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Sex steroid receptors in the ovarian follicles of the lizard Sceloporus torquatus

Published online by Cambridge University Press:  22 May 2023

Norma Berenice Cruz-Cano Open the ORCID record for Norma Berenice Cruz-Cano [Opens in a new window] ,
Uriel Ángel Sánchez-Rivera ,
Carmen Álvarez-Rodríguez ,
Mario Cárdenas-León  and
Martín Martínez-Torres
Norma Berenice Cruz-Cano*
Affiliation:
Posgrado en Ciencias Biológicas, Universidad Nacional Autónoma de México, Edificio D, 1° Piso, Circuito de Posgrados, Ciudad Universitaria, Coyoacán, Ciudad de México Laboratorio de Biología de la Reproducción, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Avenida de los Barrios s/n, Los Reyes Iztacala, Tlalnepantla Estado de México C.P. 54110, México
Uriel Ángel Sánchez-Rivera
Affiliation:
Laboratorio de Biología de la Reproducción, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Avenida de los Barrios s/n, Los Reyes Iztacala, Tlalnepantla Estado de México C.P. 54110, México
Carmen Álvarez-Rodríguez
Affiliation:
Laboratorio de Biología de la Reproducción, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Avenida de los Barrios s/n, Los Reyes Iztacala, Tlalnepantla Estado de México C.P. 54110, México
Mario Cárdenas-León
Affiliation:
Laboratorio de Hormonas Proteicas, Departamento de Biología de la Reproducción, Instituto Nacional de Ciencias Médicas y de la Nutrición Salvador Zubirán, Avenida Vasco de Quiroga No. 15, Colonia Belisario Domínguez Sección XVI, Tlalpan Ciudad de México, C.P. 14080, México
Martín Martínez-Torres*
Affiliation:
Laboratorio de Biología de la Reproducción, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Avenida de los Barrios s/n, Los Reyes Iztacala, Tlalnepantla Estado de México C.P. 54110, México
*
Corresponding authors: Norma Berenice Cruz-Cano; Email: [email protected]; Martín Martínez-Torres; Email: [email protected]
Corresponding authors: Norma Berenice Cruz-Cano; Email: [email protected]; Martín Martínez-Torres; Email: [email protected]
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Summary

Estradiol and progesterone have been recognized as important mediators of reproductive events in the female mainly via binding to their receptors. This study aimed to characterize the immunolocalization of the estrogen receptor alfa (ERα), estrogen receptor beta (ERβ) and progesterone receptor (PR) in the ovarian follicles of the lizard Sceloporus torquatus. The localization of steroid receptors has a spatio-temporal pattern that depends on the stage of follicular development. The immunostaining intensity of the three receptors was high in the pyriform cells and the cortex of the oocyte of previtellogenic follicles. During the vitellogenic phase, the granulosa and theca immunostaining was intense even with the modification of the follicular layer. In the preovulatory follicles, the receptors were found in yolk and additionally, ERα was also located in the theca. These observations suggest a role for sex steroids in regulating follicular development in lizards, like other vertebrates.

Keywords

Estrogen receptor alfaEstrogen receptor betaFollicular developmentLizardProgesterone receptor
Type
Research Article
Information
Zygote , Volume 31 , Issue 4 , August 2023 , pp. 386 - 392
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This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited.
Copyright
© The Author(s), 2023. Published by Cambridge University Press

