Elsevier

Nutrition Research

Volume 94, October 2021, Pages 10-24
Nutrition Research

Estrogen regulates the expression of retinoic acid synthesis enzymes and binding proteins in mouse skin

https://doi.org/10.1016/j.nutres.2021.08.002Get rights and content

Abstract

Topical 17-beta-estradiol (E2) regulates the hair cycle, hair shaft differentiation, and sebum production. Vitamin A also regulates sebum production. Vitamin A metabolism proteins localized to the pilosebaceous unit (PSU; hair follicle and sebaceous gland); and were regulated by E2 in other tissues. This study tests the hypothesis that E2 also regulates vitamin A metabolism in the PSU. First, aromatase and estrogen receptors localized to similar sites as retinoid metabolism proteins during mid-anagen. Next, female and male wax stripped C57BL/6J mice were topically treated with E2, the estrogen receptor antagonist ICI 182,780 (ICI), letrozole, E2 plus letrozole, or vehicle control (acetone) during mid-anagen. E2 or one of its inhibitors regulated most of the vitamin A metabolism genes and proteins examined in a sex-dependent manner. Most components were higher in females and reduced with ICI in females. ICI reductions occurred in the premedulla, sebaceous gland, and epidermis. Reduced E2 also reduced RA receptors in the sebaceous gland and bulge in females. However, reduced E2 increased the number of retinal dehydrogenase 2 positive hair follicle associated dermal dendritic cells in males. These results suggest that estrogen regulates vitamin A metabolism in the skin. Interactions between E2 and vitamin A have implications in acne treatment, hair loss, and skin immunity.

Introduction

Estrogen regulates the hair cycle. There are 4 main stages of the hair cycle: anagen (growth), catagen (regression), telogen (rest) [1], and exogen (release) [2]. Topical 17-beta-estradiol (E2) applied during telogen inhibited hair growth by arresting hair follicles in telogen in CD-1 and C57BL/6 mice [3,4]. In contrast, the synthetic estrogen antagonist ICI 182,780 (ICI) increased anagen. Topical E2 applied during late anagen induced early catagen, but ICI did not reverse this [4]. Estrogen receptor alpha (ESR1) regulated both of these effects [4,5]; while estrogen receptor beta (ESR2) apposed the action of ESR1 on the anagen to catagen transition [4]. Further studies suggested that E2 increased transforming growth factor beta 2 to induce premature catagen and bone morphogenetic protein 4 to maintain telogen [6].

E2 is synthesized within the pilosebaceous unit (PSU; hair follicle and sebaceous gland) [[7], [8]-9]. Aromatase (cytochrome P450, family 19, subfamily a, polypeptide 1; CYP19A1) is the rate-limiting enzyme that converts testosterone/androstenedione to E2/estrone respectively [10]. Inhibiting CYP19A1 with letrozole significantly reduced E2 and estrone [7]. CYP19A1 localized to the PSU, but only one stage of the hair cycle was examined [11]. Estrone and E2 were also synthesized in anagen hair follicles independent of CYP19A1 [9]. Thus, E2 synthesized within the PSU may impact the PSU and regulate the hair cycle.

Vitamin A and synthetic retinoids also altered hair, the hair cycle, and sebaceous gland function in humans and rodents [12], [13], [14], [15], [16], [17]. In addition, alterations in retinoids led to alopecia [13,[18], [19], [20], [21], [22], [23], [24], [25], [26]]. Pharmacological doses of synthetic retinoids etretinate and acitretin arrested hair follicles in telogen leading to telogen effluvium in humans [25]. Dietary vitamin A (retinyl esters) also regulated the hair cycle in a dose dependent manner in C57BL/6J and C3H/HeJ mice and impacted both central centrifugal cicatricial alopecia and alopecia areata [21,24,27]. In addition, depletion of 2 different vitamin A metabolism proteins lead to accelerated anagen, suggesting that these proteins are important in maintaining telogen [22,28].

