Cholinergic system is very important for many higher brain functions, including learning and memory. Cholinergic neurons, especially those in the basal forebrain, are specifically susceptible in some neurodegenerative diseases, such as in Alzheimer’s disease (AD). Here, we studied the cholinergic system lesion effects of five familial AD mutations in 5XFAD mice, a transgenic mouse model of AD. Although the cholinergic system has been studied in this mouse model, the cholinergic deficits in AD mice have never been systematically mapped in a whole-brain three-dimensional (3D) reconstruction. Using the 3D reconstruction technology combined with immunohistochemistry (3D-IHC) and design-based stereology, we comprehensively compared the differences of the cholinergic neurons and fibers between the 5XFAD mice and C57BL/6 control mice at different age. Here, we found that the lesion of cholinergic fibers occurred earlier than the cholinergic neuron loss in 5XFAD mice. The cholinergic fiber lesions in the AD mice started sequentially in amygdala, cortex, hippocampus, and then basal forebrain. However, the basal forebrain was the first brain region observed with cholinergic neuron loss at the age of 9 months in 5XFAD mice, whereas such phenomenon first occurred at the age of 15 months in C57BL/6 control mice. Moreover, using 3D reconstruction to compare the lesion of cholinergic system of aged 5XFAD and C57BL/6 control mice, it is intuitive to notice the pathologic regions and severity of lesion. Therefore, the 3D-IHC provides detailed overview of the cholinergic neurons in the whole mouse brain, which will contribute to the study of the developing and pathologic mouse brain.
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We gratefully acknowledge Dr. Qing Tian, Dr. Linqiang Zhu, and Dr. Juan Chen for their helpful discussion and suggestions; Jiaming Liang for the image alignment code; and Shaohua Zhang for the assistance with equipment maintenance. This study was supported by the National Nature Science Foundation of China (Grant Nos. 81130079, 91232302, 81571078, and 3160047).
The authors declare that they have no competing interests.
The original version of this article was revised: The presentation of the name K. A. Henok needs to be updated to “Henok K.A.”. Also in authors contribution, the last sentence should be read as “H.Y., L.P., Y.H., H.K., and Y.L. wrote the manuscript.”
Electronic Supplementary Material
The 3D movie of cholinergic neurons in the mouse brain. Firstly, there are all the ChAT-positive neurons in the hemisphere mouse brain. Then, these neurons in the different brain regions are labeled by different color balls. Last, all the cholinergic fiber lesions in the various brain regions are labeled by different color balls. (MOV 6910 kb).
The code, which was written based on the software of MATLAB R2015b, was used for the alignment of pictures of serial brain sections. (M 1 kb).
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Yan, H., Pang, P., Chen, W. et al. The Lesion Analysis of Cholinergic Neurons in 5XFAD Mouse Model in the Three-Dimensional Level of Whole Brain. Mol Neurobiol 55, 4115–4125 (2018). https://doi.org/10.1007/s12035-017-0621-4
- Alzheimer’s disease
- 5XFAD mouse model
- Cholinergic neurons
- Three dimension
- Whole brain