α-Enolase binds to human plasminogen on the surface of Bacillus anthracis

α-Enolase of Bacillus anthracis has recently been classified as an immunodominant antigen and a potent virulence factor determinant. α-enolase (2-phospho-D-glycerate hydrolase (EC 4.2.1.11), a key glycolytic metalloenzyme catalyzes the dehydration of D-(+)-2-phosphoglyceric acid to phosphoenolpyruvate. Interaction of surface bound a-enolase with plasminogen has been incriminated in tissue invasion for pathogenesis. B. anthracis α-enolase was expressed in Escherichia coli and the recombinant enzyme was purified to homogeneity that exhibited a Km of 3.3 mM for phosphoenolpyruvate and a Vmax of 0.506 μMmin−1 mg−1. B. anthracis whole cells and membrane vesicles probed with anti-enolase antibodies confirmed the surface localization of α-enolase. The specific interaction of α-enolase with human plasminogen (but not plasmin) evident from ELISA and the retardation in the native gel reinforced its role in plasminogen binding. Putative plasminogen receptors in B. anthracis other than enolase were also observed. This binding was found to be carboxypeptidase sensitive implicating the role of C-terminal lysine residues. The recombinant enolase displayed in vitro laminin binding, an important mammalian extracellular matrix protein. Plasminogen interaction conferred B. anthracis with a potential to in vitro degrade fibronectin and exhibit fibrinolytic phenotype. Therefore, by virtue of its interaction to host plasminogen and extracellular matrix proteins, α-enolase may contribute in augmenting the invasive potential of B. anthracis

Molecular, spatial, and functional single-cell profiling of the hypothalamic preoptic region.

The hypothalamus controls essential social behaviors and homeostatic functions. However, the cellular architecture of hypothalamic nuclei-including the molecular identity, spatial organization, and function of distinct cell types-is poorly understood. Here, we developed an imaging-based in situ cell-type identification and mapping method and combined it with single-cell RNA-sequencing to create a molecularly annotated and spatially resolved cell atlas of the mouse hypothalamic preoptic region. We profiled ~1 million cells, identified ~70 neuronal populations characterized by distinct neuromodulatory signatures and spatial organizations, and defined specific neuronal populations activated during social behaviors in male and female mice, providing a high-resolution framework for mechanistic investigation of behavior circuits. The approach described opens a new avenue for the construction of cell atlases in diverse tissues and organisms

Molecular, spatial, and functional single-cell profiling of the hypothalamic preoptic region

The hypothalamus controls essential social behaviors and homeostatic functions. However, the cellular architecture of hypothalamic nuclei-including the molecular identity, spatial organization, and function of distinct cell types-is poorly understood. Here, we developed an imaging-based in situ cell-type identification and mapping method and combined it with single-cell RNA-sequencing to create a molecularly annotated and spatially resolved cell atlas of the mouse hypothalamic preoptic region. We profiled ~1 million cells, identified ~70 neuronal populations characterized by distinct neuromodulatory signatures and spatial organizations, and defined specific neuronal populations activated during social behaviors in male and female mice, providing a high-resolution framework for mechanistic investigation of behavior circuits. The approach described opens a new avenue for the construction of cell atlases in diverse tissues and organisms

Properties of cells measured with MERFISH

Properties of all cells measured with MERFISH provided as a csv file. “Cell ID” is a unique ID associated with each cell. “Animal ID” is a unique ID associated with animal. “Animal sex” is the gender of the animal in which the cell was imaged. “Behavior” describes the behavioral treatment of the animal. ‘Naïve’ indicates that no treatment was performed. “Bregma” indicates the approximate location of the slice in bregma coordinates. “Centroid X” is the x coordinate of the centroid position for the cell in µm. “Centroid Y” is the y coordinate of the centroid position for the cell in µm. “Cell class” lists the major cell class to which a cell was assigned. A value of 'Ambiguous' represents cells that were identified as putative doublets and were not further analyzed. “Neuron cluster_ID” represents the neuronal cluster to which a cell was assigned. This field is empty if the cell was not a neuron. Columns with a gene name, e.g. ‘Ace2’, contain the expression values for that gene in that cell. Expression values for the 135 genes measured in the combinatorial smFISH run were determined as the total counts per cell divided by the cell volume and scaled by 1000. Expression values for the 21 genes (including Fos) measured in the non-combinatorial, sequential FISH rounds were arbitrary fluorescence units per µm^3, but the same scale is used for all cells. A value of 'NaN' for Fos indicates that this gene was not measured in this animal. These 21 genes are the final genes in the listed genes. Genes named 'Blank-' represent the measurement of barcodes not assigned to any RNA and which serve as blank controls. There are five blank controls

Data from: Molecular, spatial and functional single-cell profiling of the hypothalamic preoptic region

The hypothalamus controls essential social behaviors and homeostatic functions. However, the cellular architecture of hypothalamic nuclei, including the molecular identity, spatial organization, and function of distinct cell types, is poorly understood. Here, we developed an imaging-based cell type identification and mapping method and combined it with single-cell RNA-sequencing to create a molecularly annotated and spatially resolved cell atlas of the mouse hypothalamic preoptic region. We profiled ~1 million cells, identified ~70 neuronal populations characterized by distinct neuromodulatory signatures and spatial organizations, and defined specific neuronal populations activated during key social behaviors in male and female mice, providing a high-resolution framework for mechanistic investigation of behavior circuits. The approach described here opens a new avenue for the construction of cell atlases in diverse tissues and organisms