TY - JOUR
T1 - Discovery of leucokinin-like neuropeptides that modulate a specific parameter of feeding motor programs in the molluscan model, Aplysia
AU - Zhang, Guo
AU - Vilim, Ferdinand S.
AU - Liu, Dan Dan
AU - Romanova, Elena V.
AU - Yu, Ke
AU - Yuan, Wang Ding
AU - Xiao, Hui
AU - Hummon, Amanda B.
AU - Chen, Ting Ting
AU - Alexeeva, Vera
AU - Yin, Si Yuan
AU - Chen, Song An
AU - Cropper, Elizabeth C.
AU - Sweedler, Jonathan V.
AU - Weiss, Klaudiusz R.
AU - Jing, Jian
N1 - Funding Information:
This work was supported by National Natural Science Foundation of China Grants 31671097, 31371104, J1103512, and J1210026; NINDS, National Institutes of Health, Grants RO1 NS066587, RO1 NS070583, and RO1 NS031609; NIDA, National Institutes of Health, Grant P30 DA018310; and National Science Foundation Grant CHE 16-067915. The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health and other funding agencies.
Publisher Copyright:
© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2017/11/17
Y1 - 2017/11/17
N2 - A better understanding of neuromodulation in a behavioral system requires identification of active modulatory transmitters. Here, we used identifiable neurons in a neurobiological model system, the mollusc Aplysia, to study neuropeptides, a diverse class of neuromodulators. We took advantage of two types of feeding neurons, B48 and B1/B2, in the Aplysia buccal ganglion that might contain different neuropeptides. We performed a representational difference analysis (RDA) by subtraction of mRNAs in B48 versus mRNAs in B1/B2. The RDA identified an unusually long (2025 amino acids) peptide precursor encoding Aplysia leucokinin-like peptides (ALKs; e.g. ALK-1 and ALK-2). Northern blot analysis revealed that, compared with other ganglia (e.g. the pedal-pleural ganglion), ALK mRNA is predominantly present in the buccal ganglion, which controls feeding behavior.Wethen used in situ hybridization and immunohistochemistry to localize ALKs to specific neurons, including B48. MALDI-TOF MS on single buccal neurons revealed expression of 40 ALK precursor-derived peptides. Among these, ALK-1 and ALK-2 are active in the feeding network; they shortened the radula protraction phase of feeding motor programs triggered by a command-like neuron. We also found that this effect may be mediated by the ALK-stimulated enhancement of activity of an interneuron, which has previously been shown to terminate protraction. We conclude that our multipronged approach is effective for determining the structure and defining the diverse functions of leucokinin-like peptides. Notably, theALKprecursor is the first verified nonarthropod precursor for leucokinin-like peptides with a novel, marked modulatory effect on a specific parameter (protraction duration) of feeding motor programs.
AB - A better understanding of neuromodulation in a behavioral system requires identification of active modulatory transmitters. Here, we used identifiable neurons in a neurobiological model system, the mollusc Aplysia, to study neuropeptides, a diverse class of neuromodulators. We took advantage of two types of feeding neurons, B48 and B1/B2, in the Aplysia buccal ganglion that might contain different neuropeptides. We performed a representational difference analysis (RDA) by subtraction of mRNAs in B48 versus mRNAs in B1/B2. The RDA identified an unusually long (2025 amino acids) peptide precursor encoding Aplysia leucokinin-like peptides (ALKs; e.g. ALK-1 and ALK-2). Northern blot analysis revealed that, compared with other ganglia (e.g. the pedal-pleural ganglion), ALK mRNA is predominantly present in the buccal ganglion, which controls feeding behavior.Wethen used in situ hybridization and immunohistochemistry to localize ALKs to specific neurons, including B48. MALDI-TOF MS on single buccal neurons revealed expression of 40 ALK precursor-derived peptides. Among these, ALK-1 and ALK-2 are active in the feeding network; they shortened the radula protraction phase of feeding motor programs triggered by a command-like neuron. We also found that this effect may be mediated by the ALK-stimulated enhancement of activity of an interneuron, which has previously been shown to terminate protraction. We conclude that our multipronged approach is effective for determining the structure and defining the diverse functions of leucokinin-like peptides. Notably, theALKprecursor is the first verified nonarthropod precursor for leucokinin-like peptides with a novel, marked modulatory effect on a specific parameter (protraction duration) of feeding motor programs.
UR - http://www.scopus.com/inward/record.url?scp=85034580978&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85034580978&partnerID=8YFLogxK
U2 - 10.1074/jbc.M117.795450
DO - 10.1074/jbc.M117.795450
M3 - Article
C2 - 28924050
AN - SCOPUS:85034580978
SN - 0021-9258
VL - 292
SP - 18775
EP - 18789
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 46
ER -