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Microglial conditioned medium promotes survival and development of cultured mesencephalic neurons from embryonic rat
K Nagata1, N Takei, K Nakajima
1Department of Neurochemistry, National Institute of Neuroscience, Tokyo, Japan.
Abstract:
We previously reported that microglial conditioned medium (Mic-CM) has a neurotrophic effect on cultured rat neocortical neurons [Nakajima et al. (1989): Biomed Res 10:411-423]. In order to investigate the interaction between microglia and neurons in more detail, we determined the effects of Mic-CM on the primary cultured mesencephalic neurons from 16-day embryonic rats. The addition of Mic-CM to the culture medium significantly enhanced the survivability of neurons and promoted neurite extension in a low cell-density culture condition. In a high cell-density culture condition, Mic-CM markedly increased dopamine uptake, which was quantified by assessing the specific [3H]dopamine uptake, and also increased the dopamine content of cultured cells. Furthermore, the number of mesencephalic dopaminergic neurons, which was determined by quantitative analysis of tyrosine hydroxylase (TH)-immunoreactive cells, increased significantly in the presence of Mic-CM. These results suggest that Mic-CM enhances survival or maturation of TH-positive neurons present in cultures of the embryonic mesencephalon and that these neurotrophic effects may be due to a diffusible factor(s) from microglia.
Insights
Microglial conditioned medium (Mic-CM) supports neuron survival and growth. It also boosts dopamine uptake and the number of dopaminergic neurons in developing rat brains.
Area of Science:
- Neuroscience
- Cell Biology
- Developmental Biology
Background:
- Microglia are key immune cells in the central nervous system.
- Microglial conditioned medium (Mic-CM) has previously shown neurotrophic effects on neurons.
Purpose of the Study:
- To investigate the detailed effects of Mic-CM on primary cultured mesencephalic neurons from embryonic rats.
- To understand the interaction between microglia and neurons.
Main Methods:
- Primary mesencephalic neurons from 16-day embryonic rats were cultured.
- The effects of Mic-CM on neuronal survival, neurite extension, dopamine uptake, and dopamine content were assessed.
- Tyrosine hydroxylase (TH)-immunoreactive cells were quantified to determine dopaminergic neuron numbers.
Main Results:
- Mic-CM significantly enhanced neuronal survivability and neurite extension in low-density cultures.
- In high-density cultures, Mic-CM increased specific [3H]dopamine uptake and dopamine content.
- The number of TH-immunoreactive (dopaminergic) neurons significantly increased in the presence of Mic-CM.
Conclusions:
- Mic-CM promotes the survival and maturation of dopaminergic neurons in the embryonic mesencephalon.
- These neurotrophic effects are likely mediated by diffusible factors secreted by microglia.