Cholecystokinin and gastrin forms in the nervous system.

Cholecystokinin octapeptide is the predominant representative of the gastrin-CCK family in the central nervous system. Other forms occur in low concentrations, or restricted locations, as do the gastrins. The pathways of biosynthetic processing can now be studied in detail, following the elucidation of the cDNA sequence for the two peptides. In the vagus both CCK and gastrin can be found. Brain stem neurons receiving an input from gastric mechanoreceptors respond to CCK-8, but most do not respond to gastrin given intravenously or intra-arterially. This system, which may well be involved in mediating the peripheral satiety effect of CCK, is therefore able to distinguish between the two peptides.

Vertebrate brain-gut peptides related to FMRFamide and Met-enkephalin Arg6Phe7.

Many mammalian brain-gut peptides are known to be represented in invertebrate nervous systems; we have now examined the possibility that an invertebrate neuropeptide occurs in vertebrates. Antisera were raised in rabbits to the molluscan neuropeptide. Phe-Met-Arg-Phe-NH2 (FMRFamide). The antiserum used for radioimmunoassay and immunocytochemistry is highly specific for the C-terminus of the tetrapeptide. In radioimmunoassays of tissue extracts of brain, gut and pancreas of various vertebrates (chicken, frog, dog, rat) concentrations of immunoreactive material up to about 200 pmol/g have been recorded. The immunoreactive material in chicken pancreas behaves on gel filtration and ion exchange chromatography as a molecule that is larger and less basic peptide that FMRFamide. Immunocytochemical studies have demonstrated an endocrine cell origin for FMRFamide-like material in chicken pancreas, and in dog ileum. In brain, FMRFamide can be localised to nerve cell bodies (frog) and nerve fibres (frog and rat). Synthetic FMRFamide has been shown to have excitatory actions on brain stem neurons in the rat. It is suggested that neurons in the rat central nervous system have receptors for FMRFamide that normally bind endogenous material with FMRFamide immunoreactivity.

CNS effects of circulating CCK8: involvement of brainstem neurones responding to gastric distension.

Cholecystokinin octapeptide (CCK8) given i.v. or i.p. produces a variety of behavioural and CNS effects; these actions are probably exerted at a peripheral site but the neuronal pathways involved are uncertain. We show here that i.v. CCK8 acts on neurones in the n. tractus solitarius with an input from the stomach; cells are either excited or depressed by CCK8 and gastric distension, and responses to the two stimuli are always in the same direction. The responses to close arterial injection of CCK8 indicate a site of action within the splanchnic bed, and most probably a direct action on the vagal afferents mediating gastric mechanoreceptor discharge.

An analysis of the responses of rat striatal neurones to scrotal skin temperature.

The responses of neurones in the caudate-putamen complex of anaesthetised rats to different scrotal skin temperatures were examined, together with the electroencephalogram (EEG). Caudate neuronal firing patterns did not change independently of rate, unlike the thermo-responsive cells of the hypothalamus previously reported. The scrotal skin temperature threshold for the caudate neuronal response corresponds precisely with the temperature which provokes desynchronisation of the EEG.

Alteration of the discharge pattern of rat diencephalic neurones with scrotal skin temperature.

Neuronal responses to different scrotal skin temperatures were examined in the hypothalamus of anaesthetised male rats. Mean firing rate and interspike intervals were calculated on-line by microcomputers. Two types of response were observed when the scrotal skin was warmed: an abrupt change in mean firing rate coupled with a change in firing pattern, or a change of pattern unaccompanied by any change in mean rate. These results suggest that hypothalamic cells can convey information independently of their mean firing rate.

The effects of N-methyl-D-aspartate (NMDA) and non-NMDA receptor agonists and antagonists on hypothalamic neurones in the anaesthetized rat which respond to changes in scrotal skin temperature.

