A PDMS–Agar Hybrid Microfluidic Device for the Investigation of Chemical–Mechanical Associative Learning Behavior of C. elegans

Associative learning is a critical survival trait that promotes behavioral plasticity in response to changing environments. Chemosensation and mechanosensation are important sensory modalities that enable animals to gather information about their internal state and external environment. However, there is a limited amount of research on these two modalities. In this paper, a novel PDMS–agar hybrid microfluidic device is proposed for training and analyzing chemical–mechanical associative learning behavior in the nematode Caenorhabditis elegans. The microfluidic device consisted of a bottom agar gel layer and an upper PDMS layer. A chemical concentration gradient was generated on the agar gel layer, and the PDMS layer served to mimic mechanical stimuli. Based on this platform, C. elegans can perform chemical–mechanical associative learning behavior after training. Our findings indicated that the aversive component of training is the primary driver of the observed associative learning behavior. In addition, the results indicated that the neurotransmitter octopamine is involved in regulating this associative learning behavior via the SER-6 receptor. Thus, the microfluidic device provides a highly efficient platform for studying the associative learning behavior of C. elegans, and it may be applied in mutant screening and drug testing.