neuropeptide-y and galanin-(1-13)-spantide-amide

Galanin is a neuropeptide that causes a marked pressor response in several non-mammalian vertebrate species, and some marsupials. In this study, the effect of three galanin antagonists were tested on the pressor response to an intravenous dose (6.3 nmol/kg) of porcine galanin in anaesthetised Cane toads, Bufo marinus. Antagonists were injected at either 20 or 50 times the molar dose (x MD) of galanin. The antagonist, C7 (Galanin 1-13-spantide) reduced the pressor effect of galanin by 32.2 +/- 6.0% when delivered at 20 x MD (n = 4) and by 42.9 +/- 15.7% when delivered at 50 x MD (n = 4) of galanin, the response recovering within 30 min. A second antagonist, M32a (Galanin 1-13-NPY 24-36) had no effect on the pressor response to galanin at 20 x MD (n = 4), but significantly reduced the pressor effect by 54.8 +/- 6.4% at 50 x MD (n = 5), which also recovered within 30 min. Administration of a third antagonist, galantide or M15 (Galanin 1-13-Substance P5-11), resulted in a profound drop in blood pressure, and did not affect the response to galanin at either dose. In conclusion, C7 and M32a are effective, short-term antagonists of the blood pressure effects of galanin in the toad.

Topics: Animals; Blood Pressure; Bufo marinus; Galanin; Heart Rate; Neuropeptide Y; Peptide Fragments; Recombinant Fusion Proteins; Substance P; Swine; Vasoconstrictor Agents

1. The role of endogenous galanin in somatosensory processing has been studied with galanin receptor antagonists. The new galanin receptor ligands C7, M32, M38 and M40 bind with high affinity (Kd in nanomolar range) to spinal cord galanin receptors and possess oxidative stability as compared to earlier generations of peptide ligands. These peptides have been examined in the spinal flexor reflex model where exogenous galanin exhibited biphasic excitatory and inhibitory effects. 2. Intrathecal administration of C7 [galanin(1-13)-spantide] and M32 [galanin (1-13)-neuropeptide Y(25-36) amide] blocked facilitation of the nociceptive flexor reflex induced by 30 pmol intrathecal galanin in decerebrate, spinalized rats in a dose-dependent manner, thus behaving as antagonists of the galanin receptor. In contrast, M38[galanin(1-13)-(Ala-Leu)3-Ala amide] and M40 [galanin(1-13)-Pro-Pro-(Ala-Leu)2-Ala amide], exhibited only weak antagonism at high doses in this model. Moreover, lower doses of M40 potentiated galanin-induced reflex facilitation. C7 was neurotoxic at high doses in the rat spinal cord. 3. M32 and C7 were potent antagonists of galanin receptors in rat spinal cord, in correlation with their in vitro binding characteristics. In contrast, M38 and M40, despite their high in vitro affinity, exhibited only very weak antagonism. Moreover, M40 may also behave as a partial agonist. 4. Previous studies have shown that the galanin receptor may be heterogeneous. The discrepancy between in vitro binding and in vivo antagonistic potency of M38 and M40 may also suggest the presence of different galanin receptor subtypes within the rat spinal cord. However, other explanations for the discrepancy, such as differences in metabolic stability, diffusion rates and penetration to the site of action are also possible.

Topics: Amino Acid Sequence; Animals; Binding, Competitive; Female; Galanin; Injections, Spinal; Molecular Sequence Data; Neuropeptide Y; Peptide Fragments; Peptides; Rats; Rats, Sprague-Dawley; Receptors, Galanin; Receptors, Gastrointestinal Hormone; Recombinant Fusion Proteins; Spinal Cord; Substance P