DNA-mediated assembly of gustatory and neuronal cell and taste signal transmission

Abstract

Cells are surrounded by other cells and interact with each other through close connection. This cell-cell adhesive interaction induces signaling pathways and affects the behavior of cells. Especially, in taste buds, gustatory cells and neuronal cells are closely located and information about taste is transmitted from tongue to brain through neuronal cells. In order to make gustatory and neuronal cells interact like taste buds, it is essential to bring both cells close together. In this study, I investigate DNA-PEG-DPPE polymers as a linker to closely connect gustatory cells and neuronal cells for mimicking actual taste buds with an aim to understand how gustatory and neuronal cells communicate with each other as a response to taste stimuli in taste buds. DNA-PEG-DPPE consists of 1,2-dipalmitoyl-glycerol-3-phosphatidylethanolamine, two thiol modified single stranded oligonucleotides: $(ACTG)_5$-SH and $(CAGT)_5$-SH, and succinimidyl-[(N-maleimidopropionamido)-tetracosaethyleneglycol] ester. The complementary single stranded oligonucleotides can bind to each other, and phospholipids can be introduced into the lipid bilayer of the cell membrane. Hippocampal neuronal cells from 18 days’ rat embryo and gustatory cells from 2 days’ neonatal mouse were seeded on a poly-D-lysine/laminin and collagen I coated culture slide, respectively. Each complementary DNA-PEG-DPPE was injected to the cultured neuronal cells and gustatory cells, and then the gustatory cells were simply placed on the neuronal cells for DNA-mediated cellular self-assembly. After 1~2 days, assembled neuronal and gustatory cells were treated by bitter taste stimuli and their response were recorded by monitoring the flux of intracellular calcium ions.