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A Sarrieau

Publications and source records attributed to A Sarrieau.

48 records · Page 3Linked to original sources

Corticosteroid receptors in rat hippocampal sections: effect of adrenalectomy and corticosterone replacement.

Rat brain sections, located at the hippocampal level, were used to study the effect of bilateral adrenalectomy, with or without corticosterone treatment, on the number and affinity of corticosteroid binding sites. Adrenalectomy induces an increase of corticosterone receptor binding sites whereas adrenalectomy followed by in vivo corticosterone treatment produces a 50% decrease of binding site number. Increases and decreases of binding site number were not associated with a significant modification of the affinity for corticosterone. The present data show that in vivo corticosterone modulates its own number of binding sites demonstrated by in vitro binding on brain sections, in a manner which is reminiscent of changes in cytosol receptors demonstrated by conventional biochemical methods. Thus, this in vitro method provides an alternative way to study the plasticity of hippocampal glucocorticoid receptors.

Adrenalectomy↗

Characterization and autoradiographic distribution of neurotensin binding sites in the human brain.

The characteristics and topographical distribution of monoiodo 125I-Tyr3-neurotensin (NT) binding sites in normal human brain tissue were studied on brain sections and by quantitative autoradiography. Sections at the level of the substantia nigra show a dissociation constant and maximal binding capacity of 4.8 +/- 0.8 nM and 70 +/- 7 fmol/mg protein, respectively. High density of 125I-NT binding sites were mainly found in dopaminergic (DA)-rich areas such as the substantia nigra, the ventral tegmental area, the striatum and the nucleus accumbens, further supporting an interaction between NT and DA neurons in human brain.

Aged↗

Role of phospholipids in the binding activity of vasoactive intestinal peptide receptors.

Phospholipase digestion of rat intestinal epithelial cell membranes was performed in order to study the influence of membrane phospholipids on the binding activity of VIP receptors. Phospholipases A2 and C strongly (ED50 congruent to 4 X 10(-2) and 4 X 10(-1) micrograms/ml, respectively) and rapidly reduced 125I-VIP binding to membranes whereas phospholipase D was ineffective. This suggests an important role of both hydrophobic and hydrophilic groups of phospholipids on VIP receptor binding activity.

Animals↗

[Radioautographic localization of neuropeptide receptors in the central nervous system].

The first step of any physiological effect of a neuropeptide (NP) is its recognition by specific receptor sites. The very organization of the central nervous system (CNS) does not permit a precise localization of these binding sites by conventional binding assays. The aim of the present paper is to describe in detail a recently developed in vitro methodology for the localization, visualization and quantitation of specific binding sites for various NP such as TRH, neurotensin and vasoactive intestinal peptide (VIP) in the rat CNS. The combination of this autoradiographic technique with radioimmunological measurements of NP, reveals that the endogenous distribution of THR, for example, in various brain regions, is not correlated with the presence of its binding sites. In vitro autoradiography may also be used to study the neurotransmitter/neuromodulatory role of NP in the CNS. This point will be illustrated by the effect of VIP on serotonin binding sites in both rat suprachiasmatic nucleus and hippocampal formation. Besides, the importance of the endocrine environment of the target tissue for NP action will also be discussed.

Animals↗

In vitro binding of tritiated glucocorticoids directly on unfixed rat brain sections.

We describe a new technique for measuring specific in vitro binding of tritiated adrenal steroids on unfixed cryostat brain sections. The specific binding of [3H]corticosterone represents about 70% of the initial binding. Kinetic studies show that specific binding for [3H]corticosterone reaches equilibrium after 15 min incubation at room temperature. Scatchard analysis of [3H]corticosterone in vitro binding gives a linear plot with an apparent dissociation constant (Kd) and a number of binding sites (Bmax) in the range of 10(-8) M and 100 fmol/mg protein, respectively. [3H]Dexamethasone binding under the same conditions gives a similar Kd and a Bmax of 55 fmol/mg protein. The order of potency for the relative binding affinity for [3H]corticosterone labeled sites is as follows: corticosterone greater than progesterone, dexamethasone, RU 38486 (a "pure" antiglucocorticoid), RU 26988 (a "pure" glucocorticoid), aldosterone greater than estradiol, testosterone. Anatomical studies reveal that sections at the level of the hippocampus bind more [3H]corticosterone and [3H]dexamethasone in vitro than more rostral sections taken at the level of the septum. Adrenalectomy increases the capacity of [3H]corticosterone to bind to these sites and perfusion of the brain to remove transcortin and other blood proteins does not modify [3H]corticosterone binding. We conclude that it is possible to measure in unfixed frozen brain sections glucocorticoid binding sites.

Adrenalectomy↗

In vitro biochemical characterization and autoradiographic distribution of 3H-thyrotropin-releasing hormone binding sites in rat brain sections.

