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Biomedical subjects

S E Leeman

Publications and source records attributed to S E Leeman.

At least 91 records · Page 5Linked to original sources
PubMed1984

Substance P immunoreactivity released from rat spinal cord after kainic acid excitation of the ventral medulla oblongata: a correlation with increases in blood pressure.

Kainic acid injections into the ventral medulla oblongata of sodium pentobarbital-anesthetized rats caused an almost immediate increase in mean arterial pressure which reached hypertensive levels (greater than 200 mm Hg). Correlated with this rise in blood pressure was a two-fold increase in the amount of SP-like immunoreactivity (SPLI) in spinal cord superfusion samples. Baseline values obtained from a pool of two rats went from approximately 10 pg/10 min to approximately 20 pg/10 min. Rats pretreated with intracerebroventricular injections of the serotonin neurotoxin, 5,7-dihydroxytryptamine (5,7-DHT), were also given kainic acid injections into the ventral medulla. 5,7-DHT did not block pressure response to kainic acid nor the elevation of SPLI in spinal cord superfusates. However, baseline levels of SPLI were decreased by approximately 50% in 5,7-DHT-treated rats. These findings show that activation of the ventral medulla oblongata causes an elevation of SPLI in spinal cord superfusates concomitant with an elevation in mean arterial blood pressure. These responses appear to be independent of serotonergic systems.

5,7-Dihydroxytryptamine↗
PubMed1984

Neonatal estrogen affects preoptic/anterior hypothalamic LHRH differently in adult male and female rats.

When administered during the critical perinatal period, estrogen permanently modifies both male and female reproductive function. This study evaluated the influence of exogenous estrogen administered during this time on the hypothalamic LHRH content and on gonadotropin secretion in adult male and female rats. LHRH content in the preoptic-anterior hypothalamic (PO/AH) and midhypothalamic (MH) areas of neonatally estrogenized rats (1 mg on postpartum day 2) and of control male and female rats during the estrous cycle was determined by radioimmunoassay between 80 and 90 days of age. LHRH content was significantly greater in the PO/AH of neonatally estrogenized females than in estrogenized males whereas no differences in LHRH content of the PO/AH existed between control male and female rats. Neonatal administration of estrogen resulted in increased LHRH content in the MH of female rats and decreased LHRH content in male rats, as in the PO/AH; however, these differences were less marked in the MH and not statistically significant. The marked increase in serum LH concentration present in control cycling females in proestrus was abolished by neonatal estrogen treatment. Exposure to neonatal estrogen reduced FSH concentrations in males. The data are consistent with the concept that the hypothalamic-pituitary system is modified by estrogen circulating during the period of sexual differentiation. LHRH synthesis and release appear to be directly modified by neonatal estrogen. The effects of neonatal estrogen treatment in the PO/AH and MH areas may possibly involve two disparate types of alterations of developing LHRH neurons which modulate gonadotropin secretion in the adult rat.

Animals↗
PubMed1984

Stimulation of luteinizing hormone release after stereotaxic microinjection of neurotensin into the medial preoptic area of rats.

Neurons immunoreactive to luteinizing hormone-releasing hormone (LHRH) are localized to the medial preoptico-septal region of the rat hypothalamus, an area known to be essential for the regulation of the surge of luteinizing hormone (LH) that precedes ovulation. The effect of neurotensin (NT) on circulating LH levels following its discrete microinjection into this region was evaluated. Within 30 min after the stereotaxic microinjection of 40 ng of NT in 50 nl into the medial preoptic area (MPOA) just rostral to the organum vasculosum of the lamina terminalis in ovariectomized (OVX), anesthetized rats, plasma LH levels rose from 417 +/- 36 to 923 +/- 113 ng/ml (p less than 0.001). There was also a significant increase (p less than 0.001) in plasma LH levels following the microinjection of 50 ng of norepinephrine in 50 nl in this same site. Similarly, levels of LH increased from 48 +/- 7 to 110 +/- 19 ng/ml (p less than 0.004) following the microinjection of NT into the rostral MPOA on the morning of proestrus in anesthetized rats. In contrast, saline, substance P, leu-enkephalin and LHRH microinjected in the rostral MPOA of OVX rats had no significant effect on circulating levels of LH. The microinjection of NT 0.4 mm rostral or caudal to this area was also without effect. The presence of two binding sites with KdS of 4.2 and 29 nM and site concentrations of 0.11 and 0.68 pmol/mg in tissue homogenates of the rostral MPOA were also established. These data suggest a role for NT in the central regulation of LH release.

Animals↗
PubMed1984

Regulation by thyroid hormone of the concentration of substance P in the rat anterior pituitary.

