Characterization of 3H-AVP binding sites in particulate preparations of rat brain

Af, Pearlmutter; Mg, Costantini; Loeser, Bonnie K.

doi:10.1016/0196-9781(83)90144-4

Cited by 72 publications

(18 citation statements)

References 20 publications

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“…Thus sites with high affinity to [JH]-AVP and low affinity to [3H]-OT were described for the brain stem [13], the punched central amygdala region [21], for whole brain [27] and a variety of brain regions [14,39,42]. The unmasking of low-affinity AVP binding sites in vitro after exposure to divalent cations such as Ni2* or with pH values above 7.8, as demonstrated by Pearlmutter et al [39] and Junig et al [27], could be the counterpart of the low-af finity AVP binding sites found associated in abundance with the dentate gyrus [4,5], which did not respond to estra diol in the present study either. The above-mentioned puta tive AVP and OT receptor systems are also distinctly differ ent in distribution and binding specificity from the binding sites for a relatively stable enzymatically generated biologi cally active intermediate of the neurohypophyseal hor mones [9].…”

Section: Discussionmentioning

confidence: 99%

Estradiol Modulates Density of Putative Oxytocin Receptors’ in Discrete Rat Brain Regions

Kloet¹,

Voorhuis²,

Boschma³

et al. 1986

Neuroendocrinology

202

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Previous studies have provided evidence for a discrete localization of two types of vasopressin (AVP)-labeled binding sites in the rat brain, i.e., regions labeled preferentially with AVP (putative AVP receptors) and regions labeled with AVP as well as oxytocin (OT). The latter binding sites are considered here as putative OT receptors. In the present study the effect of estradiol on the number of these putative receptor sites for OT and AVP was investigated in rat brain after daily subcutaneous administration of the steroid (10 µg/100 g body weight) to ovariectomized rats. Specific binding of [³H]-OT and [³H]-AVP was determined after in vitro incubation of frozen brain sections, autoradiography and quantitation of the images with computer-assisted densitometry. Estradiol increased the number of OT receptors at least 4-fold in the ventromedial nucleus of the hypothalamus, regions of the olfactory tubercle, the nucleus accumbens and occasionally in the organum vasculosum laminae terminalis. A smaller increase (two-fold) was noted in the central amygdala, while a tendency to a decrease in OT receptor number was noted in the olfactory nucleus and the ventral subiculum. Estradiol treatment permitted an estimation of binding constants of [³H]-OT-binding to a membrane fraction of microdissected ventromedial hypothalamic region {K_d: 1.3 nM, B_max: 19.9 fmol/mg protein). The number of putative AVP receptors in the lateral septum and in the nucleus tractus solitarii was not affected by estradiol. In conclusion, the OT receptor system is subject to modulation by estradiol in some discrete brain regions, but not in others. One of the principal sites of this interaction is located in the ventromedial nucleus, which is a hypothalamic region known to contain a high density of estradiol receptors and to respond to estrogens by changes in ultrastructure, protein synthesis, electrophysiology and sexual behavior. The results show that estradiol may sensitize these processes to OT and AVP by increasing the number of putative OT receptors.

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Section: Discussionmentioning

confidence: 99%

Estradiol Modulates Density of Putative Oxytocin Receptors’ in Discrete Rat Brain Regions

Kloet¹,

Voorhuis²,

Boschma³

et al. 1986

Neuroendocrinology

202

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“…The concentration that produces half-maximal stimulation of flank-marking behavior is 1 ng (in 100 nl saline) or approximately 5 PM. In several studies that examined AVP binding in rat brain, Scatchard analysis showed the existence of high-and low-affinity binding sites with dissociation equilibrium constants (Kd) of approximately 1 and 20 nM, respectively (Brinton et al, 1984;Lawrence et al, 1984;Pearlmutter et al, 1983). Without taking into consideration the diffusion and degradation of AVP that may occur following its injection into the brain, the concentration of peptide employed in these studies is only about 200-fold greater than these Kd.…”

Section: Inhibition Ofjlank Marking By Selective a Vp Antagonistsmentioning

confidence: 99%

A V1-like receptor mediates vasopressin-induced flank marking behavior in hamster hypothalamus