In the present study, we describe the biochemical characteristics and the autoradiographic distribution of thyrotropin-releasing hormone (TRH) receptors in the rat central nervous system (CNS) after in vitro incubation of brain slices with 3H-TRH. Scatchard analysis showed that, in the range of concentrations tested (0.7-35 nM), 3H-TRH bound to a single-class of receptors with a dissociation constant of 6 nM and a number of binding sites of 20 fmol/mg protein. Increasing concentrations of unlabeled TRH produced a dose-dependent inhibition of 3H-TRH binding. The only analogue as potent as TRH to displace 3H-TRH binding was 3-Me-TRH, whereas 1-Me-TRH or TRH-free acid as well as pGlu-His, pGlu-Pro-NH2 or His-Pro-diketopiperazine were ineffective. Neither Luteinizing hormone-releasing hormone (LHRH), neurotensin, somatostatin, D-Ala-Met-enkephalin nor VIP showed any significant affinity for TRH binding sites. Autoradiograms obtained by apposition of LKB 3H-Ultrofilm showed that the highest concentrations of 3H-TRH binding sites were found in the ventral dentate gyrus of the hippocampal formation, the lateral amygdaloid nucleus, the nucleus accumbens, and the thalamic paraventricular nucleus. The biochemical characterization of 3H-TRH binding in brain sections is in good agreement with previous reports on membrane preparations and the autoradiographic localization of the binding sites provides anatomical support for the effects of TRH in the CNS.

Amygdala↗

Intestinal VIP receptors: differential effect of trypsin on the high and low affinity binding sites.

The effects of trypsin treatment on VIP binding to rat intestinal epithelial cell membranes were examined. The decrease in specific binding of [125I]VIP is dependent on the amount of trypsin used and digestion time. Specific binding decreases by 50% after 8 min with 20 micrograms/ml trypsin. Trypsin is active in the 1-100 micrograms/ml concentration range (ED50 approximately equal to 5 micrograms/ml). Non-specific binding is unaltered by the enzyme. The effect of trypsin is abolished by trypsin inhibitor. Scatchard analysis of VIP binding reveals two types of binding sites: sites I characterized by a high affinity, a low capacity and a high sensitivity to low trypsin levels (1-5 micrograms/ml); sites II characterized by a low affinity, a high capacity, resistant to low trypsin levels (1-5 micrograms/ml) but sensitive to a high trypsin level (20 micrograms/ml). Trypsin decreases the binding capacity by lowering the site number without altering their affinity. Sites not destroyed by trypsin retain their functional characteristics: KD, sensitivity to GTP and coupling with adenylate cyclase. It is concluded that sites I and II are proteins with different structures and/or differently localized in the membrane.

Animals↗

[Binding of tritiated corticosterone by brain sections of adrenalectomized rats].

In the present work we describe a new technique which permits us to study the specific binding of tritiated corticosterone in brain sections of adrenalectomized Rats. Under these conditions, the specific binding of the glucocorticoid represents 60 to 70% of the initial binding. The apparent dissociation constant and the number of binding sites, determined by Scatchard analysis, are in the range of 10(-8) M and 100 fmoles/mg of protein respectively.

Adrenalectomy↗

No aging effect on hippocampal type II glucocorticoid receptors in two inbred mouse strains.

We report here on the type II glucocorticoid receptor concentration in the hippocampal cytosol of two inbred strains of mice, C57BL/6J and DBA/2J, aged 2 and 24 months. Glucocorticoid receptors are usually considered as mediating cytotoxic effects of glucocorticoids on hippocampal neurons and modulating hippocampus-dependent behaviors. Either a decrease in glucocorticoid receptors with aging or no effect of aging have been reported previously. In order to test whether strain differences may explain these conflicting results, we have measured, using [3H]RU 28362, the hippocampal glucocorticoid receptor concentrations in two inbred strains of mice showing differential modifications of some hippocampus-dependent behaviors with aging. Our results show that there is neither a strain nor an age effect.

Aging↗

[125I]vasoactive intestinal polypeptide binding sites: quantitative autoradiographic distribution in the rat spinal cord.

The quantitative autoradiographic distribution of [125I]vasoactive intestinal polypeptide (VIP) receptor binding sites was investigated in the rat spinal cord. [125I]VIP binding sites are discretely distributed, with a rostro-caudal gradient, along the longitudinal length of the cord; highest densities of sites being observed in its lumbar and sacral segments. In transverse sections, highest levels of [125I]VIP sites are present in laminae I and II, around the central canal, and in the parasympathetic lateral horn of the sacral segment. Moderate densities are seen along the medial border of the dorsal horn and the sympathetic lateral horn of the thoracic cord. Low amounts of labeling are observed in most structures of the ventral horn while white matter areas are apparently devoid of specific [125I]VIP binding. Thus, the distribution of spinal [125I]VIP receptor sites correlates well with that of VIP-like immunoreactive materials and support possible roles for this peptide in sensory neurotransmission and in the control of autonomic functions.

Animals↗

Asymmetrical distribution of hippocampal mineralocorticoid receptors depends on lateralization in mice.

Previous experiments showed that the activation of the hypothalamic-pituitary-adrenal (HPA) axis during stress was associated with behavioral lateralization used as a marker of population heterogeneity in mice. Furthermore, brain asymmetries have been demonstrated in neurotransmitter metabolism and neuroendocrine modulation. As the hippocampus modulates the activity of the HPA axis in stress and basal conditions, we postulated that hippocampal corticoid receptors may be asymmetrically distributed and that asymmetry may differ according to behavioral lateralization of animals. In order to answer these questions, binding capacity of mineralocorticoid (MR) and glucocorticoid (GR) receptors was determined in right and left hippocampi of mice previously selected for paw preference. The results show that regardless of behavioral lateralization, there was a tendency for a right dominance in MR binding capacity in the hippocampus but interestingly, the percentage of right/ total MR binding capacity was inversely correlated with individual paw preference scores. The affinity of MRs did not depend on behavioral lateralization. GR binding capacity was similar in each hemisphere and no relationship was found between GR binding capacity and paw preference scores. These results suggest that hippocampal receptors for corticoids may play an important role in the asymmetrical brain control of immune reactivity.

Animals↗