Characterization of hypothalamic noradrenaline receptors in the supraoptic nucleus and periventricular region of the paraventricular nucleus of mice in vitro

Neuroendocrine regulatory peptide (NERP)-3, derived from the neurosecretory protein VGF (non-aconymic), is a new biologically active peptide identified through peptidomic analysis of the peptides secreted by an endocrine cell line. Using a specific antibody recognizing the C-terminal region of NERP-3, immunoreactive (ir)-NERP-3 was identified in acid extracts of rat brain and gut as a 30-residue NERP-3 with N-terminal pyroglutamylation. Assessed by radioimmunoassay, ir-NERP-3 was more abundant in the brain, including the posterior pituitary (PP), than in the gut. Immunohistochemistry demonstrated that ir-NERP-3 was significantly increased in the suprachiasmatic nucleus, the magnocellular division of the paraventricular nucleus, and the external layer of the median eminence, but not in the supraoptic nucleus, after dehydration. The immunoreactivity was, however, markedly decreased in all of these locations after chronic salt loading. Intracerebroventricular administration of NERP-3 in conscious rats induced Fos expression in a subset of arginine vasopressin (AVP)-containing neurons in the supraoptic nucleus and the magnocellular division of the paraventricular nucleus. On in vitro isolated rat PP preparations, NERP-3 caused a significant AVP release in a dose-related manner, suggesting that NERP-3 in the PP could be an autocrine activator of AVP release. Taken together, the present results suggest that NERP-3 in the hypothalamo-neurohypophyseal system may be involved in the regulation of body fluid balance.

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Release of oxytocin and vasopressin by magnocellular nuclei in vitro: specific facilitatory effect of oxytocin on its own release

Journal of Endocrinology ◽

10.1677/joe.0.1020063 ◽

1984 ◽

Vol 102 (1) ◽

pp. 63-NP ◽

Cited By ~ 186

Author(s):

F. Moos ◽

M. J. Freund-Mercier ◽

Y. Guerné ◽

J. M. Guerné ◽

M. E. Stoeckel ◽

...

Keyword(s):

Paraventricular Nucleus ◽

Supraoptic Nucleus ◽

Male Rats ◽

Facilitatory Effect ◽

Vasopressin Release ◽

Paraventricular Nuclei ◽

Magnocellular Nuclei ◽

Oxytocin Release ◽

Supraoptic Nuclei

ABSTRACT The release of endogenous oxytocin and vasopressin by rat paraventricular and supraoptic nuclei in vitro during a 10-min period, 30 min after beginning the incubation, was measured radioimmunologically. Mean basal hormone release per 10 min and per pair of nuclei was: 128·4 ± 12·4 (s.e.m.) pg vasopressin (n = 15) and 39·0 ± 3·0 pg oxytocin (n = 66) for supraoptic nuclei from male rats; 273·9 ± 42·6 pg vasopressin (n = 11) and 34·2 ± 3·5 pg oxytocin (n = 15) for supraoptic nuclei from lactating rats; 70·0 ± 8·6 pg vasopressin (n = 52) and 21·8 ± 1·3 pg oxytocin (n = 68) for paraventricular nuclei from male rats; 59·1 ± 8·6 pg vasopressin (n = 10) and 27·0 ± 4·6 pg oxytocin (n = 16) for paraventricular nuclei from lactating rats. In male and lactating rats, both nuclei contained and released more vasopressin than oxytocin. For oxytocin alone, the paraventricular nucleus of male rats contained and released significantly less hormone than the supraoptic nucleus. This difference was not apparent in lactating rats. For vasopressin alone, the paraventricular nucleus contained and released significantly less hormone than the supraoptic nucleus in both male and lactating rats. When the hormone released was calculated as a percentage of the total tissue content the release was about 0·9% for oxytocin from both nuclei in male and lactating rats and also for vasopressin in lactating rats, but was only about 0·5% for vasopressin from both nuclei in male rats. The influence of oxytocin and analogues of oxytocin (including one antagonist) upon the release of oxytocin and vasopressin was studied. Adding oxytocin to the incubation medium (0·4–4 nmol/l solution) induced a dose-dependent rise in oxytocin release from both nuclei of male or lactating rats. A 4 nmol/l solution of isotocin had a similar effect to a 0·4 nmol/l solution of oxytocin, but arginine-vasopressin never affected basal release of oxytocin. In no case was vasopressin release modified. An oxytocin antagonist (1 μmol/l solution) significantly reduced basal oxytocin release and blocked the stimulatory effect normally induced by exogenous oxytocin, as did gallopamil hydrochloride (D600, 10 μmol/l solution), a Ca2+ channel blocker, or incubation in a Ca2+-free medium. These findings are discussed in relation to the literature on the central effects of neurohypophysial peptides. It may be concluded that the regulatory role of endogenous oxytocin in the hypothalamus on the milk-ejection reflex could result from its local release in the extracellular spaces of magnocellular nuclei. J. Endocr. (1984) 102, 63–72

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In vitro characterization of neural stem cells: Functional response to the enteric neurotrophin GDNF and co-culture with human colonic smooth muscle

Gastroenterology ◽

10.1016/s0016-5085(01)80307-3 ◽

2001 ◽

Vol 120 (5) ◽

pp. A62-A62

Author(s):

M MICCI ◽

R LEARISH ◽

P PASRICHA

Keyword(s):

Stem Cells ◽

Smooth Muscle ◽

Neural Stem Cells ◽

Functional Response ◽

In Vitro Characterization

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Biological and physico-chemical characterization of ultra high diluted Hydrastis canadensis and in vitro studies on mammalian cells

