Identification and characterization of the feeding circuit-activating peptides, a novel neuropeptide family of aplysia.
We use a multidisciplinary approach to identify, map, and characterize the bioactivity of modulatory neuropeptides in the circuitry that generates feeding behavior in Aplysia. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of the cerebral-buccal connective (CBC), a nerve containing axons of many interneurons that control feeding behavior of Aplysia, was used to identify neuropeptides that may participate in generation and shaping of feeding motor programs. Using this functionally oriented search, we identified a novel family of peptides that we call the feeding circuit-activating peptides (FCAPs). Two peptides with masses identical to those observed in the CBCs (molecular weight 1387 and 1433) were purified from buccal ganglia and partially sequenced using mass spectrometry. The amino acid sequence was then used to clone the FCAP precursor, which encodes multiple copies of eight different FCAPs. The two FCAPs present in highest copy number correspond to those observed in the CBC. The distribution of FCAP expression was mapped using Northern analysis, whole-mount in situ hybridization, and immunocytochemistry. Consistent with our initial findings, FCAP-immunopositive axons were observed in the CBC. Furthermore, we found that FCAP was present in some cerebral-buccal and buccal-cerebral interneurons. As their name suggests, FCAPs are capable of initiating rhythmic feeding motor programs and are the first neuropeptides with such activity in this circuit. The actions of FCAPs suggest that these peptides may contribute to the induction and maintenance of food-induced arousal. FCAPs were also localized to several other neuronal systems, suggesting that FCAPs may play a role in the regulation of multiple behaviors.
Department of Chemistry and Beckman Institute, University of Illinois, Urbana, Illinois 61801, USA.
J V
JV
Sweedler
L
L
Li
S S
SS
Rubakhin
V
V
Alexeeva
N C
NC
Dembrow
O
O
Dowling
J
J
Jing
K R
KR
Weiss
F S
FS
Vilim
D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia; D001370:Axonal Transport / Q000502:physiology; D001369:Axons / Q000378:metabolism; D015152:Blotting, Northern; D003001:Cloning, Molecular; D005247:Feeding Behavior / Q000502:physiology*; D017952:Ganglia, Invertebrate / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D007395:Interneurons / Q000737:chemistry / Q000502:physiology; D008969:Molecular Sequence Data; D009415:Nerve Net / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology*; D009479:Neuropeptides / Q000032:analysis / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D009928:Organ Specificity; D010507:Periodicity; D016133:Polymerase Chain Reaction; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009479:Neuropeptides; D012333:RNA, Messenger