HUDSON, T.; PHILLIP, K.; BROWN, K.; NDUKA, E.; CARROLL, M.A.; CATAPANE, E.J.; Medgar Evers College, Brooklyn, NY; Medgar Evers College, Brooklyn, NY; Kingsborough Community College, Brooklyn, NY; Medgar Evers College, Brooklyn, NY; Medgar Evers College, Brooklyn, NY; Medgar Evers College, Brooklyn, NY; ; : Identification and Distribution of Dopamine and Serotonin in Crassostrea virginica by HPLC and ELISA
Biogenic amines are neurotransmitters and hormones in animals. They are well studied in the bivalve mollusc Mytilus edulis, but not as well in oysters, particularly Crassostrea virginica. We studied their presence in C. virginica using an isocratic, ion-pairing HPLC analysis with fluorescence detection to resolved norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, 3,4- dihydroxyphenylalanine, tryptophan, tyrosine, serotonin, 5-hydroxytryptophan, N-acetyl serotonin, tyramine and octopamine. Tissues were dissected, weighed, homogenized, centrifuged, filtered and injected into the HPLC system fitted with a BDS Hypersil C18 column and a Hitachi F-1050 Spectrofluorometer with a 12 µL flow cell. The mobile phase was 50 mM acetate buffer (pH 4.7) with 1 mM EDTA and 1.1 mM SOS, and methanol (85%/15%, v/v). We quantified norepinephrine, epinephrine, dopamine and serotonin in mantle, gill, heart, palps, posterior adductor muscle and visceral ganglia in low amounts (100s of ng/g) which correlates well with published reports of related bivalves. We then employed an ELISA method to corroborate the HPLC results and possibly improve the sensitivity of the assay. ELISA tests for dopamine and serotonin confirmed their presence and the distribution we determined with the HPLC. We believe the study will be an important step in elucidating neurobiological and neuroendocrine functions in C. virginica and of these biogenic amines in general. The work was supported by grants 1R25GM62003 of NIGMS, 0516041071 of NYSDOE, the CUNY Groundworks Program, and 66273-0035 and 66288-0035 of PSC-CUNY. We thank Frank M. Flower & Sons, Inc., Oyster Bay, NY for supplying oysters.