Allgemeine Homöopathische Zeitung ◽

10.1055/s-0037-1601241 ◽

2017 ◽

Vol 262 (02) ◽

pp. 2-76

Author(s):

N Chandrakar ◽

MK Temgire ◽

SG Kane ◽

AK Suresh ◽

J Bellare

Keyword(s):

Mammalian Cells ◽

Chemical Characterization ◽

In Vitro Studies ◽

Hydrastis Canadensis ◽

Physico Chemical

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Assays for Phospholipid-Dependent Formation of Thrombin and Xa: A Potential Method for Quantifying Lupus Anticoagulant Activity

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646437 ◽

1991 ◽

Vol 66 (04) ◽

pp. 453-458 ◽

Cited By ~ 15

Author(s):

John T Brandt

Keyword(s):

Specific Activity ◽

Anticoagulant Activity ◽

Factor Xa ◽

Clinical Importance ◽

Potential Method ◽

Quantitative Expression ◽

Increased Risk ◽

Apparent Association

SummaryLupus anticoagulants (LAs) are antibodies which interfere with phospholipid-dependent procoagulant reactions. Their clinical importance is due to their apparent association with an increased risk of thrombo-embolic disease. To date there have been few assays for quantifying the specific activity of these antibodies in vitro and this has hampered attempts to purify and characterize these antibodies. Methods for determining phospholipid-dependent generation of thrombin and factor Xa are described. Isolated IgG fractions from 7 of 9 patients with LAs were found to reproducibly inhibit enzyme generation in these assay systems, permitting quantitative expression of inhibitor activity. Different patterns of inhibitory activity, based on the relative inhibition of thrombin and factor Xa generation, were found, further substantiating the known heterogeneity of these antibodies. These systems may prove helpful in further purification and characterization of LAs.

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Characterization of the Original Christmas Disease Mutation (Cysteine 206 → Serine): From Clinical Recognition to Molecular Pathogenesis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648381 ◽

1992 ◽

Vol 67 (01) ◽

pp. 063-065 ◽

Cited By ~ 6

Author(s):

Sherryl A M Taylor ◽

Jacalyn Duffin ◽

Cherie Cameron ◽

Jerome Teitel ◽

Bernadette Garvey ◽

...

Keyword(s):

Nucleotide Substitution ◽

Novel Mutation ◽

Factor Ix ◽

Causative Mutation ◽

Coding Region ◽

Body Of Knowledge ◽

Polymerase Chain ◽

Splice Junctions

SummaryChristmas disease was first reported as a distinct clinical entity in two manuscripts published in 1952 (1, 2). The eponym associated with this disorder, is the surname of the first patient examined in detail and reported by Biggs and colleagues in a paper describing the clinical and laboratory features of seven affected individuals (3). This patient has severe factor IX coagulant deficiency (less than 0.01 units/ml) and no detectable circulating factor IX antigen (less than 0.01 units/ml). Coding sequence and splice junctions of the factor IX gene from this patient have been amplified in vitro through the polymerase chain reaction (PCR). One nucleotide substitution was identified at nucleotide 30,070 where a guanine was replaced by a cytosine. This mutation alters the amino acid encoded at position 206 in the factor IX protein from cysteine to serine. The non conservative nature of this substitution, the absence of this change in more than 200 previously sequenced factor IX genes and the fact that the remainder of the coding region of this gene was normal, all provide strong circumstantial evidence in favour of this change being the causative mutation in this patient. The molecular characterization of this novel mutation in the index case of Christmas disease, contributes to the rapidly expanding body of knowledge pertaining to Christmas disease pathogenesis.

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THE RELATION BETWEEN THE HYPOTHALAMIC NEUROSECRETION AND THE CORTICOTROPHIN RELEASE IN EXPERIMENTAL CONDITIONS

Acta Endocrinologica ◽

10.1530/acta.0.xxxiv0008 ◽

1960 ◽

Vol XXXIV (I) ◽

pp. 8-18 ◽

Cited By ~ 6

Author(s):

E. Kivalo ◽

U. K. Rinne

Keyword(s):

Paraventricular Nucleus ◽

Median Eminence ◽

Supraoptic Nucleus ◽

Acute Stress ◽

Neurosecretory Material ◽

Experimental Conditions ◽

Hypothalamic Cells ◽

The One ◽

Hypophysial Portal ◽

Acth Release

ABSTRACT Acute stress, chronic stress plus hydration, cortisone treatment, cortisone treatment plus dehydration were used as methods of investigation and the relation between the neurosecretory activity of the hypothalamic supraoptic nucleus and paraventricular nucleus and the neurosecretory material around the hypophysial portal vessels of the median eminence on the one hand and the corticotrophin release on the other hand, has been studied in the rat. Whereas stress stimulates both the activity of the above mentioned cells of the hypothalamus and the ACTH release, stress plus hydration causes a depression of these hypothalamic cells but nevertheless causes a marked ACTH release. Cortisone inhibits the activity of the cells in the supraoptic nucleus and the paraventricular nucleus as well as the ACTH release whereas cortisone plus dehydration causes stimulation but inhibits the ACTH release. In some stress and cortisone treatment groups the variations of the neurosecretory material around the hypophysial portal vessels and of the ACTH release were found to show a correlation. It is concluded that the activity of the cells of the supraoptic nucleus and the paraventricular nucleus and the ACTH release do not seem to have any definite connection, whereas some observations indicate that the neurosecretory material in the region of the median eminence around the hypophysial portal vessels may have some significance in ACTH release.

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