Upper brainstem cholinergic neurons project to ascending and descending circuits.

BACKGROUND: Based on their anatomical location, rostral projections of nuclei are classified as ascending circuits, while caudal projections are classified as descending circuits. Upper brainstem neurons participate in complex information processing and specific sub-populations preferentially project to participating ascending or descending circuits. Cholinergic neurons in the upper brainstem have extensive collateralizations in both ascending and descending circuits; however, their single-cell projection patterns remain unclear because of the lack of comprehensive characterization of individual neurons. RESULTS: By combining fluorescent micro-optical sectional tomography with sparse labeling, we acquired a high-resolution whole-brain dataset of pontine-tegmental cholinergic neurons (PTCNs) and reconstructed their detailed morphology using semi-automatic reconstruction methods. As the main source of acetylcholine in some subcortical areas, individual PTCNs had abundant axons with lengths up to 60 cm and 5000 terminals and innervated multiple brain regions from the spinal cord to the cortex in both hemispheres. Based on various collaterals in the ascending and descending circuits, individual PTCNs were grouped into four subtypes. The morphology of cholinergic neurons in the pedunculopontine nucleus was more divergent, whereas the laterodorsal tegmental nucleus neurons contained richer axonal branches and dendrites. In the ascending circuits, individual PTCNs innervated the thalamus in three different patterns and projected to the cortex via two separate pathways. Moreover, PTCNs targeting the ventral tegmental area and substantia nigra had abundant collaterals in the pontine reticular nuclei, and these two circuits contributed oppositely to locomotion. CONCLUSIONS: Our results suggest that individual PTCNs have abundant axons, and most project to various collaterals in the ascending and descending circuits simultaneously. They target regions with multiple patterns, such as the thalamus and cortex. These results provide a detailed organizational characterization of cholinergic neurons to understand the connexional logic of the upper brainstem.

The Mesoscopic Connectome of the Cholinergic Pontomesencephalic Tegmentum.

The pontomesencephalic tegmentum, comprising the pedunculopontine nucleus and laterodorsal tegmental nucleus, is involved in various functions via complex connections; however, the organizational structure of these circuits in the whole brain is not entirely clear. Here, combining viral tracing with fluorescent micro-optical sectional tomography, we comprehensively investigated the input and output circuits of two cholinergic subregions in a continuous whole-brain dataset. We found that these nuclei receive abundant input with similar spatial distributions but with different quantitative measures and acquire similar neuromodulatory afferents from the ascending reticular activation system. Meanwhile, these cholinergic nuclei project to similar targeting areas throughout multiple brain regions and have different spatial preferences in 3D. Moreover, some cholinergic connections are unidirectional, including projections from the pedunculopontine nucleus and laterodorsal tegmental nucleus to the ventral posterior complex of the thalamus, and have different impacts on locomotion and anxiety. These results reveal the integrated cholinergic connectome of the midbrain, thus improving the present understanding of the organizational structure of the pontine-tegmental cholinergic system from its anatomical structure to its functional modulation.

Mechanisms of niche-neutrality balancing can drive the assembling of microbial community.

One hotspot of present community ecology is to uncover the mechanisms of community succession. In this study, two popular concepts, niche-neutrality dynamic balancing and co-occurrence network analysis, were integrated to investigate the dispersal dynamics of microbial communities in a freshwater river continuum in subtropical China. Results showed that when habitat conditions were mild and appropriate, such as in the clean upstream river, free of heavy pollution or long-lasting extreme disturbances, stochastic processes could increase species diversities, and organize communities into relatively loosely linked and stable networks with higher modularity and more modules. However, when conditions became degraded under heavy pollution, the influence of neutrality diminished, and niche-based selection imposed more constraints on communities and guided the assembling processes in certain directions: depleting species richness, strengthening interspecies connections and breaking boundaries of modules. Consequently, communities became more sensitive to fluctuations so as to deal with the harsh conditions efficiently. Another interesting finding was that, both as keystone taxa of communities, module hubs were mostly neutrally distributed generalists with high abundances, and were beneficial to many related operational taxonomic units. In contrast, connectors were less abundant and their distributions were more subjected to the environments. Therefore, connectors were probably responsible for the information transmission between microbial communities and environments, as well as between different modules, and thus could restrict the dispersal of microbes and guide the direction of community assembly.

