Optimization of immunohistochemical detection of rat ESR2 proteins with well-validated monoclonal antibody PPZ0506

Hattori, Yujiro; Ishii, Hirotaka; Higo, Shimpei; Otsuka, Mai; Kanaya, Moeko; Matsumoto, Kunihiro; Ozawa, Mina; Ozawa, Hitoshi

doi:10.1016/j.mce.2020.111145

Cited by 11 publications

(28 citation statements)

References 41 publications

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“…Recently, Andersson et al [1] validated thirteen commercially available monoclonal anti-ESR2 antibodies and identified only one antibody with sufficient specificity for human ESR2 proteins (PPZ0506). Our previous studies [12,17] confirmed its specific cross-reactivity against rodent ESR2 proteins and applicability for immunohistochemical detection in rats. Expression analyses of ESR2/ Esr2 transcripts and ESR2 proteins revealed distinct expression profiles in humans and rats.…”

Section: Introductionsupporting

confidence: 66%

“…Specifically, ESR2 transcripts and ESR2 proteins were highly expressed in the adrenal gland, immune system tissues, ovary, and testis, and widely distributed throughout the other organs in human [1,17]. In contrast, Esr2/ESR2 were highly expressed in the ovary, prostate, and certain neuronal nuclei, but scarcely found in other organs of rats [12,17].…”

Section: Introductionmentioning

confidence: 95%

“…Based on an assumption of wide distribution of mouse Esr2 transcripts and ESR2 proteins, phenotypes of Esr2knockout mice have been evaluated in multiple mouse organs and tissues, such as adipose tissues [27], immune tissues [9,19], lung [28,33], mammary gland [4,10,40], prostate [16,40,42], and uterus [39,41]. However, with the exception of ovary, prostate, and certain neuronal nuclei, the corresponding rat organs scarcely express rat Esr2 transcripts and ESR2 proteins [12,17]. Given this difference from rats, histological analysis is needed to determine whether ESR2 proteins are expressed in a wide range of organs in mice, as assumed.…”

Section: Introductionmentioning

confidence: 99%

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Optimized Mouse-on-mouse Immunohistochemical Detection of Mouse ESR2 Proteins with PPZ0506 Monoclonal Antibody

Ozawa

Hattori

Higo

et al. 2022

Acta Histochem. Cytochem.

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Despite the physiological significance of ESR2, a lack of well-validated detection systems for ESR2 proteins has hindered progress in ESR2 research. Thus, recent identification of a specific anti-human ESR2 monoclonal antibody (PPZ0506) and its specific cross-reactivity against mouse and rat ESR2 proteins heightened momenta toward development of appropriate immunohistochemical detection systems for rodent ESR2 proteins. Building upon our previous optimization of ESR2 immunohistochemical detection in rats using PPZ0506, in this study, we further aimed to optimize mouse-on-mouse immunohistochemical detection using PPZ0506. Our assessment of several staining conditions using paraffin-embedded ovary sections revealed that intense heat-induced antigen retrieval, appropriate blocking, and appropriate antibody dilutions were necessary for optimization of mouse-on-mouse immunohistochemistry. Subsequently, we applied the optimized immunostaining method to determine expression profiles of mouse ESR2 proteins in peripheral tissues and brain subregions. Our analyses revealed more localized distribution of mouse ESR2 proteins than previously assumed. Moreover, comparison of these results with those obtained in humans and rats using PPZ0506 revealed interspecies differences in ESR2 expression. We expect that our optimized methodology for immunohistochemical staining of mouse ESR2 proteins will help researchers to solve multiple lines of controversial evidence concerning ESR2 expression.

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Section: Introductionsupporting

confidence: 66%

Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

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Optimized Mouse-on-mouse Immunohistochemical Detection of Mouse ESR2 Proteins with PPZ0506 Monoclonal Antibody

Ozawa

Hattori

Higo

et al. 2022

Acta Histochem. Cytochem.

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“…A recent study [ 28 ] could not detect any significant amounts of ERβ in the human brain using a validated antibody against human ERβ. In contrast, follow-up studies using the same antibody did detect ERβ in the mouse brain after optimizing antigen retrieval [ 30 ] or using frozen mouse brain sections [ 31 ]. In line with this, the expression of ERβ mRNA has previously been shown widely distributed in the rodent brain [ 32 ], which is similar to the expression pattern demonstrated by Merchenthaler and colleagues [ 23 ] using an in-house generated ERβ antibody.…”

Section: Estrogen Receptors In the Brainmentioning

confidence: 99%

“…The preclinical data that exist on ERβ as a neuroprotective factor is very interesting from a therapeutic point of view. Since ERβ is mainly expressed in the brain, intestine and the gonads [ 23 , 28 , 30 ], it may be a promising target avoiding the side effects of systemic ERα activation. BBB permeable ERβ selective ligands are already in clinical trials for alleviating menopausal mood symptoms, and it would be interesting to explore their clinical potential in preventing neurodegeneration.…”

Section: Estrogen Receptors As Clinical Targets In Neurodegenerationmentioning

confidence: 99%

Estrogen receptors and the aging brain

Maioli

Leander

Nilsson

et al. 2021

Essays in Biochemistry

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The female sex hormone estrogen has been ascribed potent neuroprotective properties. It signals by binding and activating estrogen receptors that, depending on receptor subtype and upstream or downstream effectors, can mediate gene transcription and rapid non-genomic actions. In this way, estrogen receptors in the brain participate in modulating neural differentiation, proliferation, neuroinflammation, cholesterol metabolism, synaptic plasticity, and behavior. Circulating sex hormones decrease in the course of aging, more rapidly at menopause in women, and slower in men. This review will discuss what this drop entails in terms of modulating neuroprotection and resilience in the aging brain downstream of spatiotemporal estrogen receptor alpha (ERα) and beta (ERβ) signaling, as well as in terms of the sex differences observed in Alzheimer’s disease (AD) and Parkinson’s disease (PD). In addition, controversies related to ER expression in the brain will be discussed. Understanding the spatiotemporal signaling of sex hormones in the brain can lead to more personalized prevention strategies or therapies combating neurodegenerative diseases.

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