Adaptations to the neuronal culture for researchers at undergraduate institutions

Abstract

Background

The use of rat hippocampal neurons in culture has become an essential tool in neuroscience, enabling detailed study of excitatory synapse organization, neurotransmitter release, and mechanisms of synaptic plasticity. While these cultures provide valuable insights, the physiological relevance of this simplified in vitro system remains an ongoing discussion. Research indicates that cultured hippocampal neurons undergo key maturation processes, including the development of mature dendritic spines, within weeks, mirroring aspects of in vivo development. Importantly, cultured neurons offer unique experimental flexibility, facilitating single-neuron manipulations that is technically challenging or impractical in intact brain slices or with viral vectors. Despite these advantages, establishing cultures with minimal glial support—critical for experiments involving sparse labeling of extracellular proteins for single-particle tracking—often demands substantial time, expertise, and resources, making it difficult to implement in smaller laboratories with limited personnel and funding.

New method

In this study, we present modifications to the standard hippocampal culture protocol designed to improve accessibility and usability in resource-limited settings, such as undergraduate-focused institutions.

Results/Comparison

Our protocol reduces costs, simplifies the culturing process, and minimizes time requirements, supporting robust neuronal cultures with physiological properties comparable to those of traditional methods. These adaptations enable the execution of sophisticated experiments, including single-molecule tracking, in personnel-limited research environments.

Conclusions

This approach highlights the potential for undergraduate institutions to make significant contributions to scientific advancements, rather than being viewed solely as centers for undergraduate training.