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Riboflavin as a source of autofluorescence in Eisenia fetida coelomocytes | JoVE Visualize

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Riboflavin as a source of autofluorescence in Eisenia fetida coelomocytes

Beata Koziol¹, Magdalena Markowicz, Jerzy Kruk

¹Department of Evolutionary Immunobiology, Institute of Zoology, Jagiellonian University, Krakow, Poland.

Photochemistry and Photobiology|January 7, 2006

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Summary

Earthworm eleocytes exhibit autofluorescence due to riboflavin, a vitamin B2 derivative. This finding helps understand the molecular basis of fluorescence in these important immune cells.

Area of Science:

Immunology
Biochemistry
Zoology

Background:

Earthworm coelomocytes, including eleocytes (chloragocytes), possess granules and exhibit autofluorescence.
Previous studies confirmed autofluorescence in eleocytes of various earthworm species using microscopy and flow cytometry.

Purpose of the Study:

To elucidate the molecular origin of autofluorescence in Eisenia fetida eleocytes.
To identify the specific fluorophores responsible for eleocyte autofluorescence.

Main Methods:

Coelomocytes were expelled from Eisenia fetida via dorsal pores using electric shock.
Spectrofluorometry (excitation/emission spectra, fluorescence lifetime) and High-Performance Liquid Chromatography (HPLC) were employed.

Main Results:

Riboflavin was identified as the primary fluorophore responsible for eleocyte autofluorescence.
Flavin mononucleotide (FMN) and flavin-adenine dinucleotide (FAD) were not found to be the main contributors.
Lipofuscins are suggested as potential secondary contributors to the autofluorescence.

Conclusions:

Riboflavin is the principal molecule causing autofluorescence in Eisenia fetida eleocytes.
Understanding these fluorophores aids in studying earthworm immunology and physiology.

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Riboflavin As A Source Of Autofluorescence In Eisenia Fetida Coelomocytes