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Immunohistofluorescence Localization of Biogenic Amine Receptors in Ganglia and Tissues of the Bivalve Mollusc, Mytilus edulis
Author(s) -
Cummings Delilah,
Jacobs Maxine,
Carroll Margaret A,
Catapane Edward J
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.936.2
Subject(s) - mytilus , serotonin , biogenic amine , histamine , 5 ht receptor , dopamine , receptor , chemistry , biology , anatomy , endocrinology , biochemistry , fishery
Biogenic amines serve as neurotransmitters in both sensory and motor systems in a variety of invertebrates. In bivalves, particularly Mytilus edulis, the physiological roles of serotonin and dopamine have been well studied especially with respect to gill and heart. Recently, in the related bivalve Crassostrea virginica , other biogenic amines, including GABA and histamine along with their receptors, were demonstrated to be present, and these amines are being shown to have physiological roles. In light of this we hypothesize that M. edulis would have receptors for the biogenic amines, serotonin, dopamine, histamine and GABA. We tested this using immunohistofluorescence techniques. Primary antibodies to serotonin, dopamine, histamine and GABA receptors, and secondary FITC‐labeled antibodies were purchased from Abcam and Santa Cruz Biotechnology, and used to visualize the presence or absence of these biogenic amine receptors in cerebral ganglia, visceral ganglia, and the animals' innervated gill and mantle. Briefly, tissues were dissected, snap frozen, sectioned on a cryostat, fixed with EDAC (N‐Ethyl‐N′‐(3‐dimethylaminopropyl) carbodiimide hydrochloride), treated with blockers, and incubated with primary, then secondary antibodies. Controls were treated the same way except without primary antibodies. Treated cryostat sections were viewed with a Zeiss epilume fluorescence microscope fitted with a ProgRes C3 Peltier cooled camera, or with a Leica epilume fluorescence microscope fitted with a Leica DFC400 camera. Both microscopes had 100 watt mercury lamps and FITC excitation/emission filters (Ex 470 nm, Em 525 nm). The results show serotonin 2A‐like, dopamine D2‐like, histamine H2‐like and GABA R1‐6‐like receptors present in cerebral ganglia, visceral ganglia, gill and in the sensory tentacles of the mantle rim. This comparative study provides new knowledge of the neurobiology of M. edulis by confirming the presence of these biogenic amine receptors in the animals' nervous system and innervated organs, in a comparable manner to what has been reported in C. virginica . It also provides a foundation to further study the physiological roles for histamine and GABA in M. edulis . Support or Funding Information This work was supported in part by grant 2R25GM06003 of the Bridge Program of NIGMS, grant 690340047 of PSC‐CUNY and a Carnegie Foundation award.