Introduction

The hypothalamus–hypophysis–gonad axis orchestrates the seasonality of reproduction via intracrine, paracrine, and endocrine signalling (Norris, Reference Norris2018). In female lizards, as in other vertebrates, sex steroids are considered the major hormones involved in the reproductive events (development, gametogenesis, mating behaviour, and courtship among others; Edwards, Reference Edwards2005; Sánchez and Smitz, Reference Sánchez and Smitz2012; Ramírez-Pinilla et al., Reference Ramírez-Pinilla, de Pérez, Alvarado-Ramírez, Rheubert, Siegel and Trauth2015). Generally, estradiol (E2) and progesterone (P4) have a dynamic behaviour depending on the stage of the reproductive cycle in vertebrate females (Amey and Whittier, Reference Amey and Whittier2000; Rhen et al., Reference Rhen, Sakata, Zeller and Crews2000; Edwards and Jones, Reference Edwards and Jones2001; Wack et al., Reference Wack, Fox, Hellgren and Lovern2008; Kummrow et al., Reference Kummrow, Smith, Crawshaw and Mastromonaco2010).

Although genomic (receptor-mediated) and non-genomic signalling is recognized for sex steroid actions in other vertebrates, the second has been scantly studied in Squamata (Contrò et al., Reference Contrò, Basile and Proia2015; Yatsu et al., Reference Yatsu, Katsu, Kohno, Mizutani, Ogino, Ohta, Myburgh, van Wyk, Guillette, Miyagawa and Iguchi2016; Yaşar et al., Reference Yaşar, Ayaz, User, Güpür and Muyan2017). In genomic signal transduction, sex steroids exert their effects by activating nuclear receptors that bind to their promoter regions and modulate gene expression (Sato et al., Reference Sato, Miyagawa, Iguchi, Takei, Ando and Tsutsui2016; Garg et al., Reference Garg, Ng, Baig, Driggers and Segars2017). In general, sex steroid receptors are conformed by a N-terminal region that stimulates gene transcription (domain A/B), a DNA binding domain (C), a nuclear localization signal (D), and a ligand-binding region (E) (Medina-Laver et al., Reference Medina-Laver, Rodríguez-Varela, Salsano, Labarta and Domínguez2021). These receptors have shown high-functional plasticity due to their number of combinations in modulators, levels of activity, and responses (Jacobsen and Horwitz, Reference Jacobsen and Horwitz2012).

In female lizards, E2 promotes follicular development and the synthesis of vitellogenin (VTG) in the liver. This hormone increases with the onset of vitellogenesis and ovulation in various species (Jones, Reference Jones, Norris and Lopez2011; Ramírez-Pinilla et al., Reference Ramírez-Pinilla, de Pérez, Alvarado-Ramírez, Rheubert, Siegel and Trauth2015; Barbosa-Moyano et al., Reference Barbosa-Moyano, Rodríguez-Chaparro, Santos and Ramírez-Pinilla2020). The ovary is the principal source of E2 to exert its multiple effects via receptors in different tissues (Contrò et al., Reference Contrò, Basile and Proia2015). The estrogen receptor (ER) has two isoforms: the alfa (ERα) and beta (ERβ). These receptors were reported in many lizard species such as Bradypodion pumilum, Plestiodon finitimus, Gekko japinicus (Yatsu et al., Reference Yatsu, Katsu, Kohno, Mizutani, Ogino, Ohta, Myburgh, van Wyk, Guillette, Miyagawa and Iguchi2016), and Hemodactylus flaviviridis (Tripathy and Rai, Reference Tripathy and Rai2017). At least one receptor has been detected in Podarcis sicula (Verderame and Limatola, Reference Verderame and Limatola2010), Anolis carolinensis (Beck and Wade, Reference Beck and Wade2009), and Calotes versicolor (Inamdar et al., Reference Inamdar, Khodnapur, Nindi, Dasari and Seshagiri2015). In P. sicula, ERα is involved in vitellogenesis (Verderame and Limatola, Reference Verderame and Limatola2010), while in H. flaviviridis, it enhances aromatase activity via gonadotropin hormones (Tripathy and Rai, Reference Tripathy and Rai2017).

