E. Genazzani

The murine NPY-1 receptor gene. Structure and delineation of tissue-specific expression.

The murine gene for the NPY‐1 receptor subtype for neuropeptide Y was characterized by DNA sequencing and expression studies. It comprises three exons with a 6,400 bp 5′‐untranslated and a 80 bp internal intronic sequence. The 5′‐flanking region of this gene lacks TATA or CCAAT consensus sequences in the proximity to the multiple transcription initiation sites. A 1,300 bp genomic fragment of the 5′‐flanking region drives the expression of the lacZ reporter gene in NG108‐15 cells and primary cultured neurons but not in glial and human embryonic kidney cells. In addition, it contains consensus sequences for various transcription factors including cAMP‐ and glucocorticoid‐responsive elements.

Prolactin receptors on large granular lymphocytes: Dual regulation by cyclosporin A

Although evidence has been provided for a modulatory role of prolactin (PRL) on humoral and cell-mediated immune responses and PRL receptors have been found on T and B lymphocytes, no indications exist concerning the influence of PRL on natural killer (NK) activity nor has a structural basis for interaction been found on the NK effector cells (large granular lymphocytes, LGL). We show here that highly purified LGL express binding sites for PRL. The calculated receptor number was 660 per cell and the dissociation constant (Kd) was 3.0 X 10(-10) M. Since previous studies have reported that cyclosporin (CsA), an immunosuppressive agent used in organ transplant patients, affects the binding of PRL to T and B lymphocytes, but not to rabbit mammary gland cells, we investigated whether this compound could alter the binding of the hormone to LGL. At concentrations from 10(-7) to 10(-6), corresponding to the therapeutical range, CsA induced a complete inhibition of the PRL binding. By contrast, concentrations of CsA ranging from 10(-11) to 10(-9) increased the PRL binding to more than 100% of control levels. In addition to their antitumor role, LGL have been proposed to participate in graft versus host disease and in transplant rejection. The finding that CsA can differently affect PRL-receptor expression on LGL points to an involvement of CsA--PRL interactions in determining the output of these immune responses. In addition, these data strongly support the idea of a close relationship between the neuroendocrine and immune systems.

HISTAMINE‐SENSITIVE ADENYLATE CYCLASE IN HYPOTHALAMUS OF RAT BRAIN: H1 AND H2 RECEPTORS

Abstract— In in vitro experiments on rat hypothalamic homogenates the effects of biogenic amines such as histamine (HA), noradrenaline (NA), dopamine (DA), serotonin (5‐HT) and drugs such as isoprenaline (ISP), 2‐(2‐pyridyl)ethylamine (H2 stimulant—Hls), 4‐methyl‐histamine (H2 stimulant H2s), mepyramine (H1 antagonistp Hla), cimetidine (H2 antagonist—H2a) were tested on adenylate cyclase activity. HA possessed a powerful stimulating effect on hypothalamic adenylate cyclase activity, higher than that shown by the other substances.

[3H]N-methylscopolamine binding to muscarinic receptors in human peripheral blood lymphocytes: Characterization, localization on T-lymphocyte subsets and age-dependent changes

The properties of the binding of the muscarinic receptor ligands, [3H]quinuclidinyl benzilate ([3H]QNB) and [3H]N-methylscopolamine ([3H]NMS) in human mononuclear cells were compared. The binding of [3H]QNB showed a high, non-specific component and lack of saturability in both intact mononuclear cells and preparations of lysed mononuclear cell membranes. Conversely the specific binding of [3H]NMS had a high affinity and was saturable at concentrations greater than 30 nM in both intact and broken cells. Classical muscarinic receptor antagonists displaced specific binding of [3H]NMS binding according to the law of mass action, while displacement curves for pirenzepine and muscarinic agonists were very shallow (nH less than 1), suggesting the presence of more than one subtype of muscarinic receptor on mononuclear cell membranes. Binding studies with [3H]NMS to purified mononuclear cell subpopulations demonstrated that muscarinic binding sites were mainly localized on thymus-derived (T) lymphocytes and large granule lymphocytes. Moreover evidence is presented of an age-dependent increase of the density of muscarinic binding sites on T-lymphocytes. The results are discussed in terms of the usefulness of the binding of [3H]NMS in studying the physiological function of muscarinic receptors on human T-lymphocytes and their possible changes in patients with neurological diseases.

Effects of estradiol on the ontogenesis of striatal dopamine D1 and D2 receptor sites in male and female rats

Since estradiol (E2) either increases or reduces the number of dopamine receptors in the corpus striatum of adult rats, depending on the dose and length of administration, the sensitivity of the two receptor subpopulations (D1 and D2) to E2 during ontogenesis was investigated. Rats of both sexes received either 10 micrograms/kg E2 for 3 days or 50 micrograms/kg for 6 days, and were sacrificed at the age of 15, 21, 40 and 120 days. D1 receptors (identified by [3H]SCH 23390 binding) displayed no changes in density and affinity in function of age, sex or E2 dose, whereas the D2 receptors (identified by [3H]spiperone binding) fell after the lower dose in all groups, and the higher dose resulted in supersensitivity in males of all ages, but only in the 15-day-old females. These findings show that the effect of E2 is bivalent on D2 density only. The effect of its brief administration at a low dose is not sex-dependent, whereas at higher doses administered for longer periods it appears to involve mechanisms linked to sexual differentiation after birth.

