Dynamic observation of rat hippocampal neuronal death process by hopping probe ion conductance microscopy

doi:10.3969/j.issn.2095-4344.2013.15.001

Abstract

Abstract:

BACKGROUND: The molecular mechanism underlying neuronal cell death has been made some progress in the past years. However, the cell morphology dynamics after neuron death is still missing because of technical shortcomings. Even the time required for the execution of the death program is still unknown. However, neurons cultured in vitro can be observed continually in a non-contact manner by a hopping probe ion conductance microscopy, therefore the morphology changes of a death process can be obtained.
OBJECTIVE: To dynamically observe morphological changes of rat hippocampus neurons during a cell necrotic process by the hopping probe ion conductance microscopy.
METHODS: Primary Wistar rat hippocampal neurons were used for the experiment, and cell necrosis was induced by 1 mmol/L hydrogen peroxide solution for 30 minutes. Neurons without hydrogen peroxide-treatment were used as control. The morphology of neuronal soma and neurites were continually imaged by the hopping probe ion conductance microscopy after hydrogen peroxide treatment, and the height and volume of a neuron were simultaneously measured.
RESULTS AND CONCLUSION: Continuous imaging for 9 hours after hydrogen peroxide treatment under the hopping probe ion conductance microscopy showed that: Hippocampal neurons gradually transformed from a normal triangular or fusiform shape to a more spherical shape due to cell swelling. Axon beading and dendritic beading were gradually formed after scanning, and finally the neurites were ruptured and dissolved. Measurement of the cell height and volume showed that: The height and volume of hydrogen peroxide-treated neurons elevated after 1 hour of scanning, and continued increasing up to 7 hours after scanning (P < 0.05). The final height and volume were approximately twice as beginning. The increase of height and volume manifested as linear correlation with time within 1 to 7 hours by linear-regression analysis (cell height b=0.15, P < 0.005; cell volume b=0.17, P < 0.05). Meanwhile, no significant morphological change was observed during the time of scanning a normal neuron. It is successful to observe necrotic process of hippocampus neurons by the hopping probe ion conductance microscopy. Higher-quality cell image and more accurate information about cell height and volume can be obtained.

Key words: tissue construction, nerve tissue construction, neurons, necrosis, hopping probe ion conductance microscopy, hydrogen peroxide, hippocampus, ultrastructure, rats

CLC Number:

R318

Yang Guo-wei, Zhu Hui, Zhou Zi-wei, Li Ying, Zhang Yan-jun, Zhang Jian-ning. Dynamic observation of rat hippocampal neuronal death process by hopping probe ion conductance microscopy[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(15): 2661-2668.

Figures/Tables 10

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