The content of immunoreactive substance P (iSP) in the male rat anterior pituitary was measured after thyroidectomy and excess T4 administration. Baseline values for iSP content (mean +/- SE, 400 +/- 37 fmol/mg protein) increased progressively after thyroidectomy (4 days, 893 +/- 100; 6 days, 1321 +/- 242; 14 days, 1897 +/- 509). Administration of pharmacological doses of T4 (50 micrograms daily) for 2 and 14 days significantly decreased anterior pituitary iSP content (2 days, 196 +/- 30; 14 days 138 +/- 12). Thyroid status did not affect iSP content in the hypothalamus, caudate nucleus, amygdala, or brainstem. Partial chemical characterization of SP immunoreactivity in the anterior pituitary was obtained by gel permeation chromatography on Sephadex G-25, high pressure liquid chromatography, and the use of two antisera in RIAs, one directed against the amino-terminus and one directed against the carboxyl-terminus of the peptide. SP in the anterior pituitary was readily releasable in vitro by 44 mM potassium chloride in a calcium-dependent manner. The present study demonstrates that the concentration of iSP in the rat anterior pituitary is affected by the thyroid status of the animal and supports the probability that thyroid hormones participate in the regulation of the synthesis and/or release of iSP from the anterior pituitary.

Animals↗
PubMed1984

The effects of gonadal steroids on the content of substance P in the rat anterior pituitary.

The effects of gonadectomy and of the administration of gonadal steroids on the content of substance P in the anterior pituitary (AP-SP) were studied in adult rats. The effect of gonadal status on the AP-SP content of thyroidectomized (TX) rats was also studied. We have confirmed that the AP-SP content in adult males is higher than that in adult females. Ovariectomy (OVX) caused an increase in AP-SP content which was apparent 6 days after surgery. Estradiol (E2; 2 micrograms/rat daily) administered for 13 days beginning the day after OVX prevented the increase in AP-SP content induced by OVX. Orchiectomy of adult rats had no effect on AP-SP content 14 and 45 days after surgery. E2 administered to adult female rats for 13 days caused a reduction in the AP-SP content, whereas dihydrotestosterone (0.2 mg/rat daily for 13 days) caused an increase that was even more pronounced in TX rats. E2 administration to TX adult female rats caused a significant decrease in the AP-SP content both when treatment was begun on the day after surgery or 2 weeks later. Administration of T4 (1.5 and 25 micrograms/100 g BW daily for 7 days) to rats made hypothyroid by thyroidectomy 2 weeks earlier abolished the increase in AP-SP content seen in TX animals. Neither E2 nor dihydrotestosterone had an effect on the substance P content of any of the brain regions examined. The AP-SP content of pregnant or lactating rats was not different from that of age-matched controls. The content of substance P in the AP and median eminence did not vary significantly throughout the estrous cycle. The data indicate that AP-SP content is dependent on the gonadal status of the animal and that gonadal steroids interact with thyroid hormones in the regulation of substance P turnover in the AP.

Animals↗
PubMed1983

Somatostatin is increased in the basal ganglia in Huntington disease.

Huntington disease (HD) is an autosomal dominant hereditary disorder characterized by premature cell death, predominantly in the neostriatum. Decreased concentrations of several neurotransmitters and neuropeptides have been reported in the basal ganglia in Huntington disease. We now report that concentrations of radioimmunoassayable somatostatin are increased in extracts of the caudate (mean +/- standard error of the mean, ng/gm net weight; 247 +/- 24 versus 85 +/- 11), putamen (275 +/- 48 versus 74 +/- 11), external globus pallidus (100 +/- 10 versus 27 +/- 6), and internal globus pallidus (108 +/- 21 versus 21 +/- 8) in the disease. The concentrations of immunoreactive substance P measured in the same extracts were markedly reduced in caudate (mean +/- standard error of the mean, pmol/gm wet weight; 25 +/- 3 versus 109 +/- 20), putamen (28 +/- 7 versus 88 +/- 28), external globus pallidus (39 +/- 9 versus 196 +/- 62), and internal globus pallidus (60 +/- 17 versus 263 +/- 39), as well as in both subdivisions of the substantia nigra. Gel permeation chromatography and high-performance liquid chromatography showed radioimmunoassayable somatostatin to include peptides with physicochemical properties of the tetradecapeptide somatostatin and larger substances, including somatostatin-28-like material. A single peak of immunoreactive substance P corresponding to synthetic substance P was found by high performance liquid chromatography. These results suggest that immunoassayable somatostatin-containing neuronal elements in the neostriatum and globus pallidus in Huntington disease are affected differentially by the disease process from neurons that contain immunoreactive substance P.

Adolescent↗
PubMed1983

Clonal strains of rat medullary thyroid carcinoma cells that produce neurotensin and calcitonin. Functional and morphologic studies.