Nerve growth factor modulates the expression of muscarinic cholinergic receptor messenger RNA in telencephalic neuronal cultures from newborn rat brain

The effect of nerve growth factor (NGF) on muscarinic receptor subtypes was investigated in a primary culture of telencephalic neurons prepared from neonatal rats. The treatment with 100 ng/ml of NGF significantly enhanced choline acetyltransferase (ChAT) activity and intracellular acetylcholine (ACh) content during cultivation. The same treatment induced an early transient increase of the number of muscarinic cholinergic receptors (mAChR), as measured by [3H]quinuclidinyl benzilate binding to cell homogenate, that was followed by a dramatic decrease of the receptor density from the 9th day of culture. Atropine completely prevented the decrease of the maximal number of muscarinic recognition sites induced by NGF. Prolonged exposure of telencephalic neurons to NGF also induced a significant reduction of the relative content of the messenger RNA (mRNA) encoding m1 and m3 receptors, while the m4 transcript was increased by the treatment. We suggest that the prolonged stimulation of cholinergic neurons by NGF induces a downregulation of m1 and m3 mAChR and their mRNAs on the postsynaptic site, while it increases the synthesis of the functionally distinct m4 receptor subtype, which might be presynaptically localized on cholinergic neurons. The transient increase of the receptor number that occurs at the first days of culture was not paralleled by changes in the relative content of mAChR mRNAs and might be associated with the trophic activity of NGF on cholinergic synapses during early development.

Tamoxifen counteracts estradiol induced effects on striatal and hypophyseal dopamine receptors

UNLABELLED We investigated the ability of Tamoxifen (TAM), an antiestrogen drug, to counteract the modifications induced by estrogens on dopamine (DA) receptors on striatum and on adenohypophysis of ovex female rats. Subacute treatment with 17 beta-estradiol (E2) at both low (0.1 micrograms/kg) and high (20 micrograms/kg) doses confirmed its ability to increase the number of striatal 3H-Spiperone (3H-SPI) binding sites in a dose dependent manner. By contrast in the pituitary, only high doses of estrogen were effective in reducing the number of DA receptors. We treated ovex female rats for 15 days with TAM alone or associated with E2, to see if these estrogenic effects could be suppressed by an antiestrogenic drug. TAM did not affect the number of striatal DA receptors, but significantly increased the adenohypophyseal DA binding sites, without varying their affinity. No changes were observed in pituitary and striatal DA receptor density, even when TAM was injected in association with estradiol. IN CONCLUSION TAM is able to counteract the effects estrogens have on DA receptors. However there is some evidence that it could influence the pituitary DA systems independently of its antiestrogenic activity.

Further study on the changes in the concentration of prolactin-binding sites in different organs of Xenopus laevis male and female, kept under dry conditions and then returned to water (their natural habitat)

The binding of 125I-labeled ovine prolactin (125I-oPRL) to membranes from the kidney and liver of Xenopus laevis male and female specimens (Experiment I) and from the kidney, epidermis, and liver of female specimens (Experiment II) (1) kept in an aquatic environment, (2) exposed for 2 weeks to dehydrating conditions, and (3) then placed back into their aquaria after exposure to dehydrating conditions (Experiment II) was studied. No significant sex differences in PRL binding to kidney, epidermis, and liver were found. A highly significant drop in PRL specific binding to the membranes from the kidney and epidermis is brought about in the specimens from both sexes exposed to dehydrating conditions. The results obtained by MgCl2 treatment in vitro of the membranes under study for an estimation of total PRL receptor concentrations seem to point to an actual decrease in PRL specific binding sites. The values of PRL specific binding to the membranes from the liver are not affected by dehydration of the animals (Experiment I and II) or their subsequent rehydration (Experiment II). In rehydrated females (Experiment II), PRL binding values were closely related to those recorded in females permanently maintained in water (controls).

Pituitary modulation of sulpiride action in the central nervous system of the rat

Abstract To verify whether the stimulation by sulpiride of hypothalamic adenylate cyclase was direct or mediated by release of pituitary hormones, the effect of sulpiride on female hypophysectomized rats was studied. In these animals sulpiride does not substantially modify hypothalamic adenylate cyclase, brain 3,4-dihydroxyphenylacetic acid (DOPAC) levels and serum prolactin concentrations. Chlorpromazine on the contrary inhibits hypothalamic adenylate cyclase activity and increases DOPAC levels both in intact and in hypophysectomized rats. As sulpiride hardly crosses the blood-brain barrier, its action on pituitary with prolactin release seems to be essential to start the neurochemical phenomena in the central nervous system.

Enhancement of pentobarbital narcosis by drugs competing on the serum protein binding

Abstract Prior subcutaneous administration of sulfaethylthiazole (SET), sulfamethazine (SMZ), sulfanilamide (SNA), salicylic acid (SA), doxycycline (DOXI), p -amino-salicylic acid (PAS) is able to enhance the intraperitoneal pentobarbital (PB) narcosis in mice. Such narcosis enhancement seems to be connected with competition between these drugs and PB for serum proteins, which results in an increase of the unbound PB and consequently a higher PB cerebral concentration. This higher concentration may be attributed to competition for serum protein binding, since the PB metabolism is unaffected by SET, SMZ, SNA, SA, DOXI and PAS.