Long-range inputome of cortical neurons containing corticotropin-releasing hormone.

Dissection of the neural circuits of the cerebral cortex is essential for studying mechanisms underlying brain function. Herein, combining a retrograde rabies tracing system with fluorescent micro-optical sectional tomography, we investigated long-range input neurons of corticotropin-releasing hormone containing neurons in the six main cortical areas, including the prefrontal, somatosensory, motor, auditory, and visual cortices. The whole brain distribution of input neurons showed similar patterns to input neurons distributed mainly in the adjacent cortical areas, thalamus, and basal forebrain. Reconstruction of continuous three-dimensional datasets showed the anterior and middle thalamus projected mainly to the rostral cortex whereas the posterior and lateral projected to the caudal cortex. In the basal forebrain, immunohistochemical staining showed these cortical areas received afferent information from cholinergic neurons in the substantia innominata and lateral globus pallidus, whereas cholinergic neurons in the diagonal band nucleus projected strongly to the prefrontal and visual cortex. Additionally, dense neurons in the zona incerta and ventral hippocampus were found to project to the prefrontal cortex. These results showed general patterns of cortical input circuits and unique connection patterns of each individual area, allowing for valuable comparisons among the organisation of different cortical areas and new insight into cortical functions.

Effects of environmental variables on abundance of ammonia-oxidizing communities in sediments of Luotian River, China.

Ammonia-oxidizing communities play important functional roles in the nitrification. However, environmental stresses can significantly affect this process by controlling the abundant communities of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) communities. In this study, we examined the abundance variations of ammonia-oxidizing communities using quantitative polymerase chain reaction (qPCR) and terminal-restriction fragment length polymorphism (T-RFLP) in a typical subtropical river, Luotian County, South Dabie Mountains, China. Clone libraries were conducted to evaluate the community structure and abundance of AOA and AOB in sediments. Results showed that Nitrososphaera sp and Nitrosopumilus sp were the most dominant AOA. The abundance of the AOA and AOB amoA gene ranged from 5.28 × 108 gene copies (g-soil-1) to 2.23 × 108 gene copies (g-soil-1) and 5.45 × 108 gene copies (g-soil-1) to 3.30 × 107 gene copies (g-soil-1), respectively. Five environmental variables, namely, ORP, DO, NO 3 - , Temp, and NH 4 + were played a major function in microbial communities of AOA and AOB in sediments. The T-RFLP profiles of AOA showed that 488 and 116 bp T-RFs were dominated. Overall, the results of this study showed that anthropogenic activities andenvironmental stress in rivers can alter the structure and function of microbes in their variable environment.

A DNA Origami Mechanical Device for the Regulation of Microcosmic Structural Rigidity.

DNA origami makes it feasible to fabricate a tremendous number of DNA nanostructures with various geometries, dimensions, and functionalities. Moreover, an increasing amount of research on DNA nanostructures is focused on biological and biomedical applications. Here, the reversible regulation of microcosmic structural rigidity is accomplished using a DNA origami device in vitro. The designed DNA origami monomer is composed of an internal central axis and an external sliding tube. Due to the external tube sliding, the device transforms between flexible and rigid states. By transporting the device into the liposome, the conformational change of the origami device induces a structural change in the liposome. The results obtained demonstrate that the programmed DNA origami device can be applied to regulate the microcosmic structural rigidity of liposomes. Because microcosmic structural rigidity is important to cell proliferation and function, the results obtained potentially provide a foundation for the regulation of cell microcosmic structural rigidity using DNA nanostructures.