Retinoic acid (RA) is an active metabolite of vitamin A and is argued to be synthesized at or near the site of action. Precise levels of RA are regulated by a set of key enzymes and binding proteins. Synthesis of RA from retinol occurs by the action of 2 enzyme families [29]. Retinol dehydrogenases convert retinol to retinal, while retinal dehydrogenases (aldehyde dehydrogenase family 1, subfamily A1, A2, and A3; ALDH1A1-3) convert retinal to RA. Dehydrogenase/reductase SDR family member 9 (DHRS9) is a retinol dehydrogenase regulated by estrogen in the uterus [30]. Cellular RA binding protein type II (CRABP2) also associated with the ability of tissues to synthesize RA [31], [32], [33], [34]. CRABP2 transports RA to the nucleus, and delivers it to RA receptors alpha, and possibly beta, and gamma (RARA, RARB, RARG) [35,36]. RARs are part of the nuclear hormone superfamily of ligand activated transcription factors [37,38]. RA is degraded by Cytochrome P450, family 26, subfamily a, polypeptide 1 (CYP26A1) and subfamily b, polypeptide 1 (CYP26B1) [39]. DHRS9, ALDH1A1-3, CRABP2 and RARA, B, and G localized to the PSU in a hair cycle dependent manner [40].

Estrogen regulates RA synthesis proteins in a tissue specific manner. Estrogen increased ALDH1A2 and CRABP2 in the rat uterus, human endometrium, hippocampus, and rat choroid plexus [31,[41], [42], [43], [44], [45], [46]-47]. Estrogen also increased DHRS9 and RA synthesis in the rat uterus; as well as DHRS9 and ALDH1A1 in the rat choroid plexus [31,41,42,47]. In contrast, estrogen reduced ALDH1A1 in the rat uterus and prolactin cells of the rat anterior pituitary gland [31,41-43,48,49]. E2 also increased Cyp26b1 in the hippocampus [45,46]. ESR1 was likely responsible for most of this regulation. ESR1 specific agonists reduced Aldh1a1 in the pituitary [49]. In addition, ESR1 directly bound a classical estrogen response element (ERE) in the promoter of Cyp26b1 and the first intron of RARA [45,50]. Crabp2 promoter analysis found no classical EREs; but ESR1 bound a region of DNA containing an imperfect ERE, ERE half sites, and Sp1 sites [41]. The Aldh1a2 promoter also contains a classical ERE, but studies have not confirmed ER binding [51]. Thus, estrogen directly regulated several RA synthesis proteins via ESR1 in many tissues.

The purpose of this experiment is to determine if estrogen regulates the expression of RA synthesis proteins in the cycling hair follicle. The rationale for this experiment is that estrogen is produced in the PSU and regulates the hair cycle. RA synthesis components localized to the PSU and retinoids regulated the hair cycle. In addition, estrogen regulated RA synthesis components in other tissues. The hypothesis tested is that locally produced estrogen regulates RA synthesis in the cycling hair follicle in an autocrine or paracrine fashion by inducing expression of DHRS9, ALDH1A2, and CRABP2, but possibly not ALDH1A3. This study found that both exogenous and endogenous estrogen regulates several RA synthesis enzymes and binding proteins. However, more significant effects were seen with the E2 antagonist (ICI) than the CYP19A1 inhibiter letrozole. This report also identified locations where E2 and its inhibitors regulated RA synthesis enzymes and binding proteins.

