the highest positive rates for Aedes Aegypti, while apartments presented the lowest. Infestation levels were greater in residential areas with predominance of apartment buildings, where 76 percent of the breeding sites detected were containers fixed to the buildings. CONCLUSIONS: This knowledge is an important tool in the control strategy, since it reinforces the need for special attention regarding certain types of buildings and the adjustment of technical norms for pluvial water drains and improvement of building maintenance. Moreover, systematic observations are required to follow-up the occupancy dynamic of different buildings and containers by Aedes aegypti and the incorporation of this knowledge in the control of vectors in the region.

Persistência da transmissão de Schistosoma mansoni em localidade de Itariri, na região do Vale do Ribeira, São Paulo, Brasil / Persistent transmission of Schistosoma mansoni Itariri location in the Vale do Ribeira, Sao Paulo, Brazil

Introdução A transmissão de esquistossomose é considerada um problema de saúde pública em várias regiões do Brasil, entre elas, a região do Vale do Ribeira, Estado de São Paulo. Em 2009, um censo coprológico com alunos de 6 a 14 anos na escola estadual do bairro de Ana Dias, no município de Itariri, detectou 34 casos de esquistossomose (16,8 por cento de positividade) e sinalizou a existência de transmissão autóctone importante naquela área. Objetivo Aprofundar os conhecimentos sobre a transmissão de esquistossomose no bairro de Ana Dias. Métodos Aplicou-se um formulário aos pacientes, a fim de conhecer seus hábitos em relação às coleções hídricas e condições de saneamento das suas residências. As coleções hídricas do bairro são pesquisadas rotineiramente e os planorbídeos encontrados, examinados para verificar a existência de cercárias. Dos dados obtidos foi realizado um levantamento de coleções com presença de planorbídeos infectados por S. mansoni no período de 2000 a 2010, que foram visitadas e tiveram suas coordenadas anotadas e transferidas para mapas digitalizados. Essas informações foram relacionadas com as obtidas através dos formulários para se conhecer alguns aspectos da transmissão local. Resultados Foi aplicado formulário em 31 pessoas (91,2 por cento do total) e 100 por cento delas declararam ter contato com águas de pelo menos um dos dois principais rios do bairro. Quanto ao saneamento, 50,1 por cento têm o despejo dos dejetos em fossas e valas a céu aberto e 22,6 por cento despejam diretamente nos rios. No período, cerca de 50 por cento das coleções hídricas apresentaram planorbídeos positivos para S. mansoni. Verificou-se que a maioria das coleções positivas são valas de drenagem existentes em bananais e tem ligação direta com os dois rios freqüentados pela população. Nos bananais há casas cujo esgoto é despejado diretamente em valas de drenagem. Conclusões O estudo mostrou que a transmissão autóctone persiste no bairro e ocorre principalmente em função dos rios freqüentados como principal opção de lazer da população. A contaminação dos rios acontece pelos dejetos lançados neles por deficiência do saneamento básico local. Além do trabalho de controle atual, é necessária a realização de atividades regulares que possibilitem a identificação e tratamento dos portadores de esquistossomose, evitando-se a contaminação das coleções.