Expression of estrogen receptor-beta protein and mRNA in the cerebellum of the rat

doi:10.1016/S0304-3940(00)01221-0

Neuroscience Letters

Volume 288, Issue 2, 14 July 2000, Pages 115-118

https://doi.org/10.1016/S0304-3940(00)01221-0 Get rights and content

Abstract

Estrogen has been shown to play a modulatory role in cerebellar neuronal signaling. Recent reports have also shown that estrogen receptor beta (ER-β) mRNA is expressed in cerebellum. The purpose of the present study was to identify and map ER-β protein expression, and to determine the identity of the major splice variants of ER-β mRNA in the cerebellum. Polyclonal antibodies raised against the NH₃- and COOH-termini of rat ER-β were used for immunohistochemistry. Mapping of ER-β immunoreactivity was compared with the distribution of ER-β mRNA using in situ hybridization. We also determined, using RT-PCR, whether the ER-β mRNA was variably spliced in cerebellum. Our results show that in all cases the distribution of ER-β protein was identical to the distribution of ER-β mRNA. Both Purkinje cells and scattered cells in the granule cell layer, perhaps golgi cells, robustly expressed ER-β. Reverse transcription-polymerase chain reaction analysis showed that three splice variants in addition to wild type ER-β are expressed in rat cerebellum. However, wild type ER-β was the predominant form. These observations provide anatomical evidence that neurons in the cerebellum express ER-β and thus may be targets of estrogen action.

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Acknowledgements

This work was supported by grants from the NSF (96-04723 to RJH) and NIAAA (F31-AA05461 to RHP).

