Transient expression of somatostatin sst2 receptors in rat cerebellar nuclei during development.

Adult rat cerebellar nuclei contain a single population of [125I][Leu8,D-Trp22,Tyr25]somatostatin-28 binding sites characterized as sst1 receptors. In the present study, we have investigated the evolution of somatostatin receptors in rat cerebellar nuclei during development by means of quantitative autoradiography on tissue sections. The binding of [125I][Leu8,D-Trp22,Tyr25]somatostatin-28, observed in the primordium of the medial cerebellar nuclei at embryonic day 17, reached a maximum at postnatal day 7 or 10 in the different nuclei. Thereafter, the density of binding sites gradually decreased to the adult level. Competition studies were performed using the somatostatin analogues CH-288 and MK-678 as specific sst1 and sst2 ligands, respectively. Partial inhibition of the radioligand binding by CH-288 and MK-678 revealed the presence of a predominant population of sst1 from embryonic day 19-28 day postnatal and a minor population of sst2 receptors. The use of [125I]MK-678 as a radioligand confirmed the presence of a transient population of sst2 receptors, suggesting that somatostatin could act on rat cerebellar nuclei via sst1 and/or sst2 receptors during development.

des-AA-1,2,5[D-Trp8, IAmp9]somatostatin-14 allows the identification of native rat somatostatin sst1 receptor subtype.

Somatostatin exerts multiple activities by interacting with at least five different receptor subtypes (sst[1-5]). The affinity of des-AA(1,2,5)-[D-Trp8, IAmp9]somatostatin-14 (CH-275) was studied by competition experiments using the non-selective radioligand [125I][Leu8, D-Trp22, Tyr25]somatostatin-28 in areas of the rat brain and pituitary known to express identified receptor subtypes. In the cerebellar nuclei and cerebral cortex, which possess the somatostatin sst1 receptor subtype, CH-275 exhibited a moderate affinity (IC50: 10-50 nM). Conversely, in the hippocampus, immature cerebellum and pituitary which contain different subsets of receptors mRNAs (sst[2-5]), the IC50 values were > 1 microM. These data indicate that CH-275 is an appropriate ligand for the identification of native rat somatostatin sst1 receptor subtype.

Occurrence of two somatostatin variants in the frog brain: characterization of the cDNAs, distribution of the mRNAs, and receptor-binding affinities of the peptides.

In tetrapods, only one gene encoding a somatostatin precursor has been identified so far. The present study reports the characterization of the cDNA clones that encode two distinct somatostatin precursors in the brain of the frog Rana ridibunda. The cDNAs were isolated by using degenerate oligonucleotides based on the sequence of the central region of somatostatin to screen a frog brain cDNA library. One of the cDNAs encodes a 115-amino acid protein (prepro-somatostatin-14; PSS1) that exhibits a high degree of structural similarity with the mammalian somatostatin precursor. The other cDNA encodes a 103-amino acid protein (prepro-[Pro2, Met13]somatostatin-14; PSS2) that contains the sequence of the somatostatin analog (peptide SS2) at its C terminus, but does not exhibit appreciable sequence similarity with PSS1 in the remaining region. In situ hybridization studies indicate differential expression of the PSS1 and PSS2 genes in the septum, the lateral part of the pallium, the amygdaloid complex, the posterior nuclei of the thalamus, the ventral hypothalamic nucleus, the torus semicircularis and the optic tectum. The somatostatin variant SS2 was significantly more potent (4-6 fold) than somatostatin itself in displacing [125I-Tyr0, D-Trp8] somatostatin-14 from its specific binding sites. The present study indicates that the two somatostatin variants could exert different functions in the frog brain and pituitary. These data also suggest that distinct genes encoding somatostatin variants may be expressed in the brain of other tetrapods.

Pharmacological characterization of somatostatin receptors in rat cerebellar nuclei.