Section snippets

Mice

The Jackson Laboratory Institutional Animal Care and Use Committee approved all mouse work (protocol number 05003) in compliance with the US National Research Council's Guide from the Care and Use of Laboratory Animals, the US Public Health Service's Policy on Humane Care, and Use of Laboratory Animals, and Guide for the Care and Use of Laboratory Animals. C57BL/6J wild-type (+/+) (hereafter referred to as B6) mice (JR# 664, The Jackson Laboratory, Bar Harbor, ME) were used. Mice were fed the

Estrogen impacted RA synthesis components in telogen, but with considerable variability

To test the hypothesis that locally produced estrogen regulates RA synthesis component expression a pilot study was performed. Male C57BL/6J (B6) mice (n = 3) with hair follicles in telogen were shaved and topically treated with 17-β-estradiol (E2), E2 receptor antagonist ICI 182-780 (ICI), epigallocatechin gallate (EGCG), EGCG plus E2, or vehicle (acetone). EGCG is a non-specific inhibitor of CYP19A1 [60,61]. The EGCG plus E2 group was included to see if E2 could rescue the loss of E2 produced

Discussion

The original hypothesis that locally produced estrogen regulates RA synthesis in the cycling hair follicle in an autocrine or paracrine fashion by inducing expression of DHRS9, ALDH1A2, and CRABP2, but possibly not ALDH1A3 was accepted in some locations, but rejected in other locations. Exogenous or endogenous estrogen regulated DHRS9, ALDH1A1, ALDH1A2, ALDH1A3, CRABP2, RARA, and RARB in a sex specific manner (Fig. 6). Female control mice expressed more DHRS9, ALDH1A2, ALDH1A3, and CRABP2:

Author contributions

HBE, DEO, LEK, and JPS conceptualized this study; HBE curated the data and administered the project; HBE, DEO, LEK, and JPS acquired funding, provided resources, and supervised the project; HBE, KAS, SO, DEO, LEK, and JPS developed methods; HBE, KAS, ANS, FJD, and JPS performed investigation; HBE, FJD, and WW performed statistical analysis; HBE and DEO validated antibodies, and SO validated QPCR. HBE wrote majority of the document. AHS wrote original draft of the estrogen receptor localization

Acknowledgment

We thank Latha Raju and the Skin Disease Research Center Molecular Biology and Mouse Pathology cores at Vanderbilt University Medical Center; and Wanyi Wang in the Texas Woman's University Center for Research Design and Analysis for their assistance. This work was supported in part by grants from the North American Hair Research Society and NIH (AR052009, AR041943, AR041943-14 5254, and AR052710). The authors declared no conflicts of interest.

References (90)

  • MYS Shih et al.

    Retinol esterification by DGAT1 is essential for retinoid homeostasis in murine skin

    J Biol Chem

    (2009)
  • MA Anzano et al.

    Growth, appetite, sequence of pathological signs and survival following the induction of rapid, synchronous vitamin A deficiency in the rat

    J Nutr

    (1979)
  • FJ Duncan et al.

    Endogenous retinoids in the pathogenesis of alopecia areata

    J Invest Dermatol

    (2013)
  • LZ Wu et al.

    Mice lacking the epidermal retinol dehydrogenases SDR16C5 and SDR16C6 display accelerated hair growth and enlarged meibomian glands

    J Biol Chem

    (2019)
  • JL. Napoli

    Physiological insights into all-trans-retinoic acid biosynthesis

    Biochim Biophys Acta Mol Cell Biol Lipids

    (2012)
  • JL. Napoli

    Interactions of retinoid binding proteins and enzymes in retinoid metabolism

    Biochim Biophys Acta

    (1999)
  • D Dong et al.

    Distinct roles for cellular retinoic acid-binding proteins I and II in regulating signaling by retinoic acid

    J Biol Chem

    (1999)
  • HB Everts et al.

    Immunolocalization of enzymes, binding proteins, and receptors sufficient for retinoic acid synthesis and signaling during the hair cycle

    J Invest Dermatol

    (2007)
  • XH Li et al.

    Cellular retinoic acid-binding protein II gene expression is directly induced by estrogen, but not retinoic acid, in rat uterus

    J Biol Chem

    (2003)
  • XS Wang et al.

    Analysis of mouse retinal dehydrogenase type 2 promoter and expression

    Genomics

    (2001)
  • S Muller-Rover et al.

    A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages

    J Invest Dermatol

    (2001)
  • JP Sundberg et al.

    Skin and adnexa

  • HB Everts et al.