There are neurones in the pre-optic anterior hypothalamus of the anaesthetised rat which respond to changes in scrotal skin temperature. Neuronal firing rate can be increased by raising the scrotal skin temperature above 36 degrees C, and also by ionophoretic application of either N-methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) (a non-NMDA receptor agonist). The increase in neuronal firing evoked by thermal stimulation was prevented or diminished by concurrent application of either D-2-amino-5-phosphonopentanoic acid (D-AP5) (an NMDA receptor antagonist) or 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (a non-NMDA receptor antagonist) at doses which were sufficient to abolish or significantly diminish the effect of the specific agonist. It is probable that both NMDA and non-NMDA receptors are involved in the scrotal thermoafferent pathway in the rat.

Mammalian neuronal actions of FMRFamide and the structurally related opioid Met-enkephalin-Arg6-Phe7.

Since the enkephalins were first isolated a number of opioid peptides have been discovered, including a heptapeptide with the sequence Tyr-Gly-Gly-Phe-Met-Arg-Phe (Met-enkephalin-Arg6-Phe7). The heptapeptide was first isolated from chromaffin granules in bovine adrenal medulla, but using immunochemical techniques it has now been identified in human, rat and bovine brains. The C-terminal tetrapeptide of this molecule (Phe-Met-Arg-Phe) occurs in amidated form as the molluscan peptide FMRFamide. Antisera raised against FMRFamide have revealed immunoreactive material in the brains of several vertebrate species, including the rat where it occurs in nerve cell bodies and terminals. I now report that ionophoretically applied FMRFamide has an excitatory effect on rat medullary neurones which is unaffected by the opiate antagonist naloxone. In contrast, Met-enkephalin-Arg6-Phe7 and leucine-enkephalin (Leu-enkephalin) have predominantly depressant effects, which suggests that FMRFamide acts at a separate receptor.

Mechanisms of action of peripherally administered cholecystokinin octapeptide on brain stem neurons in the rat.

We have investigated the pathway and the mechanism by which cholecystokinin octapeptide (CCK-8), given systemically, may influence the discharge of brain stem neurons that have an input from the stomach. Extracellular recordings were made from neurons in the nucleus of the solitary tract (NTS), where vagal afferents terminate, and from neighboring regions of the dorsal medial medulla. Gastric distension and CCK-8 injected intra-aortically close to the stomach evoked either excitatory or inhibitory responses that were abolished by cervical vagal section. In animals from which the celiac/superior mesenteric ganglia were removed, or the gastric antrum resected 2 weeks earlier, responses to gastric distension and CCK-8 were maintained. The effects of CCK-8 are unlikely to be secondary to changes in smooth muscle tone because CCK-8 decreased pressure in the body of the stomach, while distension increased it. Moreover, intravenous noradrenaline and vasoactive intestinal peptide had effects similar to CCK-8 on intragastric pressure, but evoked different patterns of responses from brain stem neurons. The results are consistent with the idea that CCK-8 acts directly on vagal mechanoreceptive endings in the gastric corpus wall. It is well known that peripheral administration of CCK-8 influences short-term regulation of food intake. The effects described here may reflect the pathway by which peripheral CCK influences CNS function.

A novel active pentapeptide from chicken brain identified by antibodies to FMRFamide.

The tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) and peptides structurally related to it, have been isolated from molluscan ganglia. They have widespread actions on both invertebrate and vertebrate tissues and there is increasing evidence that they are an important group of invertebrate peptide neurotransmitters. It is of interest that the primary amino acid sequence of FMRFamide forms the C-terminal tetrapeptide of an enkephalin-like heptapeptide (Met-enkephalin-ArgPhe) isolated from bovine adrenal medulla and striatum. Antisera to FMRFamide have been shown to react in radioimmunoassay and immunohistochemistry with material in the central nervous system of various vertebrate species, but the identity of this material, and in particular its relationship to the opioid heptapeptide, remains uncertain. We have used antibodies specific for the C-terminus of FMRFamide in radioimmunoassays to monitor purification of the material in chicken brain. We describe here the sequence of one of the peptides obtained. It is a biologically active peptide which does not seem to be related to other known vertebrate neuropeptides.