Biomedical subjects

R Benoit

Publications and source records attributed to R Benoit.

At least 73 records · Page 4Linked to original sources

PubMed1985

Immunocytochemical localization of somatostatin-28 in the rat hypothalamus.

Somatostatin-28 (SS28) is considered as a precursor of somatostatin-14 (SS14) and also of the remaining NH2 terminus of SS28, the dodecapeptide SS28 which has been recently characterized. In order to study the cellular and subcellular localization of SS28 in the rat hypothalamus, we have conducted a light and electron microscopic immunocytochemical study, using both the peroxidase-antiperoxidase and the immunogold technique. Several antisera which selectively recognize one or more of these 3 somatostatin-related peptides were used. In serial paraffin sections, it was observed that SS28 was contained in the same neuronal cell bodies which also contain SS28. These neurons were exclusively located in the periventricular nucleus. All the antisera used also produced a strong staining in the external zone of the median eminence. At the electron microscopic level, the 3 peptides were exclusively localized in all the dense core vesicles in cell bodies and axons in the periventricular nucleus and terminals in the median eminence. These results strongly suggest that SS28 and SS14 originate from the precursor SS28 and that processing of the precursor occurs in the cell body. They also support the hypothesis that the 3 peptides are released simultaneously under appropriate stimulation.

Animals↗

PubMed1985

Somatostatin-28 [1-12]-like peptides.

The search for a peptide corresponding to the NH2-terminus of somatostatin-28 (SS-28) in tissues has led to the isolation and characterization of somatostatin-28[1-12] from pancreas and hypothalamus. Somatostatin-28[1-12]-like immunoreactivity [SS-28 [1-12]-LI] is widely distributed throughout the central nervous system and the digestive system of rodents and primates, reaching levels comparable to those of somatostatin-14 (SS-14). Antibodies directed against the C-terminal end of the dodecapeptide are more specific and constitute excellent markers for the "prosomatostatin" system in mammalian tissues. In rat brain, SS-28[1-12]-LI material is highly concentrated in nerve fibres and terminals, especially in the median eminence, layer I of neocortex, the outer molecular layer of the dentate gyrus and the striatum. Additionally, immunoreactivity is observed in large multipolar or occasionally pyramidal-like neurons of the neocortex. SS-28[1-12] is secreted from hypothalamus and amygdala by a calcium dependent mechanism. No biological role is presently known for the dodecapeptide. Two other peptides of Mr = 8000 (8 K) and Mr = 5000 (5 K) which contain SS-28[1-12] at their carboxy-termini are present in acid extracts from rat pancreas, brain and spinal cord. These two peptides were isolated from an acid extract of rat brains using ion-exchange chromatography, gel permeation chromatography and reverse-phase HPLC. Results from amino acid analysis and partial sequencing were compared to the sequence of the cDNA encoding rat pre-prosomatostatin (prepro-SS) and revealed that the 8 K peptide is a 76 amino acid molecule corresponding to prepro-SS[25-100] and that the 5K peptide, which contains 44 amino acids, corresponds to prepro-SS [57-100]. The 5 K peptide was generated after cleavage of a Leu-Leu bond at position 56-57 of prepro-SS. The four most predominant peptides of the "prosomatostatin system" presently characterized are: SS-14, SS-28[1-12], SS-28 and prepro-SS[25-100]. Studies on pooled perfusates from rat hypothalamic tissue show that prepro-SS[25-100] is released with SS-28[1-12] in vitro and accounts for 22% of the total SS-28[1-12]-like immunoreactive material released during depolarization. The 5 K peptide is apparently not secreted. The presence of prepro-SS[25-100] in brain implies that, first, prosomatostatin can serve as an immediate precursor for SS-14 without going through SS-28 as an intermediate step and second, other peptides could conceivably be derived from the cryptic portion of the precursor.

Amygdala↗

PubMed1985

[Embryogenesis of the styloglossus muscle in the human embryo. Its relationship with neighboring structures].