    Immunolocalization of retinoic acid biosynthesis systems in selected sites in rat

    Exp Cell Res

    (2005)
  • HB Everts et al.

    Hair cycle-specific immunolocalization of retinoic acid synthesizing enzymes Aldh1a2 and Aldh1a3 indicate complex regulation

    J Invest Dermatol

    (2004)
  • MI Shamonki et al.

    Immunohistochemical expression of endometrial L-selectin ligand is higher in donor egg recipients with embryonic implantation

    Fertil Steril

    (2006)
  • K Satoh et al.

    Inhibition of aromatase activity by green tea extract catechins and their endocrinological effects of oral administration in rats

    Food Chem Toxicol

    (2002)
  • J Valladeau et al.

    Cutaneous dendritic cells

    Semin Immunol

    (2005)
  • F Conrad et al.

    Substantial sex-dependent differences in the response of human scalp hair follicles to estrogen stimulation in vitro advocate gender-tailored management of female versus male pattern balding

    J Invest Dermatol Sym Proc

    (2005)
  • JW Fluhr et al.

    Glycerol regulates stratum corneum hydration in sebaceous gland deficient (Asebia) mice

    J Invest Dermatol

    (2003)
  • RM Porter et al.

    Defolliculated (Dfl): a dominant mouse mutation leading to poor sebaceous gland differentiation and total elimination of pelage follicles

    J Invest Dermatol

    (2002)
  • Q Ju et al.

    Sex hormones and acne

    Clin Dermatol

    (2017)
  • S. Kovats

    Estrogen receptors regulate innate immune cells and signaling pathways

    Cell Immunol

    (2015)
  • MN Erkelens et al.

    Retinoic acid and immune homeostasis: a balancing act

    Trends Immunol

    (2017)
  • ID Iliev et al.

    Intestinal epithelial cells promote colitis-protective regulatory T-cell differentiation through dendritic cell conditioning

    Mucosal Immunol

    (2009)
  • M Guilliams et al.

    Skin-draining lymph nodes contain dermis-derived CD103(-) dendritic cells that constitutively produce retinoic acid and induce Foxp3(+) regulatory T cells

    Blood

    (2010)
  • EA Langan et al.

    Mind the (gender) gap: Does prolactin exert gender and/or site-specific effects on the human hair follicle?

    J Invest Dermatol

    (2010)
  • HB Chase et al.

    Critical stages of hair development and pigmentation in the mouse

    Physiol Zool

    (1951)
  • HS Oh et al.

    An estrogen receptor pathway regulates the telogen-anagen hair follicle transition and influences epidermal cell proliferation

    Proc Natl Acad Sci U S A

    (1996)
  • U Ohnemus et al.

    Hair cycle control by estrogens: catagen induction via estrogen receptor (ER)-alpha is checked by ER beta signaling

    Endocrinol

    (2005)
  • HM Hu et al.

    Estrogen leads to reversible hair cycle retardation through inducing premature catagen and maintaining telogen

    PLoS One

    (2012)
  • G Wehner et al.

    Estrone sulfate source of estrone and estradiol formation in isolated human hair roots: Identification of a pathway linked to hair growth phase and subject to site-, gender-, and age-related modulations

    J Clin Endocrinol Metab

    (2014)
  • G Nikolakis et al.

    Skin steroidogenesis in health and disease

    Rev Endocr Metab Disord

    (2016)
  • ME Sawaya et al.

    Immunohistochemical distribution of aromatase and 3B-hydroxysteroid dehydrogenase in human hair follicle and sebaceous gland

    J Cutan Pathol

    (1991)
  • CN Frazier et al.

    Cutaneous lesions associated with a deficiency in vitamin A in man

    Arch Intern Med

    (1931)
  • SB Wolbach et al.

    Tissue changes following deprivation of fat-soluble A vitamin

    J Exp Med

    (1925)
  • Cited by (0)

    1

    David E. Ong is deceased.

    View full text