Immunohistochemical evidence of the presence of estrogen and progesterone receptors in the same neurons of the guinea pig hypothalamus and preoptic area

doi:10.1016/0006-8993(89)91561-8

Brain Research

Volume 480, Issues 1–2, 20 February 1989, Pages 1-15

https://doi.org/10.1016/0006-8993(89)91561-8 Get rights and content

Abstract

Estrogen and progesterone interact in the regulation of various brain functions including mechanisms controlling gonadotropin secretion and female sexual behavior. Various methods have been used to map the regions where these hormones act and where specific receptors can be detected. However, it remains unknown if both steroids act on the same neuron or if there are neural populations responding to either one or the other of these hormones. To answer this question, we used various immunocytochemical procedures to detect estrogen and progesterone receptors (ER-IR and PR-IR) on the same histological section, taking advantage of the fact that anti-ER monoclonal antibodies were raised from rats whereas anti-PR monoclonal antibodies were raised from mice. Initial experiments showed that the number of cells displaying ER and PR immunoreactivity changed with hormonal treatment. Prolonged treatment of ovariectomized guinea pigs with high doses of estradiol benzoate (EB) (10 or 15 μg/day for 4 or 6 days) increased the number of PR-positive cells in the preoptic area and the hypothalamus. Inversely, with this estrogen priming regimen the number of ER-positive cells decreased, as did the immunostaining intensity in their nuclei. With a lower dose of estrogen administered for a shorter period (2-μg dose of EB for 2 days) the two receptor populations could be easily observed. Under these hormonal conditions, the sequential procedure revealed that the vast majority of cells containing ER-IR in the preoptic area (nucleus preopticus periventricularis, medialis) and the mediobasal hypothalamus (nucleus periventricularis, arcuatus, ventrolateral part of nucleus ventromedialis and premamillaris) showed PR immunoreactivity. This was true regardless of the order in which dual immunocytochemical staining was performed. There were, however, a few ER-positive/PR-negative cells in these regions. However, only ER-positive neurons were found in the n.interstitialis stria terminalis, the n.amygdaloideus medialis and the n.supraopticus. Thus, in guinea pigs receiving moderate doses of estrogen, all PR-positive cells contain ER-IR whereas there is a population of ER-positive cells which are devoid of PR-IR.

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The rise in circulating estradiol between diestrus and proestrus, or the treatment of ovariectomized female rats with estrogen, were shown to induce PR in hypothalamic neurons (McGinnis et al., 1981a; Moguilewsky and Raynaud, 1979a; Romano et al., 1989; Scott et al., 2002). Double-label immunocytochemistry revealed that virtually all PR-positive hypothalamic neurons contain ER-immunoreactivity (Blaustein and Turcotte, 1989; Warembourg et al., 1989). The induction of hypothalamic PR by estrogen was then shown to be required for the facilitation of the lordosis response by progesterone.
Progesterone is commonly considered as a female reproductive hormone and is well-known for its role in pregnancy. It is less well appreciated that progesterone and its metabolite allopregnanolone are also male hormones, as they are produced in both sexes by the adrenal glands. In addition, they are synthesized within the nervous system. Progesterone and allopregnanolone are associated with adaptation to stress, and increased production of progesterone within the brain may be part of the response of neural cells to injury. Progesterone receptors (PR) are widely distributed throughout the brain, but their study has been mainly limited to the hypothalamus and reproductive functions, and the extra-hypothalamic receptors have been neglected. This lack of information about brain functions of PR is unexpected, as the protective and trophic effects of progesterone are much investigated, and as the therapeutic potential of progesterone as a neuroprotective and promyelinating agent is currently being assessed in clinical trials. The little attention devoted to the brain functions of PR may relate to the widely accepted assumption that non-reproductive actions of progesterone may be mainly mediated by allopregnanolone, which does not bind to PR, but acts as a potent positive modulator of γ-aminobutyric acid type A (GABA_A) receptors. The aim of this review is to critically discuss effects of progesterone on the nervous system via PR, and of allopregnanolone via its modulation of GABA_A receptors, with main focus on the brain.
Role of estrogen receptor α and β in the induction of progesterone receptors in hypothalamic ventromedial neurons
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Citation Excerpt :
There is the general belief that nuclear estrogen receptors (ERs) mediate most of the actions of estrogens in the VMNvl. Neurons in the VMNvl express ERs of both subtypes (α and β) and PRs (Warembourg et al., 1989). Although the ERα largely predominates, some of the VMNvl neurons express the ERβ or co-express both receptor subtypes (Shughrue et al., 1998; Shughrue and Merchenthaler, 2001).
The estrogen induction of progesterone receptors (PRs) in the ventrolateral division of the hypothalamic ventromedial nucleus (VMNvl) is critical for the regulation of female sexual behavior. VMNvl neurons express PRs and both types of estrogen receptors (ERα and ERβ), and their sequential activation initiates the molecular mechanisms underlying sexual behavior. To assess the relative importance of each ER subtype in the induction of PRs, we have estimated the total number of PR-immunoreactive neurons and quantified the total amount of PR protein in the VMNvl of adult ovariectomized rats that were injected with either estradiol benzoate (EB) or the specific agonists of the ERα, propyl-pyrazole triol (PPT), and of the ERβ, diaryl-propionitrile (DPN), in different doses and schedules. The administration of EB and of PPT alone, but not of DPN alone, increased the total number of PR-immunoreactive neurons and PR protein levels. When the specific agonists were administered sequentially, the total number of PR-immunoreactive neurons also increased, particularly when PPT was administered before DPN. Conversely, the concomitant administration of PPT and DPN did not increase the number of PR-immunoreactive neurons.
The observation that PPT increases the number of PR-immunoreactive neurons and the levels of PR protein far less than EB shows that the estradiol induction of PRs in the VMNvl does not involve solely the activation of the ERα and suggests that it might also implicate the activation of membrane receptors. The present results also show that ERβ activation averts the action of ERα in the induction of PRs.

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Research reportImmunohistochemical evidence of the presence of estrogen and progesterone receptors in the same neurons of the guinea pig hypothalamus and preoptic area

Abstract

Physiol. Behav.

Brain Research

Neurochem. Int.

J. Steroid Biochem.

Brain Research

Horm. Behav.

J. Biol. Chem.

Horm. Behav.

Steroids

Physiol. Behav.

Brain Research

Brain Res. Bull.

J. Steroid. Biochem.

Horm. Behav.

Brain Research

Neurosci. Lett.

Brain Research

Horm. Behav.

Recent studies on the regulation of pituitary LH and RSH secretion

Biol. Reprod.

Modulation by continuous oestrogen treatment and by progesterone of oestrogen receptors in the rabbit uterus

J. Endocrinol.

Estrogen-induced and estrogn-facilitated female rat sexual behavior is not mediated by progestin receptors

Neuroendocrinology

Interconnectedness of steroid hormone-binding neurons: existence and implications

Localized behavioral effects of tritiated estradiol implants in the ventromedial hypothalamus of female rats

Endocrinology

Progesterone: its role in the central nervous system as a facilitator and inhibitor of sexual behavior and gonadotropin release

Ann. N.Y. Acad. Sci.

The sites of action of ovarian steroids in the regulation of LH secretion

Neuroendocrinology

Monoclonal antibodies to human estrogen receptor

Research report
Immunohistochemical evidence of the presence of estrogen and progesterone receptors in the same neurons of the guinea pig hypothalamus and preoptic area