Histogenesis of the muscles of the human tongue has been studied from the 15th week in utero onward: however their intrinsic or extrinsic origin is uncertain. The intrinsic musculature has sometimes be questioned (Saban, 1971). The embryogenesis of the styloglossus muscle has been studied in ten 39 to 58 day old human embryos. Its differentiation beyond the first lingual swellings in contact with a branch of the hypoglossus nerve allows its development to be followed outside the territories of the branchial arches. This study showed its progressive development, from day 42 onward, along bundles of the ceratoglossus and basioglossus muscles. The styloglossus muscle penetrates slowly on day 45 in the transverse fibres located behind the tongue. These latter differentiate earlier, independently and at the same time as the geniohyoid and genioglossus muscles, near the neural extensions of the hypoglossus nerve: they are very rapidly intermixed with the fibres of the genioglossus muscle: the other muscular fibres and those of the styloglossus muscle penetrate only later and progressively. In the current state of knowledge of axial neural organization, it can be concluded that the lingual muscles differentiate from the same type of cells recognized and innervated by the neurons of the hypoglossus nerve. The epigenesis of the different muscles is related to the appearance of their various functions.

Cell Differentiation↗

PubMed1984

Neuropeptides derived from prosomatostatin that do not contain the somatostatin-14 sequence.

Two neuropeptides reacting with antibodies directed against the C-terminal region of somatostatin 28(1-12) were purified to homogeneity from rat brain extracts. Amino acid analysis revealed that the larger peptide (8 kdaltons) consisted of 76 amino acids. Microsequencing established its aminoterminal structure as: Ala-Pro-Ser-Asp-Pro-Arg-Leu-Arg-Gln-Phe-X-Gln-Lys. The 8 kdalton somatostatin 28(1-12)-like peptide corresponds to the whole prosomatostatin molecule without Arg-Lys-somatostatin-14. The smaller peptide (5 kdaltons) consists of 44 amino acids and is generated after cleavage of a Leu-Leu bond at position 56-57 of pre-prosomatostatin. Both 8 kdalton and 5 kdalton somatostatin 28(1-12)-like peptides contain somatostatin 28(1-12) at their C-termini. The 4 most abundant neuropeptides derived from pre-prosomatostatin (pre-proSS) and presently characterized are: somatostatin-14, somatostatin 28(1-12), somatostatin 28 and pre-proSS25-100.

Amino Acid Sequence↗

PubMed1984

Isolation and characterization of the bovine hypothalamic corticotropin-releasing factor.

A 41 amino acid peptide with high intrinsic corticotropin-releasing activity was isolated from 1000 bovine hypothalami by means of immunoaffinity chromatography, gel filtration, and two steps of reverse phase HPLC. The primary structure of the amino terminal 39 amino acids was characterized by gas phase sequence analysis. The sequence of the amidated carboxyl terminal dipeptide was established by digestion of the intact natural product with Staphylococcus aureus V8 protease, dansylation of the digest and comparative reverse phase liquid chromatography studies with the synthetic dansylated dipeptides Ile-Ala-NH2, Ile-Ala-OH, Ala-Ile-NH2 and Ala-Ile-OH. The complete structure of the bovine corticotropin-releasing factor was established as: Ser-Gln-Glu-Pro-Pro-Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val- Leu- Glu-Met-Thr-Lys-Ala-Asp-Gln-Leu-Ala-Gln-Gln-Ala-His-Asn-Asn-Arg-Lys-Leu- Leu- Asp-Ile-Ala-NH2 using approximately 650 pmol of material.

Amino Acid Sequence↗

PubMed1984

The distribution and morphological characteristics of the intracortical VIP-positive cell: an immunohistochemical analysis.

