期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 158, 期 2, 页码 251-259出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2006.06.006
关键词
brain injury; glycerol-based modified artificial cerebrospinal fluid; electrophysiology; neuronal protection; spinal cord injury
资金
- NCCIH NIH HHS [AT001182] Funding Source: Medline
- NIAAA NIH HHS [AA-11989] Funding Source: Medline
Viable neurons in brain slices are crucial for electrophysiological studies. The present study describes a new method for obtaining viable cells in several regions of the central nervous system including the ventral tegmental area, the hypothalamus, the periaqueductal grey matter and the spinal cord. The essence of the method was to use a modified artificial cerebrospinal fluid (ACSF) in which all NaCl was replaced initially by equi-osmotic glycerol. This modified glycerol-based ACSF was used during slice preparation. The underlying principle for the modification is to prevent the possible acute neurotoxic effects of passive chloride entry, subsequent cell swelling and lysis. This method significantly increased the live/dead ratio in morphology compared to the normal ACSF or sucrose-base ACSF, in which NaCl was replaced by sucrose. An examination of some electrophysiological and pharmacological properties of the neurons in these preparations, by means of current-clamp and voltage-clamp recordings, revealed similar properties of those neurons obtained with the traditional ACSF method. Due to the increase in the number of viable neurons, the new ACSF increases the productivity of experiments. Based on our data, we propose that this glycerol-based solution may protect CNS neurons. (c) 2006 Elsevier B.V. All fights reserved.
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