Strains of rat medullary thyroid carcinoma cells were established by two different techniques which were designed to ensure that clonal populations originated from single cells. The clonal strains (44-3C1 and 6-23C6) secreted both immunoreactive calcitonin (CT) and neurotensin (NT) and had growth characteristics that were similar to those of the parent cell lines. Both the 44-3C1 strain and the parent 44-2 cell line consistently produced more CT and NT than the 6-23C6 clonal strain or the parent 6-23 cell line. Peptide secretion in these clonal strains was stimulated by calcium and norepinephrine. Both clonal strains, similar to the parent lines, produced 4 to 12 times more NT than CT. Immunohistochemical studies showed that all cells in both clonal strains stained positively for CT. NT was also present in all cells from both strains with approximately 2% of the cells showing intense staining for this peptide. Ultrastructurally, the cells contained membrane-bound secretory granules which had a mean diameter of 95 nm. Secretory granules were relatively numerous in only 2% of the cells where they tended to be concentrated in cell processes. These studies demonstrate that single rat medullary thyroid carcinoma cells produce both CT and NT and that these strains are useful models for studies of NT and CT biosynthesis and for investigations of the regulation of secretion of two peptides from a single cell type.

Animals↗
PubMed1982

Synaptic vesicles containing substance P purified by chromatography on controlled pore glass.

Subsynaptosomal particles containing the peptide, substance P, detected by radioimmunoassay were prepared by osmotic lysis of rat brainstem synaptosomes and chromatographed on a calibrated column of controlled pore glass beads of nominal pore size 300 nm. Immunoreactive substance P migrated on this column with particles of apparent mean diameter 117 nm. This particulate, immunoreactive substance P was indistinguishable from synthetic substance P by gel filtration and reverse-phase high pressure liquid chromatography. The specific activity of substance P with respect to protein in peak fractions was 140 pmoles/mg protein, 75-fold higher than in the crude homogenate. Particulate substance P was resistant to attack by endogenous proteases in a crude preparation. Depolarization of synaptosomes with 75 muM veratridine prior to vesicle purification decreased the recovery of particulate substance P 37%. This depletion was dependent on external calcium ions and was blocked by 1 muM tetrodotoxin. The foregoing properties of particulate substance P are consistent with its identification with the large, intraterminal substance P-positive vesicles seen by immunocytochemistry and suggest that these vesicles are involved in the release of substance P.

Animals↗
PubMed1982

Localization of neurotensin immunoreactive nerve fibers in the guinea-pig heart: evidence derived by immunohistochemistry, radioimmunoassay and chromatography.

By the use of the peroxidase-antiperoxidase technique guinea-pig hearts were investigated for the occurrence of neurotensin immunoreactivity. Neurotensin immunoreactive nerve fibers were found in distinct localisations in all hearts studied. In addition, neurotensin immunoreactive fibers were present in the adventitia of the ascending aorta, the aortic arch and the pulmonary trunk. All segments of the coronary vasculature exhibited a dense network of neurotensin immunoreactive fibers. This innervation pattern was most pronounced in the arterial portions. Neurotensin immunoreactive fibers occurred also in close contact with atrial and ventricular muscle cells. A particularly dense innervation by neurotensin immunoreactive fibers was present in the sinu-atrial node and in the atrio-ventricular node. The fibers were associated intimately with blood vessels as well as with nodal cells. In addition, neurotensin immunoreactive fibers were found in intracardiac ganglia. The presence of neurotensin-like immunoreactive material in the guinea-pig heart was demonstrated also by radioimmunoassay. The results of immunohistochemistry and radioimmunoassay were correlated. High performance liquid chromatographic analysis indicated that 20-30% of the total immunoreactivity co-chromatographed with guinea-pig or synthetic neurotensin. Evaluation of consecutive sections revealed different innervation patterns of neurotensin and substance P immunoreactive fibers. The findings suggest a neurotransmitter and/or neuromodulator function of neurotensin in the regulation of coronary circulation, of cardiac impulse generation and conduction, of heart muscle contractility and of cardiac reflex mechanisms. It is speculated that neurotensin might represent the efferent and substance P the afferent part of a cardiac regulatory system.

Animals↗
PubMed1982

Light microscopic and ultrastructural localization of immunoreactive substance P in the dorsal horn of monkey spinal cord.