Using a sensitive immunohistochemical procedure, we have undertaken a detailed morphological characterization of the vasoactive intestinal peptide-positive (VIP-positive) neuron in the cerebral cortex of the rat. VIP-positive neurons are present in all regions of cortex, and are usually strongly bipolar, possessing long, radially directed processes with very limited branching in the tangential plane. The most extensive dendritic branching occurs in layers I and deep IV-superficial V, and the density of axonal varicosities is highest in layers II-IV. In the visual cortex, approximately 50% of the labeled cell bodies are in layer II and III and 80% of the labeled cell bodies are contained in layers I-IV (superficial 600 microns of cortex). In order to determine the density and 3-dimensional distribution pattern of these cells, we prepared serial tangential sections through the rat visual cortex, mapped the distribution of all labeled cells in each section on transparent acetate sheets, and compressed these superimposed maps. This analysis demonstrated that: (1) approximately 1% of the cortical neurons are VIP-positive, (2) their distribution is fairly uniform and statistically random, (3) there are no large areas (i.e. with a diameter greater than 100 microns) that lack a VIP-positive cell, (4) on the average, there is one VIP-positive cell per column of 30 microns diameter, and (5) the average nearest neighbor distance on the compressed display is 15 microns. Given the morphological characteristics of VIP-positive cells, these data indicate that each VIP-containing cell is identified with a unique radial volume, which is generally between 15 and 60 microns in diameter, and overlaps with the contiguous domains of neighboring VIP-positive cells. These morphological data support the notion that VIP-containing neurons play an important functional role within radially oriented columns of cerebral cortex.

Animals↗

PubMed1984

Subcellular distribution of somatostatin-14, somatostatin-28 and somatostatin-28 (1-12) in rat brain cortex and comparisons of their respective binding sites in brain and pituitary.

Subcellular distribution and binding characteristics of the three endogenous peptides somatostatin-14 (SRIF-14), somatostatin-28 (SRIF-28) and somatostatin-28(1-12) (SRIF-28(1-12] derived from preprosomatostatin were investigated in the rat brain cortex. The three peptides are predominantly recovered from a crude mitochondrial pellet (P2), containing the pinched off nerve endings. Specific high affinity binding sites for 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 are present on pituitary and brain membranes. Under the same conditions, 125I-N-Tyr-SRIF-28(1-12) binding is undetectable. Moreover, SRIF-28(1-12) does not displace 125I-N-Tyr-SRIF-14 or 125I-N-Tyr-SRIF-28 binding. SRIF-28 is more potent than SRIF-14 to displace 125I-N-Tyr-SRIF-28 binding to brain and pituitary membranes, while both peptides are equipotent to displace 125I-N-Tyr-SRIF-14 binding. Finally, the regional distribution of 125I-N-Tyr-SRIF-14 and 125I-N-Tyr-SRIF-28 binding sites in the brain is identical. In conclusion, the present results are consistent with a neurotransmitter and neurohormonal role for SRIF-14 and SRIF-28. The function of SRIF-28(1-12) in brain remains to be elucidated. Additionally, a differential role for SRIF-14 and SRIF-28 both in adenohypophysis and brain cannot be ascertained at the present time.

Animals↗

PubMed1984

Somatostatin-28(1-12)-like immunoreactive substance is secreted into hypophysial portal vessel blood in the rat.

Somatostatin-28(1-12)-like immunoreactivity was measured in extracts of rat hypophysial portal blood and peripheral blood. The concentration of somatostatin-28(1-12) was higher in portal than in peripheral extracts, and its release into portal vessel blood was increased 4- to 5-fold by electrical stimulation of the median eminence. These results show that somatostatin-28(1-12) may be a physiological neurohormone and/or neurotransmitter.

Animals↗

PubMed1984

Evidence for selective release of somatostatin-14 and somatostatin-28(1-12) from rat hypothalamus.

