Corticofugal projection neurons are key components in connecting the neocortex and the subcortical regions. In the barrel field, these neurons have various projection targets and play crucial roles in the rodent whisker sensorimotor system. However, the projection features of corticofugal projection neurons at the single-axon level are far from comprehensive elucidation. Based on a brain-wide positioning system with high-resolution imaging for Thy1-GFP M-line mice brains, we reconstructed and analyzed more than one hundred corticofugal projection neurons in both layer V and VI of barrel cortex. The dual-color imaging made it possible to locate the neurons’ somata, trace their corresponding dendrites and axons and then distinguish the neurons as L5 type I/II or L6 type. The corticofugal projection pattern showed significant diversity across individual neurons. Usually, the L5 type I neurons have greater multi-region projection potential. The thalamus and the midbrain are the most frequent projection targets among the investigated multidirectional projection neurons, and the hypothalamus is particularly unique in that it only appears in multidirectional projection situations. Statistically, the average branch length of apical dendrites in multi-region projection groups is larger than that of single-region projection groups. This study demonstrated a single-axon-level analysis for barrel corticofugal projection neurons, which could provide a micro-anatomical basis for interpreting whisker sensorimotor circuit function.

Individual cells play essential roles in the biological processes of the brain. The number of neurons changes during both normal development and disease progression. High-resolution imaging has made it possible to directly count cells. However, the automatic and precise segmentation of touching cells continues to be a major challenge for massive and highly complex datasets. Thus, an integrative cut (iCut) algorithm, which combines information regarding spatial location and intervening and concave contours with the established normalized cut, has been developed. iCut involves two key steps: (1) a weighting matrix is first constructed with the abovementioned information regarding the touching cells and (2) a normalized cut algorithm that uses the weighting matrix is implemented to separate the touching cells into isolated cells. This novel algorithm was evaluated using two types of data: the open SIMCEP benchmark dataset and our micro-optical imaging dataset from a Nissl-stained mouse brain. It has achieved a promising recall/precision of 91.2 ± 2.1%/94.1 ± 1.8% and 86.8 ± 4.1%/87.5 ± 5.7%, respectively, for the two datasets. As quantified using the harmonic mean of recall and precision, the accuracy of iCut is higher than that of some state-of-the-art algorithms. The better performance of this fully automated algorithm can benefit studies of brain cytoarchitecture.

The accumulation of amyloid β peptide (Aβ) in the brain is hypothesized to be the major factor driving Alzheimer’s disease (AD) pathogenesis. Mounting evidence suggests that astrocytes are the primary target of Aβ neurotoxicity. Aβ is known to interfere with multiple calcium fluxes, thus disrupting the calcium homeostasis regulation of astrocytes, which are likely to produce calcium oscillations. Ca²⁺ dyshomeostasis has been observed to precede the appearance of clinical symptoms of AD; however, it is experimentally very difficult to investigate the interactions of many mechanisms. Given that Ca²⁺ disruption is ubiquitously involved in AD progression, it is likely that focusing on Ca²⁺ dysregulation may serve as a potential therapeutic approach to preventing or treating AD, while current hypotheses concerning AD have so far failed to yield curable therapies. For this purpose, we derive and investigate a concise mathematical model for Aβ-mediated multi-pathway astrocytic intracellular Ca²⁺ dynamics. This model accounts for how Aβ affects various fluxes contributions through voltage-gated calcium channels, Aβ-formed channels and ryanodine receptors. Bifurcation analysis of Aβ level, which reflected the corresponding progression of the disease, revealed that Aβ significantly induced the increasing [Ca²⁺]_i and frequency of calcium oscillations. The influence of inositol 1,4,5-trisphosphate production (IP₃) is also investigated in the presence of Aβ as well as the impact of changes in resting membrane potential. In turn, the Ca²⁺ flux can be considerably changed by exerting specific interventions, such as ion channel blockers or receptor antagonists. By doing so, a “combination therapy” targeting multiple pathways simultaneously has finally been demonstrated to be more effective. This study helps to better understand the effect of Aβ, and our findings provide new insight into the treatment of AD.

Astrocytic fine processes are the most minor structures of astrocytes but host much of the Ca²⁺ activity. These localized Ca²⁺ signals spatially restricted to microdomains are crucial for information processing and synaptic transmission. However, the mechanistic link between astrocytic nanoscale processes and microdomain Ca²⁺ activity remains hazily understood because of the technical difficulties in accessing this structurally unresolved region. In this study, we used computational models to disentangle the intricate relations of morphology and local Ca²⁺ dynamics involved in astrocytic fine processes. We aimed to answer: 1) how nano-morphology affects local Ca²⁺ activity and synaptic transmission, 2) and how fine processes affect Ca²⁺ activity of large process they connect. To address these issues, we undertook the following two computational modeling: 1) we integrated the in vivo astrocyte morphological data from a recent study performed with super-resolution microscopy that discriminates sub-compartments of various shapes, referred to as nodes and shafts to a classic IP₃R-mediated Ca²⁺ signaling framework describing the intracellular Ca²⁺ dynamics, 2) we proposed a node-based tripartite synapse model linking with astrocytic morphology to predict the effect of structural deficits of astrocytes on synaptic transmission. Extensive simulations provided us with several biological insights: 1) the width of nodes and shafts could strongly influence the spatiotemporal variability of Ca²⁺ signals properties but what indeed determined the Ca²⁺ activity was the width ratio between nodes and shafts, 2) the connectivity of nodes to larger processes markedly shaped the Ca²⁺ signal of the parent process rather than nodes morphology itself, 3) the morphological changes of astrocytic part might potentially induce the abnormality of synaptic transmission by affecting the level of glutamate at tripartite synapses. Taken together, this comprehensive model which integrated theoretical computation and in vivo morphological data highlights the role of the nanomorphology of astrocytes in signal transmission and its possible mechanisms related to pathological conditions.