Light- and electron-microscopic localization of substance P in the monkey spinal cord was studied by the peroxidase anti-peroxidase technique with the particular aim of examining types of interactions made by substance P-positive boutons with other neuronal elements in the dorsal horn. By light-microscopy dense labeling for immunoreactive substance P was found in laminae I, II (outer zone) and V (lateral region), consistent with findings in other mammalian species. By electron-microscopy, substance P-positive staining was mostly in unmyelinated and in some thinly myelinated small diameter fibers. Substance P-positive terminals contained both large granular vesicles (80-120 nm diameter), which were filled with reaction product, and clear round vesicles (40-60 nm). Substance P-positive large granular vesicles were sometimes observed near presynaptic sites and in contact with dense projection there. Immunoreactive substance P boutons were small to large in size (1-4 micron), formed synapses with somata and large dendrites and were the central axons of synaptic glomeruli where they were in synaptic contact with numerous small dendrites and spines. Substance P-labeled axons frequently formed synapses with dorsal horn neurons which were also postsynaptic to other types of axons. Substance P-positive profiles participated in numerous puncta adhaerentia with unlabeled cell bodies, dendrites and axons. Only rarely, some suggestive evidence was obtained indicating that axons might synapse onto substance P-containing boutons. Biochemical analysis of monkey spinal cord tissue extracts, undertaken to characterize more precisely the immunoreactive substances, indicated that only substance P and its oxide derivative were detected with the antiserum used in the immunocytochemistry. These morphological findings show that substance P is contained within a class of axon terminals, many of which have been shown previously in the monkey to originate from the dorsal root. The results suggest that modulation of substance P primary afferents terminating in the outer dorsal laminae of the monkey spinal cord occurs in part via axonal inputs onto dorsal horn neurons postsynaptic to the primary afferent.

Animals↗
PubMed1982

Neurotensin stimulates exocytotic histamine secretion from rat mast cells and elevates plasma histamine levels.

1. Neurotensin stimulated histamine release and granule extrusion when applied to isolated rat peritoneal mast cells. 2. This secretory response was prevented by the removal of calcium or energy and was not accompanied by the release of lactic dehydrogenase. 3. The secretory response produced by neurotensin was prevented by prior treatment of mast cells with cromoglycate. 4. The intravenous injection of neurotensin into anaesthetized rats produced a rapid and significant increase in the level of blood histamine that was dependent upon the dose of neurotensin. 5. Treatment of rats with compound 48/80, 24 hr before neurotensin, abolished the elevation in blood histamine caused by neurotensin. The intravenous injection of cromoglycate 1-2 min before neurotensin greatly reduced the response to neurotensin. 6. The intradermal injection of neurotensin (0.03-30 p-mole) increased capillary permeability in rats pre-treated intravenously with Evans Blue. This response was abolished by the antihistamine, diphenhydramine. Increasing the dose of neurotensin to 300 p-mole partially overcame this inhibition by diphenhydramine. 7. Our results demonstrate that neurotensin can elicit an exocytotic secretory response from isolated rat peritoneal mast cells and elevate histamine levels in blood. It is suggested that some of neurotensin's physiological effects may be due to stimulation of mast cell secretion.

Animals↗
PubMed1982

Elevation of plasma neurotensinlike immunoreactivity after a meal. Characterization of the elevated components.

The detection of an elevation in neurotensinlike immunoreactivity in peripheral plasma for several hours after a meal has been confirmed and shown to be primarily due to the presence of aminoterminal fragments of neurotensin (NT) rather than to NT itself. We have developed a procedure to separate and characterize these N-terminal cross-reacting substances, and to estimate the contributions of these constitutents to plasma neurotensinlike immunoreactivity. Gel chromatography of pooled plasma extracts on Sephadex G-25 followed by reverse-phase high pressure liquid chromatography indicated that peptides coeluting with NT and its N-terminal partial sequences NT(1-8) and NT(1-11) were present in plasma. Comparison of plasmas collected before and 1 h after a defined meal, in five experiments, demonstrated no change in C-terminal immunoreactivity and an 8- to 10-fold rise in N-terminal immunoreactivity. Chromatographic analysis of pooled pre- and postmeal plasma in four experiments showed that essentially all of this elevation in neurotensinlike immunoreactivity measured with an N-terminal directed antiserum was due to increases in NT(1-8) and NT(1-11), while NT itself, measured using a C-terminal directed antiserum, did not increase appreciably in peripheral plasma 1 h after the meal. Generation of tritiated substances with the same elution times as NT(1-8) and NT(1-11) did occur after incubation of [(3)H]NT with whole blood in vitro, providing supporting evidence that these fragments are metabolites of NT. The marked elevation in the circulating levels of these fragments reflects that an increased secretion of NT occurred in response to the test meal. The secreted NT may have acted as a hormone before it was metabolized, or it may only have had a local (paracrine) effect.

Adult↗
PubMed1981

The regional distribution of somatostatin, substance P and neurotensin in human brain.

The regional distribution of somatostatin-, substance P- and neurotensin-like immunoreactivity was determined in 41 areas of 10 human brains. Each peptide is distributed widely in the human central nervous system and for each the pattern of distribution is unique. No significant relationship was found between peptide levels and patient age, interval between death and autopsy, and tissue storage time prior to assay. The regional distribution of these peptides is similar to that seen in several animal species and the pattern of this distribution is consistent with the idea that peptides function as neurotransmitters within the central nervous system. The problems of using human post-mortem material for peptide assay are discussed.

Adult↗