Cysteamine administration to rats results in a marked depletion of hypothalamic somatostatin-14 (SS14) and a decrease of the potassium-evoked in vitro release of SS14 without a significant change in the content or release of somatostatin-28(1-12)-like immunoreactivity (SS28(1-12)-L1). Furthermore, cysteamine enhances the spontaneous release and markedly potentiates the potassium-evoked release of SS14 in the in vitro slice preparation. However, in vitro-administered cysteamine does not alter the spontaneous or potassium-evoked release of SS28(1-12)-LI. Immunohistochemical visualization of hypothalamic neuronal cell bodies and fibers following cysteamine administration shows a disappearance of the SS14 immunoreactive fibers and cell bodies with no apparent change in the SS28(1-12) immunoreactive fibers and cell bodies. These data suggest that, in rat hypothalamus, selective release of SS14 and SS28(1-12) can occur. The results are discussed in relation to possible sites of storage and release of the somatostatin-related peptides from synaptic nerve terminals.

Animals↗

PubMed1983

Immunohistochemical evidence that brain enkephalins arise from a precursor similar to adrenal preproenkephalin.

We have examined rat brains by immunohistochemistry with an antiserum raised against BAM22P, a synthetic fragment of the adrenomedullary precursor of Leu- and Met-enkephalin, in order to determine whether preproenkephalin derived peptides are detectable in the central nervous system. BAM22P antiserum stained fibers, nerve endings and cell bodies in many areas of the brain, especially the striatum, the septum and the hypothalamus. Comparative topographical studies showed that neurons recognized by anti-BAM22P overlapped structures found immunoreactive for enkephalins in the present study. In addition, BAM22P antiserum stained cell bodies in cerebral cortex and in caudate putamen. The presence of BAM22P immunoreactivity in presumptive enkephalin neurons suggests that some brain enkephalins arise from a precursor identical or closely related to the preproenkephalin A found in the bovine adrenal medulla. This does not preclude the possibility of enkephalins also arising from other precursor molecules.

Adrenal Medulla↗

PubMed1983

Immunohistochemical distribution of pro-somatostatin-related peptides in cerebral cortex.

Mammalian brain contains 3 peptides related to the pro-somatostatin molecule: somatostatin-14 (SS14), the form originally identified from hypothalamic extracts, somatostatin-28 (SS28) and somatostatin 28 (1-12) (SS28 (1-12)). By using antibodies which selectively recognize one or more of these 3 somatostatin-related peptides, we have characterized their immunohistochemical distribution in neocortex. These somatostatin-related peptides have a specific laminar distribution in cortex and are differentially distributed such that SS28 is largely restricted to cell bodies, whereas SS28 (1-12) is preferentially localized in neuronal processes and terminals in a density which far exceeds that revealed by SS-14 immunoreactivity. These data suggest that there may be an intraneuronal transformation from a SS28-like peptide to a SS28 (1-12)-like peptide. In addition, the enriched distribution of nerve fibers containing the antigenic determinant, SS28 (1-12), strongly implies that somatostatin-related peptides constitute a major neurotransmitter system in neocortex. The morphological characteristics of this system are homologous with long projection pathways such as the cortico-cortical specific intrinsic systems as well as projections.

Animals↗

PubMed1983

Release of somatostatin-28(1-12) from rat hypothalamus in vitro.

Following the discovery of the growth hormone release-inhibiting factor somatostatin from extracts of ovine hypothalamus, an N-terminally extended somatostatin of 28 amino acids has been identified in mammalian tissue. The original peptide, somatostatin-14 (SS14), corresponds to the C-terminus of somatostatin-28 (SS28). Both SS28 and SS14 have biological activity, occur in several rat brain regions, are present in cell bodies and nerve terminals and can be released in vitro upon depolarization in a calcium-dependent manner. Further, high-affinity binding sites were described for SS14, which also bind SS28 (refs 20-23). Recently, a dodecapeptide which corresponds to the N-terminus of somatostatin-28, somatostatin-28(1-12), has been characterized in rat hypothalamus. Radioimmunological and immunohistochemical studies have indicated the presence of SS28(1-12)-like immunoreactivity in several cortical and subcortical regions of the rat brain. This peptide was found to be unevenly distributed with the highest concentration in the hypothalamus, and preferentially localized to dendritic and axonal processes and terminals. These observations suggest that SS28(1-12) may be a neurotransmitter. In this study, we describe a calcium-dependent release of a SS28(1-12)-like peptide from hypothalamic slices in vitro. This finding supports a neurotransmitter function for this peptide.