P4 is another hormone with pleiotropic effects in females (Cox, Reference Cox2020). This hormone also modulates vitellogenesis and ovulation (Crews, Reference Crews2005; Kabelik et al., Reference Kabelik, Weiss and Moore2008; Al-Amri et al., Reference Al-Amri, Mahmoud, Waring, Alkindi, Khan and Bakheit2012). However, its main roles are pregnancy maintenance and parturition in viviparous species due to its influence on uterine epithelium proliferation, uterine contractility inhibition, and downregulation of follicular development by modification of gonadotropin secretion and antiestrogenic capacity (Bonnet et al., Reference Bonnet, Naulleau, Bradshaw and Shine2001; Girling et al., Reference Girling, Jones and Swain2002; Martínez-Torres et al., Reference Martínez-Torres, Elena Hernández-Caballero, Alvarez-Rodriguez, Alba Luis-Díaz and Ortíz-López2003; Custodia-Lora et al., Reference Custodia-Lora, Novillo and Callard2004; Holmes and Cree, Reference Holmes and Cree2006; Adams et al., Reference Adams, Biazik, Stewart, Murphy and Thompson2007). The principal source of P4 is the corpus luteum (Hosie et al., 2003; Martínez-Torres et al., Reference Martínez-Torres, Elena Hernández-Caballero, Alvarez-Rodriguez, Alba Luis-Díaz and Ortíz-López2003, 2010, 2014). This progestogen shows low concentrations during follicular development, a peak before ovulation and the highest concentrations near oviposition or during embryo development (Jones, Reference Jones, Norris and Lopez2011; Ramírez-Pinilla et al., Reference Ramírez-Pinilla, de Pérez, Alvarado-Ramírez, Rheubert, Siegel and Trauth2015; Barbosa-Moyano et al., Reference Barbosa-Moyano, Rodríguez-Chaparro, Santos and Ramírez-Pinilla2020).

Two forms of progesterone receptor (PRA and PRB) have been reported and are associated with the stage of the reproductive cycle in reptiles (Custodia-Lora and Callard, Reference Custodia-Lora and Callard2002; Biazik et al., Reference Biazik, Parker, Murphy and Thompson2012; Hammouche et al., Reference Hammouche, Remana and Exbrayat2012; Motta et al., Reference Motta, Tammaro, Di Lorenzo, Panzuto, Verderame, Migliaccio and Simoniello2020). The difference between PRA and PRB resides in an extra-domain with activating function (AF) in the PRB (Garg et al., Reference Garg, Ng, Baig, Driggers and Segars2017). In the lizard Uromastix acanthinura, PRA and PRB are positively influenced by E2, which denotes heterogeneity in responses among species (Hammouche et al., Reference Hammouche, Remana and Exbrayat2012). In the turtle Chrysemys picta, RPA expression inhibits vitellogenin synthesis (Duggan and Callard, Reference Duggan and Callard2003), whereas RPB promotes it (Giannoukos and Callard, Reference Giannoukos and Callard1995). In Squamata, the diminution of uterine contractions may be regulated via RPA by limiting the activity of proteins involved in cellular adhesion (Custodia-Lora and Callard, Reference Custodia-Lora and Callard2002; Lovern, Reference Lovern, Norris and Lopez2011; Hoss et al., Reference Hoss, Garcia, Earley and Clark2014; Blackburn, Reference Blackburn and Skinner2018; Refsnider et al., Reference Refsnider, Clifton and Vazquez2019).

