Elsevier

Peptides

Volume 19, Issue 3, 1998, Pages 487-493
Peptides

Articles
Identification and Characterization of a Myomodulin-Like Peptide in the Leech

https://doi.org/10.1016/S0196-9781(97)00419-1Get rights and content

Abstract

Wang, Y., D. A. Price and C. L. Sahley. Identification and characterization of a myomodulin-like peptide in the leech. Peptides 19(3) 487–493, 1998.—A novel myomodulin-like peptide, GMGALRLamide, has been purified and sequenced from extracts of 1000 medicinal leech nerve cords. Synthetic leech myomodulin-like peptide blocked the specific staining pattern of leech ganglia by the antiserum against Aplysia myomodulin A PMGMLRLamide. Moreover, the synthetic leech myomodulin-like peptide GMGALRLamide showed identical neuronal modulation effect on the giant leech Retzius cell compare to that by the synthetic Aplysia myomodulin A PMGMLRLamide.

Section snippets

Animals

Adult leeches, Hirudo medicinalis (4–6 g) were obtained from Leeches USA (Westbury, NY). Leeches were maintained in artificial leech pond water [0.5 g of Hirudo salt (Leeches USA) dissolved in 1 liter of dH2O] at 15°C with 12 h/12 h light/dark cycle.

Reagents

Synthetic Aplysia myomodulin-A PMSMLRL-NH2 was purchased from Peninsula Lab (Belmont, CA). Leech myomodulin-like peptide GMGALRL-NH2 was synthesized by Research Genetics (Huntsville, AL). The antisera for RIA and immunocytochemistry were generous

Identification of Myomodulin-Like Immunoreactivity by HPLC

The fractions resulting from the Sep-Pak separations of the ten batches of 100 dissected leech nerve cords were assayed for immunoreactivity and all the active fractions were pooled. After five sequential steps of reverse-phase HPLC (Fig. 1), two fused absorbance (210 nm) peaks were observed with the immunoreactivity peak associated with the first peak (indicated by arrowhead in Fig. 2A). Fraction 63 (indicated by arrowhead in Fig. 2B) was analyzed by automated sequencing and MALDItof mass

Discussion

We have isolated a novel myomodulin-like heptapeptide GMGALRL-NH2 from the leech, Hirudo medicinalis. The majority of the myomodulin family peptides identified so far are from molluscs and their effects appear to be modulation of neuromuscular transmission 6, 12, 26, 28, 29. The results presented here open up the possibility that myomodulin peptides are also involved in neuronal modulation underlying various animal behaviors.

In our final HPLC step, the immunoreactivity was associated with the

Acknowledgements

We thank Drs. J. Strong, S. Rossie, D. Ready and A. Bieber for critical reading of the manuscript. This work was supported by NSF Grant BIR 9512962 and NIMH Grant R01MH44789 (to C.L.S.).

References (31)

  • B.D. Evans

    The effects of myomodulin and structurally related neuropeptides on skeletal neuromuscular transmission in the locust

    J. Exp. Biol.

    (1994)
  • M.B. Feany

    Neuropeptide modulation of learning and memory processes

    Rev. Neurosci.

    (1996)
  • M. Fujiwara-Sakata et al.

    Neuropeptides regulate cardiac activity of a prosobranch mollusc, Rapana thomasiana

    Cell Tissue Res.

    (1992)
  • Greenberg, M. J.; Doble, K. E.; Lesser, W.; Lee, T. D.; Pennell, N. A.; Morgan, C. G.; Price, D. A. Characterization of...
  • Ikeda, T.; Minakata, H.; Fujita, T.; Muneoke, Y.; Kiss, T.; Hiripi, L.; Nomoto, K. Neuropeptides isolated from Helix...
  • Cited by (18)

    • Annelids Neuro-Endrocrino-Immune Response

      2024, Frontiers in Invertebrate Physiology: A Collection of Reviews: Volume 3: Annelida and Echinodermata
    View all citing articles on Scopus
    View full text