Rat cerebellar nuclei contain somatotropin release-inhibiting factor (SRIF) receptors that bind [125I][Leu8,D-Trp22,Tyr25]SRIF-28 but do not bind [125I][Tyr0,D-Trp8]SRIF-14. The aim of the present study was to investigate the pharmacological profile of these receptors by means of binding experiments on tissue sections and quantitative autoradiography. Competition experiments indicated the presence of a single class of [125I][Leu8,D-Trp22,Tyr25]SRIF-28 binding sites in the lateral cerebellar nuclei, showing similar affinities for SRIF-14 and SRIF-28, but low affinity for short-chained analogs. The IC50 values for somatostatin analogs to compete with the binding of [125I][Leu8,D-Trp22,Tyr25]SRIF-28 in the lateral cerebellar nuclei ranked as follows: [Leu8,D-Trp22,Tyr25]SRIF-28 approximately SRIF-14 approximately SRIF-28 < CGP 23996 < D-Phe-Phe-Phe-D-Trp-Lys-Thr-Phe-Thr-NH2 (BIM 23052) < SMS 201-995 approximately N-Ahep-(7-10)SRIF-14-Bzl << MK 678 < D-Phe-Phe-Tyr-D-Trp-Lys-Val-Phe-D-Nal-NH2 (BIM 23056) < D-Phe-c[Cys-Tyr-D-Trp-Lys-Abu-Cys]Nal-NH2 (NC 8-12). Optimum binding of [125I][Leu8,D-Trp22,Tyr25]SRIF-28 did not require divalent cations, and was partly inhibited by guanosine 5' triphosphate. It appears from this study that the rat lateral cerebellar nuclei contain a pure population of receptors exhibiting the same binding characteristics as the recently cloned sstr1 somatostatin receptor. These nuclei could thus provide a useful model in which to investigate the characteristics of native sstr1.

Immunocytochemical evidence for the presence of Met-enkephalin and Leu-enkephalin in distinct neurons in the brain of the elasmobranch fish Scyliorhinus canicula.

Immunohistochemical methods have been used to investigate the distribution of various opioid peptides derived from mammalian proenkephalin in the central nervous system of Scyliorhinus canicula. The results indicate that both Leu- and Met-enkephalin-immunoreactive peptides are present in the dogfish brain. In contrast, enkephalin forms similar to Met-enkephalin-Arg-Phe or Met-enkephalin-Arg-Gly-Leu, and mammalian alpha-neo-endorphin, dynorphin A (1-8), dynorphin A (1-13), and dynorphin A (1-17) were not detected. Met- and Leu-enkephalin immunoreactivities were found in distinct neurons of the telencephalon and hypothalamus. In particular, cell bodies reacting only with the Met-enkephalin antiserum were localized in the preoptic nucleus and in the suprachiasmatic region of the hypothalamus. Conversely, cell bodies reacting only with the Leu-enkephalin antiserum were localized in the pallium and the nucleus lobi lateralis hypothalami. Several areas of the telencephalon and diencephalon exhibited both Met- and Leu-enkephalin-like immunoreactivity, but the two immunoreactive peptides were clearly contained in distinct perikarya. The overall distribution of Met-enkephalin-immunoreactive elements in the dogfish exhibited similarities to the distribution of proenkephalin-derived peptides previously reported for the brain of tetrapods. The fact that Met- and Leu-enkephalin-like peptides were detected in distinct neurons, together with the absence of dynorphin-related peptides, suggests the existence of a novel Leu-enkephalin-containing precursor in the dogfish brain.

Variables processing in expert system building: application to the aetiological diagnosis of infantile meningitis.

We describe a method for the analysis and quantitative synthesis of information in a clinical data file using as an example the aetiological diagnosis of infantile meningitis. The results obtained together with the qualitative knowledge of experts in the field have enabled an experimental system to be constructed. This is performing sufficiently well to be introduced into a hospital environment to assist young doctors facing an emergency situation.