Albers¹,

Pollock²,

Simmons³

et al. 1986

J. Neurosci.

110

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A vasopressin-sensitive mechanism within the medial preoptic area-anterior hypothalamus (MPOA-AH) appears to be essential for expression of a complex behavior involved in olfactory communication in Golden hamsters called flank marking. The present study investigated whether the induction of flank marking by arginine-vasopressin (AVP) within the MPOA-AH is mediated by a receptor that is more similar to the vasopressor (Vl) or the antidiurectic (V2) AVP receptor. Adult male hamsters were anesthetized and implanted with a 26 gauge guide cannula stereotaxically aimed at the MPOA-AH and then microinjected with analogs of vasopressin, oxytocin, and selective Vl and V2 antagonists. Hamsters were tested for flank-marking behavior during a 5 or 10 min observation period following the injection of peptlde in a vehicle of 100 nl of saline. None of the 15 analogs of AVP and oxytocin produced more flank marking than the 50.8 + 16.2 and 76.8 f 4.4 (mean f SEW, II = 4) flank marks observed following injection of AVP at the 1 or 10 ng dose, respectively. The number of flank marks produced by each analog was found to be highly related to the pressor activity of that analog at both the 1 ng (p = +0.74, p < 0.01) and 10 ng (p = +0.82, p < 0.01) doses. In contrast, no statistically reliable relationship between flank marking and the antidiuretic activity of these analogs was found at either dose (1 ng: p = +O.O7,p > 0.05; 10 ng: p = +O.lO,p > 0.05). In a second series of studies, microinjection of 2 different Vl antagonists into the MPOA-AH significantly (p < 0.005) inhibited flank marking in response to AVP. However, microinjection of a V2 antagonist or saline did not inhibit flank marking. These data indicate that the receptors mediating the induction of 'flank marking within the MPOA-AH are more similiar to Vl (pressor) receptors than to V2 (antidiuretic) receptors. Scent marking is a major form of communication in mammals.Odors deposited in the environment can serve a variety of social functions from attracting mates to repelling competitors (Yahr, 1984). In the Golden hamster (Mesocricetus auratus), one form of scent marking, termed flank gland marking, is accomplished by rubbing a dorsal flank gland against vertical objects in the environment. Flank gland marking is commonly observed in both male and female hamsters and can be elicited by the odors of other hamsters and aggression during social encounters (Johnston, 1975).Several lines of evidence now indicate that neurons within the medial preoptic area-anterior hypothalamus (MPOA-AH) Received Dec. 2, 1985; revised Jan. 20, 1986; accepted Jan. 22, 1986. We wish to thank Professor Maurice Manning for providing several vasopressin analogs and for his comments on the manuscript. This work was supported by NIH Grants (to H.E.A.) and HD-18022 (to C.F.F. are critical for the expression of scent marking behavior. Lesions that destroy the MPOA-AH severely reduce scent-marking behavior in several mammalian species (Hart, 1974;Hart and Voith, 1978;Yahr, 1984). Implants of testoster...

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“…To confirm this postulate, it is essential to demonstrate the existence of specific AVP binding sites in the CNS. Our initial report of specific AVP binding sites in brain (21,22), which has been observed and characterized in several other laboratories (23)(24)(25)(26)(27), led us to undertake a more detailed investigation. Thus, we now report a comprehensive quantitative autoradiographic analysis of the regional distribution of putative AVP receptors in the rat brain and pituitary.…”

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confidence: 99%

Regional distribution of putative vasopressin receptors in rat brain and pituitary by quantitative autoradiography.

Brinton

Gee

Wamsley

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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Characterization of 3H-AVP binding sites in particulate preparations of rat brain

Cited by 72 publications

References 20 publications

Estradiol Modulates Density of Putative Oxytocin Receptors’ in Discrete Rat Brain Regions

Estradiol Modulates Density of Putative Oxytocin Receptors’ in Discrete Rat Brain Regions

A V1-like receptor mediates vasopressin-induced flank marking behavior in hamster hypothalamus

Regional distribution of putative vasopressin receptors in rat brain and pituitary by quantitative autoradiography.

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