Animals↗

PubMed1983

Inhibitory effects of cysteamine on neuroendocrine function.

The action of cysteamine on anterior pituitary hormone secretion was studied in vivo using conscious, freely moving male rats and in vitro using anterior pituitary cells in monolayer culture. Administration of 500 micrograms cysteamine into the lateral cerebral ventricles of normal rats caused the complete inhibition of pulsatile GH secretion for a minimum of 6 h. This treatment also significantly decreased plasma concentrations of LH for at least 6 h in orchiectomized rat, TSH in short-term (0.5 month) thyroidectomized rats, and PRL in long-term (6 months) thyroidectomized rats. The in vivo stimulation of GH, LH, TSH and PRL with their respective releasing hormones 60 min after administration of cysteamine was not different from the response observed in rats pretreated with saline except for PRL where cysteamine pretreatment significantly inhibited the expected PRL increase. In vitro, 1 mM cysteamine decreased basal and TRH stimulated PRL release while not affecting basal or stimulated GH, LH, TSH and ACTH secretion. These data demonstrate the dramatic and wide-ranging effects of cysteamine on anterior pituitary hormone secretion. This action appears to be mediated through hypothalamic pathways for GH, LH and TSH and through a pituitary pathway for PRL.

Animals↗

PubMed1983

Effects of cysteamine on pro-somatostatin related peptides.

Intracerebroventricular (icv) injection of cysteamine to rats produced a marked depletion of somatostatin-14-like immunoreactivity (LI) in all rat brain regions examined. The somatostatin-28 (SS28)-LI and SS28(1-12)-LI were generally not altered by the cysteamine treatment. Following subcutaneous injection of the drug similar depletions of hypothalamic SS14-LI was observed with no change in SS28-LI nor SS28(1-12)-LI. In vitro cysteamine significantly increased the basal release of SS14-LI and markedly potentiated the evoked release of SS14-LI from hypothalamic slices. At 10 mM cysteamine, enhanced release of SS14-LI from hypothalamic slices was still observed despite a marked depletion of tissue content of SS14-LI.

Animals↗

PubMed1983

Immunocytochemical localization of prosomatostatin fragments in maturing and mature secretory granules of pancreatic and gastrointestinal D cells.

Pancreatic and gastrointestinal D cells were examined by immunocytochemistry using antisera against somatostatin-28 (SS28) and its NH2-terminal fragment SS28-(1-12), followed by the staphylococcal protein A-gold (pAg) complex. In pancreatic and gastric D cells incubated with antiserum against SS28-(1-12) the gold particles produced intense staining of the mature secretory granules but weaker staining of the immature granules associated with the Golgi area, whereas after SS28 antiserum treatment the particles accumulated selectively over the population of immature secretory granules. In intestinal D cells not only SS28-(1-12) but also SS28 antiserum produced an intense gold staining over the mature delta granules. These observations show that the relative amounts of immunoreactive sites related to SS28 and its cleavage product SS28-(1-12) in maturing and mature secretory granules are different in pancreatic, gastric, and intestinal D cells.

Humans↗

PubMed1983

Pituitary adenomas that caused Cushing's disease or Nelson's syndrome are not responsive to ovine corticotropin-releasing factor in vitro.

