An additional arginine-vasotocin-related peptide, vasotocinyl-Gly-Lys, in Xenopus neurohypophysis.

The neurohypophysis of Xenopus and that of Ranidae and Bufonidae contain hydrin 1 (vasotocinyl-Gly-Lys-Arg) and hydrin 2 (vasotocinyl-Gly), respectively. In order to test the aldosterone-releasing activity of arginine vasotocin (AVT) and hydrin 1, purification of these peptides from an acid-extract of the neurointermediate lobe of Xenopus laevis was performed using an ODS-silica cartridge and reverse-phase and ion-exchange HPLC columns. As a result, an additional AVT-related peptide was newly found. Amino-acid analysis revealed that this peptide is vasotocinyl Gly-Lys (AVT-GK). The aldosterone-releasing activity of AVT-GK was equivalent to that of hydrin 1 (AVT-GKR) and lower than that of AVT. Like AVT and AVT-GKR, AVT-GK were effective in stimulating water flux from the isolated urinary bladder of the toad. Since AVT-GK is regarded as an intermediate between hydrin 1 and hydrin 2 in terms of its C-terminal form, it was designated hydrin 1'.

Arginine vasotocin (AVT) and AVT-related peptide are major aldosterone-releasing factors in the bullfrog neurointermediate lobe.

Two major components which stimulate aldosterone release from Xenopus adrenocortical tissue were isolated from an acid-acetone extract of the neurointermediate lobes of the bullfrog (Rana catesbeiana) using C18 Sep-Pak cartridges, Sephadex G-50, and reverse-phase HPLC columns. One of the components was identified as arginine vasotocin (AVT) from its HPLC profile and amino acid sequence analysis. The other was an AVT-like decapeptide with an extra glycine residue at the C-terminus of nonamidated AVT, which was recently termed hydrin 2. The yields of these two peptides were almost the same. They also showed equipotent activity in stimulating water flux from the isolated urinary bladder of the toad (Bufo japonicus).

Evolutionary specificity of hydrins, new hydroosmotic neuropeptides: occurrence of hydrin 2 (vasotocinyl-Gly) in the toad Bufo marinus but not in the viper Vipera aspis.

Hydrin 2 (vasotocinyl-Gly), a hydroosmotic peptide resulting from differential processing of provasotocin and recently identified in frog neurohypophysis, has been looked for in the pituitary gland of an exotic toad (Bufo marinus) and of a reptile (Vipera aspis). Hydrin 2 has been found in the amphibian but not in the reptile. This result confirms the evolutionary specificity of hydrin 2 that has been identified only in frogs and toads but not in birds and reptiles. Occurrence of hydrin 2 is explained by its regulatory function on the water permeability of the skin of anurans.

Hydrins, hydroosmotic neurohypophysial peptides: osmoregulatory adaptation in amphibians through vasotocin precursor processing.

From neurointermediate pituitary glands of Xenopus laevis and Rana esculenta, previously unreported peptides termed hydrins, active on water permeability of frog urinary bladder and frog skin (Brunn or "water-balance" effect), have been isolated and sequenced. These peptides seem to be derived from the pro-vasotocin-neurophysin precursor. Hydrin 1, found in Xenopus, has been identified as vasotocin C-terminally extended with the Gly-Lys-Arg sequence; hydrin 2, found in Rana, has been identified as vasotocin C-terminally extended with glycine. Hydrin 2 has been detected in several Ranidae (R. esculenta, Rana temporaria, Rana pipiens) and Bufonidae (Bufo bufo, Bufo ictericus) and appears to have a large distribution in terrestrial or semiaquatic anurans. Hydrins, in contrast to vasotocin, are not active on rat uterus or rat blood pressure. They are absent from other vasotocin-bearers such as birds and could be involved specifically in water-electrolyte regulation of amphibians.

Arginine vasotocin from the pituitary gland of the lamprey (Petromyzon marinus): isolation and amino acid sequence.

Arginine vasotocin (AVT) was isolated from extracts of sea lamprey pituitary glands (Petromyzon marinus). The amino acid sequence Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Arg-Gly-NH2 is identical to the molecule isolated from teleosts and tetrapods. The total yield was estimated to be 9.6 pmol per gland. No evidence could be found for the existence of a second neurohypophyseal nonapeptide in the lamprey pituitary.

Mesotocin and vasotocin, two neurohypophysial hormones in the ostrich, Struthio camelus.

Two neurohypophysial hormones have been isolated from an avian species, the ostrich, Struthio camelus. Both have been characterized by amino acid analysis and sequence determination. The data obtained suggest that the oxytocin-like hormone is [Ile8-oxytocin] (mesotocin) and the vasopressin-like hormone is [Ile3-vasopressin] (vasotocin). Bioactivity measurements based on urinary conductivity showed vasotocin to be about five times as active as mesotocin.

Precursors of mesotocin and vasotocin in birds: identification of VLDV- and MSEL- neurophysins in chicken, goose, and ostrich.

Precursors of neurohypophysial hormones are small proteins processed into nonapeptide hormones and neurophysins during axonal transport to the neurohypophysis. In mammals, oxytocin is associated with VLDV-neurophysin and vasopressin with MSEL-neurophysin. In birds, mesotocin and vasotocin are found instead of mammalian oxytocin and vasopressin. From goose, chicken and ostrich posterior pituitary glands, two types of neurophysins related to mammalian VLDV- and MSEL-neurophysins, respectively, have been identified by their N-terminal sequences. It is assumed that, as in mammals, hormonal peptide and the first 9 residues of the corresponding neurophysin are encoded by a common exon and that mesotocin and vasotocin, evolutionary predecessors of oxytocin and vasopressin, are associated in the precursors with VLDV-neurophysin and MSEL-neurophysin, respectively.

Pineal peptides.

In pineal extracts several activities have been observed which cannot be explained on the basis of their content of melatonin or other pineal indoles to which an antigonadotropic activity is ascribed. The significance of structurally identified and unidentified substances is not fully understood. The list includes a neurohypophysial peptide, vasotocin (AVT), hypothalamic releasing factors and certain unidentified substances including the antigonadotropin under investigation in our laboratory. Our initial experiments demonstrated the presence of a non-melatonin antigonadotropin (PAG) in partially purified extracts of bovine, ovine, rat and human pineals. Purification by ion exchange chromatography and amino acid analyses have shown that this substance is not AVT. Recent purification studies include paper chromatography and high performance chromatography as final steps. It is concluded that if the active principle in a peptide, or contains a peptide moiety important for the biological activity, it may be present only in minute amounts in the puriest fractions derived and the quantities of recoverable material may be similar to those amounts of releasing factors recoverable from hypothalamic tissue. It is anticipated that large scale preparative methods will be required for structural determination. High performance chromatography may prove to be extremely useful in future studies.