Recent research on lizard reproductive biology has evaluated changes in sex steroid concentration and the molecular components involved in these processes (Motta et al., Reference Motta, Tammaro, de Stasio, Borrelli and Filosa2004, Reference Motta, Tammaro, Di Lorenzo, Panzuto, Verderame, Migliaccio and Simoniello2020; Tammaro et al., Reference Tammaro, Simoniello, Filosa and Motta2008; Tripathy and Rai, Reference Tripathy and Rai2017). However, most of the studies involve oviparous species. Here we propose the spiny collared lizard (Sceloporus torquatus), a viviparous lizard with distribution in the central region of México (Guillette Jr. and Méndez-de la Cruz, Reference Guillette and la Cruz1993), to provide more information on the reproductive biology of lizards. The females begin vitellogenesis in summer, ovulate in autumn, are pregnant until the following spring, and give birth in April–June (Guillette and Méndez-de la Cruz, Reference Guillette and la Cruz1993; Ortiz et al., Reference Ortiz, de Oca, Ugarte, de Oca and Ugarte2001; Cruz-Cano et al., Reference Cruz-Cano, Sánchez-Rivera, Álvarez-Rodríguez, Dávila-Govantes, Cárdenas-León and Martínez-Torres2021). This work aimed to characterize the presence of estrogen receptors (α and β) and PR in the ovarian follicles (previtellogenic, vitellogenic and preovulatory) of the lizard S. torquatus and relate them to the concentration of these sex steroids.

Materials and methods

Animals

Fifteen sexually mature females (snout–vent length > 70 mm) of S. torquatus were collected in the Sierra de Guadalupe State Park (19°37′N, 99°12′W), State of México. The collecting scientific license was granted by the Secretaría del Medio Ambiente y Recursos Naturales (SEMARNAT; SGPA/DGVS/00579/17 and SGPA/DGVS/02921/19). The collection was carried out during follicular development (June to November).

Sample collection

Immediately after the capture, we collected a 100-µl blood sample by cardiac puncture with a heparinized syringe to measure sex steroid concentrations. The samples were taken to the Reproductive Biology Laboratory at 4ºC and were centrifuged to store the plasma until hormonal quantification. We anaesthetized females and performed a paramedian celiotomy to extract the ovarian follicles (Cruz-Cano et al., Reference Cruz-Cano, Sánchez-Rivera, Álvarez-Rodríguez, Dávila-Govantes, Cárdenas-León and Martínez-Torres2021). Then, the tissues were fixed with Bouin solution, and processed for routine histological technique and paraffin embedded.

As soon as the lizards fully recovered, we returned them to the site where they were collected.

E2 and P4 quantification

Hormone quantification was performed according to Cruz-Cano et al. (Reference Cruz-Cano, Sánchez-Rivera, Álvarez-Rodríguez, Dávila-Govantes, Cárdenas-León and Martínez-Torres2021). We used ASCENT software to obtain the values of the sex steroids. E2 was measured using a commercial immunoenzyme assay kit (Abcam ab108667, USA). The sensitivity was 8.68 pg/ml, according to the manufacturer’s protocol. The inter-assay and intra-assay coefficients of variation were 5.9% and 7.2%, respectively. P4 was quantified with a commercial radioimmunoassay kit (PROGEST-RIA, CISBIO). We used I125-labelled P4 as a reactive tracer. The sensitivity of the assay was 0.05 ng/ml. The inter-assay and intra-assay coefficients of variation were 4.6% and 7.5%, respectively.

Immunohistochemistry (IHC)

We obtained 5-µm sections from follicles of at least five lizards per stage (previtellogenesis, vitellogenesis, and preovulatory). The slides were deparaffinized in xylol, rehydrated through graded alcohols, and rinsed with distilled water. Immunohistochemistry was performed using the Vectastain Universal Quick Kit PK-8800 (Vector Laboratories Inc., Burlingame, CA, USA). Antigen retrieval was carried out with 0.1 M citrate buffer (pH 6.0) using heat treatment in a water bath set at 100ºC for 20 min. Quenching of endogenous peroxidase was made with 0.3% H2O2 for 30 min and washed in phosphate-buffered saline (PBS) 0.1 M (pH 7.4). Background-staining was blocked with preincubation with a solution containing horse serum (1 drop) and bovine serum albumin (BSA) 5% in 2 ml of PBS for 30 min and rinsed with PBS. Sections were incubated overnight in a humidified chamber at room temperature with the polyclonal primary antibodies (rabbit anti-ERα, Thermo Fisher PA5-16440; anti-Erβ, Thermo Fisher 51-7700; or anti-PR, Thermo Fisher PA5-94983, diluted 1:100). We incubated tissues with a biotinylated Pan-Specific Universal antibody for 15 min and washed them twice with PBS. Then, the slides were incubated with streptavidin/peroxidase complex for 10 min. We used 0.03%, diaminobenzidine tetrahydrochloride (DAB; Vector Laboratories) in PBS and 0.1% H2O2 to reveal the peroxidase activity. The sections were mounted in ClearVue (Thermo Fisher). The specificity and effectiveness of the IHC were assessed with two controls, a negative by omitting the primary antibody and a positive with mouse ovaries.

