AFM Imaging Reveals MicroRNA-132 to be a Positive Regulator of Synaptic Functions.

The modification of synaptic and neural connections in adults, including the formation and removal of synapses, depends on activity-dependent synaptic and structural plasticity. MicroRNAs (miRNAs) play crucial roles in regulating these changes by targeting specific genes and regulating their expression. The fact that somatic and dendritic activity in neurons often occurs asynchronously highlights the need for spatial and dynamic regulation of protein synthesis in specific milieu and cellular loci. MicroRNAs, which can show distinct patterns of enrichment, help to establish the localized distribution of plasticity-related proteins. The recent study using atomic force microscopy (AFM)-based nanoscale imaging reveals that the abundance of miRNA(miR)-134 is inversely correlated with the functional activity of dendritic spine structures. However, the miRNAs that are selectively upregulated in potentiated synapses, and which can thereby support prospective changes in synaptic efficacy, remain largely unknown. Using AFM force imaging, significant increases in miR-132 in the dendritic regions abutting functionally-active spines is discovered. This study provides evidence for miR-132 as a novel positive miRNA regulator residing in dendritic shafts, and also suggests that activity-dependent miRNAs localized in distinct sub-compartments of neurons play bi-directional roles in controlling synaptic transmission and synaptic plasticity.

A Pilot Establishment of the Job-Exposure Matrix of Lead Using the Standard Process Code of Nationwide Exposure Databases in Korea.

Background: The purpose of this study is to construct a job-exposure matrix for lead that accounts for industry and work processes within industries using a nationwide exposure database. Methods: We used the work environment measurement data (WEMD) of lead monitored nationwide from 2015 to 2016. Industrial hygienists standardized the work process codes in the database to 37 standard process and extracted key index words for each process. A total of 37 standardized process codes were allocated to each measurement based on an automated key word search based on the degree of agreement between the measurement information and the standard process index. Summary statistics, including the arithmetic mean, geometric mean, and 95th percentile level (X95), was calculated according to industry, process, and industry process. Using statistical parameters of contrast and precision, we compared the similarity of exposure groups by industry, process, and industry process. Results: The exposure intensity of lead was estimated for 583 exposure groups combined with 128 industry and 35 process. The X95 value of the "casting" process of the "manufacture of basic precious and non-ferrous metals" industry was 53.29 µg/m3, exceeding the occupational exposure limit of 50 µg/m3. Regardless of the limitation of the minimum number of samples in the exposure group, higher contrast was observed when the exposure groups were by industry process than by industry or process. Conclusion: We evaluated the exposure intensities of lead by combination of industry and process. The results will be helpful in determining more accurate information regarding exposure in lead-related epidemiological studies.

Quantification of a Neurological Protein in a Single Cell Without Amplification.

Proteins are key biomolecules that not only play various roles in the living body but also are used as biomarkers. If these proteins can be quantified at the level of a single cell, understanding the role of proteins will be deepened and diagnosing diseases and abnormality will be further upgraded. In this study, we quantified a neurological protein in a single cell using atomic force microscopy (AFM). After capturing specifically disrupted-in-schizophrenia 1 (DISC1) in a single cell onto a microspot immobilizing the corresponding antibody on the surface, force mapping with AFM was followed to visualize individual DISC1. Although a large variation of the number of DISC1 in a cell was observed, the average number is 4.38 × 103, and the number agrees with the ensemble-averaged value. The current AFM approach for the quantitative analysis of proteins in a single cell should be useful to study molecular behavior of proteins in depth and to follow physiological change of individual cells in response to external stimuli.

Force Mapping Reveals the Spatial Distribution of Individual Proteins in a Neuron.

Conventional methods for studying the spatial distribution and expression level of proteins within neurons have primarily relied on immunolabeling and/or signal amplification. Here, we present an atomic force microscopy (AFM)-based nanoscale force mapping method, where Anti-LIMK1-tethered AFM probes were used to visualize individual LIMK1 proteins in cultured neurons directly through force measurements. We observed that the number density of LIMK1 decreased in neuronal somas after the cells were depolarized. We also elucidated the spatial distribution of LIMK1 in single spine areas and found that the protein predominantly locates at heads of spines rather than dendritic shafts. The study demonstrates that our method enables unveiling of the abundance and spatial distribution of a protein of interest in neurons without signal amplification or labeling. We expected that this approach should facilitate the studies of protein expression phenomena in depth in a wide range of biological systems.