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N-methyl-D-aspartate receptor activation and Ca2+ account for poor pyramidal cell structure in hippocampal slices
Abstract:
The CA1 pyramidal cells appear damaged in micrographs of guinea pig hippocampal slices incubated in normal physiological buffer at 36-37 degrees C. This is remedied if slices are incubated in modified buffers for the first 45 min. Cell morphology is improved if this buffer is devoid of added Ca2+ and much improved if it contains N-methyl-D-aspartate (NMDA) receptor antagonists or 0 mM Ca2+ and 10 mM Mg2+. The cells then appear similar to CA1 pyramidal cells in situ. These findings support the notion that NMDA receptor activation and Ca2+, acting in the period immediately after slice preparation, permanently damage CA1 pyramidal cells in vitro.
Insights
Newly prepared guinea pig hippocampal slices show CA1 pyramidal cell damage. This damage is prevented by using modified buffers lacking calcium or containing NMDA receptor antagonists during initial incubation.
Area of Science:
- Neuroscience
- Cell Biology
- Physiology
Background:
- CA1 pyramidal cells are crucial for hippocampal function.
- Slice preparation can induce cellular damage in vitro.
- The role of specific ions and receptors in this damage is not fully understood.
Purpose of the Study:
- To investigate the factors contributing to CA1 pyramidal cell damage in hippocampal slices.
- To identify protective buffer conditions for preserving cell morphology post-preparation.
Main Methods:
- Incubation of guinea pig hippocampal slices in various physiological buffers.
- Microscopic examination of CA1 pyramidal cell morphology.
- Testing buffers with modified calcium (Ca2+) and magnesium (Mg2+) concentrations, and N-methyl-D-aspartate (NMDA) receptor antagonists.
Main Results:
- Normal physiological buffer at 36-37°C caused CA1 pyramidal cell damage.
- Modified buffers, especially those lacking Ca2+ or containing NMDA receptor antagonists, significantly improved cell morphology.
- Incubation in 0 mM Ca2+ and 10 mM Mg2+ buffer resulted in cells resembling in situ morphology.
Conclusions:
- NMDA receptor activation and Ca2+ contribute to permanent damage of CA1 pyramidal cells shortly after slice preparation.
- Specific buffer modifications can protect CA1 pyramidal cells during the critical post-preparation period.
- These findings highlight the importance of controlled incubation conditions for in vitro hippocampal slice studies.