The response of pituitary adenomas obtained surgically from patients with Cushing's disease of Nelson's syndrome to synthetic ovine corticotropin-releasing factor (CRF), vasopressins, somatostatin-28, dexamethasone, 3-isobutylmethylxanthine or high [K+] was examined in vitro by measuring the amount of pro-opiomelanocortin (POMC)-derived peptides secreted into the culture medium. CRF did not stimulate the secretion of adrenocorticotropin-, beta-endorphin-, or gamma 3-melanotropin-like peptides from the pituitary adenomas at concentrations ranging from 1 x 10(-13) M to 1 x 10(-7) M whereas vasopressins, 3-isobutyrl-methylxanthine and high [K+] increased, while somatostatin-28 and dexamethasone suppressed, the secretion of these POMC-derived peptides. These findings suggest that either the pituitary ACTH-producing tumors have lost their receptors to CRF or their post-receptor mechanism to CRF is not functional.

Adenoma↗

PubMed1983

Pancreatic somatostatinoma: abundance of somatostatin-28(1-12)-like immunoreactivity in tumor and plasma.

UNLABELLED: In the present study we characterized and compared the relative amounts of the different molecular forms of somatostatin-14 like immunoreactivity (S-14 LI) and of somatostatin-28(1-12) like immunoreactivity (S-28(1-12) LI) in extracts of tumor and peripheral plasma of a patient with a pancreatic somatostatinoma. Tissue and plasma were chromatographed on Sephadex G-50 columns equilibrated with 6 M urea. Immunoreactivity in the eluting fractions was assayed with two separate, region specific RIAs using antibodies R149 (S-14 LI) and S309 (S-28(1-12)LI). RIA R149 recognizes the 6-8 and 14 regions of the S-14 sequence and detects S-14, S-28, and prosomatostatin, an approximately 14,000 mol wt precursor for the two peptides. RIA S309 recognizes the 2-11 segment of S-28 and reacts with S-28, S-28(1-12), and higher mol wt S-28(1-12) LI but not S-14. Total tumor S-14 LI was 190 pmol/mg protein and consisted of three peaks of immunoreactivity of apparent 14,000 mol wt (14K S-14 LI), 3,200 mol wt (3.2K corresponding to S-28) and 1,600 mol wt (1.6K corresponding to S-14). The three peaks comprised, respectively, 7%, 57%, and 36% of total S-14 LI. Total tumor S-28(1-12) LI was 594 pmol/mg protein and eluted as four major peaks of immunoreactivity as follows: peak I (mol wt 15,000, 10% of total S-28(1-12) LI); peak II (mol wt 8,000, 20% of S-28(1-12) LI), peak III (corresponding to S-28, 19% of S-28(1-12) LI); peak IV (corresponding to S-28(1-12), approximately 50% of total S-28(1-12) LI). Total plasma concentration of S-14 LI was 714 pM, being made up of the three peaks found in tumor but in the following relative amounts (14K S-14 LI, 22%; 3.2K, 29%; 1.6 K, 49%). Plasma S-28(1-12) LI was 4 times higher (2879 pM) than S-14 LI and contained immunoreactivity corresponding to each of the four peaks found in the tumor. CONCLUSIONS: 1) The tumor and plasma concentrations of S-28(1-12) LI were greater than that of S-14 LI. 2) Both tumor and plasma S-14 LI and S-28 LI were heterogeneous and comprised species corresponding not only to S-14 but also S-28, S-28(1-12), prosomatostatin, and other higher mol wt forms of S-28.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenoma, Islet Cell↗

PubMed1982

Immunohistochemical distribution of pro-somatostatin-related peptides in hippocampus.

By using immune sera which recognize one or more of the 3 peptides, somatostatin-14 (SS14), somatostatin-28 (SS28) and somatostatin-28(1-12) (SS28(1-12)) we have characterized their immunohistochemical distribution in the hippocampal formation. There exist at least two independent neuronal systems containing pro-somatostatin-related peptides: an intrinsic system of cells in the polymorphic layers which branch locally, and a dense terminal field in the molecular layer of the dentate gyrus that may constitute a portion of the entorhinal-dentate projection. In addition, SS28 is the dominant form present in cell bodies, whereas SS28(1-12) is preferentially localized in neuronal processes and terminals. We could not detect SS14 immunohistochemically.

Animals↗