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The brain has traditionally been considered to be a target site of peripheral steroid hormones. On the other hand, extensive studies over the past thirty years have demonstrated that the brain is a site of biosynthesis of several steroids. Such steroids synthesized de novo from cholesterol in the brain are called neurosteroids. To investigate the biosynthesis and biological actions of neurosteroids in the brain, data on the regio- and temporal-specific synthesis of neurosteroids are needed. In the mid 1990s, the Purkinje cell, an important cerebellar neuron, was discovered as a major cell producing neurosteroids in the brain of vertebrates. It was the first demonstration of de novo neuronal biosynthesis of neurosteroids in the brain. Subsequently, neuronal biosynthesis of neurosteroids and biological actions of neurosteroids have become clear by the follow-up studies using the Purkinje cell as an excellent cellular model. Progesterone and estradiol, which are known as sex steroid hormones, are actively synthesized de novo from cholesterol in the Purkinje cell during development, when cerebellar neuronal circuit formation occurs. Importantly, progesterone and estradiol synthesized in the Purkinje cell promote dendritic growth, spinogenesis and synaptogenesis via their cognate nuclear receptors in the Purkinje cell. Neurotrophic factors may mediate these neurosteroid actions. Futhermore, allopregnanolone (3α,5α-tetrahydroprogesterone), a progesterone metabolite, is also synthesized in the cerebellum and acts on the survival of Purkinje cells. On the other hand, at the beginning of 2010s, the pineal gland, an endocrine organ located close to the cerebellum, was discovered as an important site of the biosynthesis of neurosteroids. Allopregnanolone, a major pineal neurosteroid, acts on the Purkinje cell for the survival of Purkinje cells by suppressing the expression of caspase-3, a crucial mediator of apoptosis. I as a recipient of Kobayashi Award from the Japan Society for Comparative Endocrinology in 2016 summarize the discovery of cerebellar and pineal neurosteroids and their biological actions on the growth and survival of Purkinje cells during development.
17 beta-estradiol synthesis modulates cerebellar dependent motor memory formation in adult male rats
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Neurosteroid 17 beta-estradiol (E2) is a steroid synthesized de novo in the nervous system that might influence neuronal activity and behavior. Nevertheless, the impact of E2 on the functioning of those neural systems in which it is slightly synthesized is less questioned.
The vestibulo-ocular reflex (VOR) adaptation, may provide an ideal arena for investigating this issue. Indeed, E2 modulates cerebellar parallel fiber-Purkinje cell synaptic plasticity that underlies encoding of VOR adaptation. Moreover, aromatase expression in the cerebellum of adult rodents is maintained at very low levels and localized to Purkinje cells. The significance of age-related maintenance of low levels of aromatase expression in the cerebellum on behavior, however, has yet to be explored. Our aim in this study was to determine whether E2 synthesis exerts an effective and persistent modulation of VOR adaptation in adult male rats.
To answer this question, we investigated the acute effect of blocking E2 synthesis on gain increases and decreases in VOR adaptation using an oral dose (2.5 mg/kg) of the aromatase inhibitor Letrozole in peri-pubertal and post-pubertal male rats.
We found that Letrozole acutely impaired gain increases and decreases in VOR adaptation without altering basal ocular-motor performance and that these effects were similar in peri-pubertal and post-pubertal rats.
Thus, in adult male rats neurosteroid E2 effectively modulates VOR adaptation in both of the periods studied.
These findings imply that the adult cerebellum uses E2 synthesis for modulating motor memory formation and suggest that low and extremely localized E2 production may play a role in adaptive phenomena.
Endocrine disorders and the cerebellum: from neurodevelopmental injury to late-onset ataxia
2018, Handbook of Clinical Neurology
Citation Excerpt :
This can be prevented by estrogen treatment and postmenopausal women on estrogen treatment show an increase in the gray matter of the cerebellum (Boccardi et al., 2006; Ghidoni et al., 2006; Robertson et al., 2009). Improved detection methods revealed the presence of estrogen receptors in the cerebellum, explaining the estrogen responsiveness of the cerebellum (Price and Handa, 2000; Mitra et al., 2003). Estrogen affects cerebellar functions by enhancing Purkinje cell responsiveness to glutamate, thus augmenting glutamatergic neurotransmission (Smith et al., 1988, 1989; Andreescu et al., 2007).
Hormonal disorders are a source of cerebellar ataxia in both children and adults. Normal development of the cerebellum is critically dependent on thyroid hormone, which crosses both the blood–brain barrier and the blood–cerebrospinal fluid barrier thanks to specific transporters, including monocarboxylate transporter 8 and the organic anion-transporting polypeptide 1C1. In particular, growth and dendritic arborization of Purkinje neurons, synaptogenesis, and myelination are dependent on thyroid hormone. Disturbances of thyroid hormone may also impact on cerebellar ataxias of other origin, decompensating or aggravating the pre-existing ataxia manifesting with motor ataxia, oculomotor ataxia, and/or Schmahmann syndrome. Parathyroid disorders are associated with a genuine cerebellar syndrome, but symptoms may be subtle. The main conditions combining diabetes and cerebellar ataxia are Friedreich ataxia, ataxia associated with anti-GAD antibodies, autoimmune polyglandular syndromes, aceruloplasminemia, and cerebellar ataxia associated with hypogonadism (especially Holmes ataxia/Boucher–Neuhäuser syndrome). The general workup of cerebellar disorders should include the evaluation of hormonal status, including thyroid-stimulating hormone and free thyroxine levels, and hormonal replacement should be considered depending on the laboratory results. Cerebellar deficits may be reversible in some cases.
