PubMed1992
The neuroexcitatory peptide isolated from Achatina ganglia was identical to the synthetic Gly-D-Phe-L-Ala-L-Asp with respect to either the bioassay experiments using the Achatina neurones or the instrumental analysis (1H-NMR, SIMS, CD and HPLC). We termed it achatin-I (yield: 50 micrograms from 30,000 animals). Its stereoisomer, Gly-L-Phe-L-Ala-L-Asp, termed achatin-II, was also isolated from the ganglia (yield: 17 micrograms), but this was ineffective on the Achatina neurones. Of the eight possible stereoisomers, only achatin-I markedly showed excitatory effects on the two Achatina neurones, PON and TAN, and [D-Ala3] achatin-I (Gly-D-Phe-D-Ala-L-Asp) had the slight effects. Among the fourteen neurones tested, seven, including the two mentioned above, were excited by achatin-I, whereas no neurone was inhibited. Achatin-I produced an inward current (Iin) with an increase in the membrane conductance (g) under voltage clamp. ED50 of achatin-I for exciting the neurones were 0.20-1.47 x 10(-5) M, and its Emax were 6.33-5.02 nA. Of the achatin-I analogues examined, only the three, Gly-Gly-D-Phe-L-Ala-L-Asp, D-Phe-L-Ala-L-Asp and Gly-D-Phe-L-Ala-L-Asn, produced Iin, but much smaller than that of achatin-I. The equiactive molar ratios (EMRs) of the four effective related peptides (three analogues and a stereoisomer) vs. achatin-I were: 8-60 for Gly-Gly-D-Phe-L-Ala-L-Asp, 200 - > 250 for D-Phe-L-Ala-L-Asp and > 200 for Gly-D-Phe-L-Ala-L-Asn and Gly-D-Phe-D-Ala-L-Asp. The Iin induced by achatin-I was blocked under the /Na+/0-free state, but unaffected under the [Ca2+]-free (replaced with Co2+), [Cl-]0-free or [K+]-enriched (3.0 x) medium, indicating that the Iin is produced by the gNa increase of neuromembrane. We propose that achatin-I having a D-phenylalanine residue is an excitatory neurotransmitter of the Achatina neurones.