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Selective expression of factors preventing cholinergic dedifferentiation
F G DeMello1, M C DeMello, R Hudson
1Department of Neurobiology and Physiology, Northwestern University Institute for Neuroscience, Evanston, IL 60208.
Journal of Neurochemistry
|March 1, 1990
Summary
Chicken retina neurons cultured in vitro initially develop strong cholinergic properties. However, choline acetyltransferase activity declines over time, but this can be prevented using aggregate cultures or conditioned medium.
Area of Science:
- Neuroscience
- Developmental Biology
- Cell Culture
Background:
- Chicken retina neurons in vitro exhibit transient cholinergic properties.
- Maintaining cholinergic function in cultured neurons is crucial for studying neuronal development and function.
Purpose of the Study:
- To investigate the development and maintenance of cholinergic properties in cultured chicken retina neurons.
- To identify factors and conditions that can stabilize cholinergic differentiation in vitro.
Main Methods:
- Culturing chicken retina neurons in stationary (monolayer) and rotary (aggregate) systems.
- Measuring high-affinity choline uptake, [3H]-acetylcholine synthesis and release, and choline acetyltransferase (ChAT) activity.
- Analyzing the effects of conditioned medium from aggregate cultures on monolayer cultures.
Main Results:
- Newly cultured neurons showed high [3H]choline uptake, [3H]-acetylcholine synthesis, and release, with ChAT activity comparable to intact retina.
- Cholinergic function, particularly [3H]ACh release and ChAT activity, declined significantly in stationary cultures over time.
- Maintaining neurons in aggregate cultures or using conditioned medium from these cultures prevented the decline in ChAT activity and stabilized cholinergic differentiation.
Conclusions:
- Cholinergic differentiation in cultured chicken retina neurons is not permanently maintained in standard monolayer cultures.
- The loss of cholinergic properties is linked to a decrease in ChAT activity, not necessarily neuronal loss.
- Aggregate culture conditions or specific factors in conditioned medium can preserve and enhance cholinergic differentiation, mimicking in situ developmental patterns.