Thyroid hormone- and estrogen receptor interactions with natural ligands and endocrine disruptors in the cerebellum
2018, Frontiers in Neuroendocrinology
Although the effects of phytoestrogens on brain function is widely unknown, they are often regarded as “natural” and thus as harmless. However, the effects of phytoestrogens or environmental pollutants on brain function is underestimated. Estrogen (17beta-estradiol, E2) and thyroid hormones (THs) play pivotal roles in brain development. In the mature brain, these hormones regulate metabolism on cellular and organismal levels. Thus, E2 and THs do not only regulate the energy metabolism of the entire organism, but simultaneously also regulate important homeostatic parameters of neurons and glia in the CNS. It is, therefore, obvious that the mechanisms through which these hormones exert their effects are pleiotropic and include both intra- and intercellular actions. These hormonal mechanisms are versatile, and the experimental investigation of simultaneous hormone-induced mechanisms is technically challenging. In addition, the normal physiological settings of metabolic parameters depend on a plethora of interactions of the steroid hormones. In this review, we discuss conceptual and experimental aspects of the gonadal and thyroid hormones as they relate to in vitro models of the cerebellum.
Distribution of SNAP25, VAMP1 and VAMP2 in mature and developing deep cerebellar nuclei after estrogen administration
2014, Neuroscience
Citation Excerpt :
Developmental changes are dramatic in the postnatal cerebellum (Sakamoto et al., 2001, 2003; Sato et al., 2007), when cerebellar levels of E2 are much higher than in the plasma (Sakamoto et al., 2003). During postnatal period estrogen receptors are highly expressed (Price and Handa, 2000; Jakab et al., 2001) and extremely elevated levels of E2 are essential for the synaptic organization of cerebellar circuitry (Sakamoto et al., 2001, 2003; Sato et al., 2007). In line with this background, in the present study we also wanted to test whether E2 changed the pre- and postsynaptic protein distribution patterns and also affected synaptic density.
Synaptosomal-associated protein of 25 kDa (SNAP25), vesicle-associated membrane protein 1 (VAMP1) and 2 (VAMP2) are components of soluble N-ethylmaleimide-sensitive fusion attachment protein receptors (SNARE) complex which is involved in synaptic vesicle exocytosis, a fundamental step in neurotransmitter release.
SNARE expression in cerebellum correlates with specific neurotransmitter pathways underlying synaptic diversification and defined synaptic properties.
In this study we firstly characterized the distribution of SNAP25, VAMP1 and VAMP2 in the nerve terminals of a defined cerebellar region, the deep cerebellar nuclei (DCN), of adult and newborn rats. Then, given the pivotal role of estradiol (E2) in the synaptic organization of the cerebellar circuitry in early postnatal life, we examined whether administration of E2 in the newborn DCN affected synaptic density and changed the distribution of the presynaptic proteins SNAP25, VAMP1 and VAMP2, together with post synaptic density protein 95 (PSD95).
Results showed that: (1) distribution of SNAP25, VAMP1 and VAMP2 in adult DCN differs significantly from that found in newborn DCN; (2) administration of E2 in the newborn DCN affected synaptic density and also changed the distribution of the pre- and postsynaptic proteins.
The differential distribution of SNAP25, VAMP1 and VAMP2 in nerve terminals of adult and newborn rats may correlate with specific stages of neuronal phenotypic differentiation. The effects of E2 on SNAP25, VAMP1, VAMP2, PDS95 and synaptic density suggest that pre- and postsynaptic proteins are under estrogenic control during development and that synaptic maturation can also be related with the activity of this steroid.
Estrogen receptor beta: Tissue distribution and the still largely enigmatic physiological function
2014, Journal of Steroid Biochemistry and Molecular Biology
In 1996, the molecular biology of E2 had to be reevaluated: in an effort to identify novel nuclear receptors or unknown isoforms of existing receptors Kuiper and colleague described the expression of a novel subtype of the estrogen receptor (ER) in rat prostate and ovary. Upon this pioneering discovery the already known ER was renamed ERα while the newly described ER was termed ERβ. In this review an attempt is made to summarize the current knowledge regarding the expression and function of ERβ in selected reproductive and non-reproductive organs under physiological conditions. The data suggest that ERβ may be considered as a dominant-negative regulator of ERα modulating transcriptional responses to estrogens. The ratio of ER α vs. β. within a cell may determine the cell sensitivity to estrogens and its biological responses to the hormone. Conclusion: It is not the ligand, it is the multiplicity of receptors which determines the plethora of estrogen actions.
This article is part of a Special Issue entitled ‘Phytoestrogens’.

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¹: Present address: Metabolic Research Unit, University of California San Francisco, San Francisco, CA 94143, USA.

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Expression of estrogen receptor-beta protein and mRNA in the cerebellum of the rat

Abstract

Section snippets

Acknowledgements

Brain Res. Dev. Brain Res.

Anal. Biochem.

Mol. Brain Res.

Brain Res.

Brain Res.

Long-term testosterone or diesthylstilbesterol treatment affects gamma-amino butyric acid and central-type benzodiazepine receptors but not peripheral-type benzodiazepine receptors in the female rat brain

Neuroendocrinology

Cytoplasmic estrogen receptors in rat brain: immunocytochemical evidence using three antibodies with distinct epitopes

Endocrinology

Cloning of a novel estrogen receptor expressed in rat prostate and ovary

Proc. Natl. Acad. Sci. USA

Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors α and β

Endocrinology

Expression and neuropeptidergic characterization of estrogen receptors (ER-α and ER-β) throughout the rat brain: anatomical evidence of distinct roles of each subtype

J. Neurobiol.