Fluorescence of plant microspores as biosensors

Possibilities of fluorescent microscopic single-cell analysis on plant microspores as biosensors for study of chemosignaling involving neurotransmitters and mechanisms of action of fluorescent medicinal compounds—antagonists of neurotransmitters were studied on some examples. By methods of luminescence microscopy, microspectrofluorimetry and laser scanning confocal microscopy using as an example field horsetail Equisetum arvense microspores, the penetration of these compounds into the cell and associate it with individual compartments (estimated as the changes in their autofluorescence) has been analyzed. Fluorescent in blue antagonists of neurotransmitters d-tubocurarine, yohimbine and azulene (blockers of cholinoreceptor, adrenoreceptor, and histamine receptor, respectively) decreased the number of the E. arvense cells with red fluorescence. Tubocurarine and yohimbine bound to the cellular surface and did not penetrate into the cells. Azulene was found both on the cell surface and inside cells, demonstrating blue (excitation 360–380 nm) or green (excitation 420 nm) fluorescence of DNA-containing organelles. The effects of lipophilic (lecithin and amphotericin B) and proteinous (albumin, enzyme cholinesterase, cytoskeleton proteins actin, myosin, and titin) compounds on the manifestation of the effects of the neurotransmitters and antagonist d-tubocurarine have been shown. The intensity of the red light at 680 nm, has evolved in many variants. Most notable was the decline of the emission in the presence of albumin and cholinesterase as compared with the action of dopamine itself. After the addition of the cytoskeleton proteins and cholinesterase to the medium, the decrease of red fluorescence intensity, usually induced by d-tubocurarine, was not observed.