Identification, physiological actions, and distribution of VYRKPPFNGSIFamide (Val1)-SIFamide) in the stomatogastric nervous system of the American lobster Homarus americanus.
In this study, the peptide VYRKPPFNGSIFamide (Val(1)-SIFamide) was identified in the stomatogastric nervous system (STNS) of the American lobster, Homarus americanus, using matrix-assisted laser desorption/ionization-Fourier transform mass spectrometry (MALDI-FTMS). When bath-applied to the stomatogastric ganglion (STG), synthetic Val(1)-SIFamide activated the pyloric motor pattern, increasing both burst amplitude and duration in the pyloric dilator (PD) neurons. To determine the distribution of this novel SIFamide isoform within the lobster STNS and neuroendocrine organs, a rabbit polyclonal antibody was generated against synthetic Val(1)-SIFamide. Whole-mount immunolabeling with this antibody showed that this peptide is widely distributed within the STNS, including extensive neuropil staining in the STG and commissural ganglia (CoGs) as well as immunopositive somata in the CoGs and the oesophageal ganglion. Labeling was also occasionally seen in the pericardial organ (PO), but not in the sinus gland. When present in the PO, labeling was restricted to fibers-of-passage and was never seen in release terminals. Adsorption of the antibody by either Val(1)-SIFamide or Gly(1)-SIFamide abolished all Val(1)-SIFamide staining within the STNS, including the STG neuropil, whereas adsorption by other lobster neuropeptides had no effect on immunolabeling. These data strongly suggest that the staining we report is a true reflection of the distribution of this peptide in the STNS. Collectively, our mass spectrometric, physiological, and anatomical data are consistent with Val(1)-SIFamide serving as a locally released neuromodulator in the lobster STG. Thus, our study provides the first direct demonstration of function for an SIFamide isoform in any species.
Department of Biology, University of Washington, Seattle, 98195-1800, USA. pdickins@bowdoin.edu
Andrew E
AE
Christie
Elizabeth A
EA
Stemmler
Braulio
B
Peguero
Daniel I
DI
Messinger
Heather L
HL
Provencher
Pieter
P
Scheerlinck
Yun-Wei A
YW
Hsu
Maureen E
ME
Guiney
Horacio O
HO
de la Iglesia
Patsy S
PS
Dickinson
D000818:Animals; D004064:Digestive System / Q000187:drug effects / Q000294:innervation*; D017952:Ganglia, Invertebrate / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D008959:Models, Neurological; D008121:Nephropidae / Q000033:anatomy & histology / Q000378:metabolism*; D009420:Nervous System / Q000378:metabolism*; D009424:Nervous System Physiological Phenomena / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009479:Neuropeptides / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D009490:Neurosecretory Systems / Q000378:metabolism; D011706:Pyloric Antrum / Q000187:drug effects; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013997:Time Factors
D009479:Neuropeptides