Statistical analysis

The values of E2 and P4 were assessed for normality with the Shapiro–Wilk test on raw data. We checked the homogeneity of the variances with Levene’s test. To determine the changes in the sex steroid concentrations among stages, we performed an analysis of variance (ANOVA) followed by a Holms–Sidak multiple comparison method. All the statistical analyses were performed with the significance level α = 0.05 using the SPSS program for macOS (Version 22).

Results

Ovarian follicles morphology

The morphology of the ovarian follicles presented changes associated with the stage of the reproductive cycle. The granulosa layer in the previtellogenic phase was composed of small, intermediate, and pyriform cells (Figure 1A). When the vitellogenesis began, the regression of the pyriform and intermediate cells reduced the height of the granulosa layer (Figure 1B). When the follicles reached the preovulatory stage, the small cells flattened, and the zona pellucida thickened (Figure 1C).

Figure 1. Morphology of ovarian follicles in the lizard S. torquatus. Previtellogenesis: (A) The follicles have a homogeneous ooplasm (o) with a cortex in the oocyte periphery (c); the granulosa is a multilayered structure with polymorphic cells small (s), intermediate (i) and pyriform (p). The zona pellucida (z) and a fibrous theca (t) with blood vessels (v) are clearly distinguishable. Vitellogenesis: (B) The oocyte contains yolk granules (y), and the granulosa is comprised by a monolayer of small cells (s), the zona pellucida (z) is thicker, the theca (t) has blood vessels (v) and mast cells (mc), and the small cells (s) begin to flatten. Preovulatory: (C) The zona pellucida (z) decreases in thickness and the small cells (s) are flattened in a single layer, also there is a decrease in the blood vessels (v). (A–C Masson’s Trichrome stained.)

Sex steroid concentrations

Sex steroid concentrations changed depending on the stage of follicular development (Table 1). E2 concentrations were low in the previtellogenic (254.61 ± 26.88 pg/ml) and vitellogenic stage (241.84 ± 40.71) and significantly higher in the preovulatory phase (625.95 ± 54.80) (ANOVA F(2,12) = 26.522, P < 0.001). For P4, there were no statistical differences among the stages studied (P > 0.05 in all cases) and ranged from 1464 to 1790 pg/ml.

Table 1. Plasma sex steroid concentrations in different phases of the follicular development in the lizard S. torquatus

* P < 0.01 vs previtellogenic and vitellogenic phase.

Sex steroid receptors

The previtellogenic follicles from June to September and November showed an intense mark of ERα in pyriform cells (Figure 2A) and granular structures in the cytoplasm of pyriform cells and the cortex (Figure 2B). However, these follicles showed signs of atresia, such as thickening of the theca, abundant small cells, and absence of a clearly defined zona pellucida.

Figure 2. Immunolocalization of ERα in atretic previtellogenic ovarian follicles. Previtellogenesis: (A) Immunostaining is moderate in the small (s) and intense in pyriform cells (p), a weak staining is detected in the ooplasm (o). (B) Intense ERα immunostaining in granules on the pyriform cells (p) and cortex (c).

Therefore, we studied the previtellogenic follicles obtained in October that showed a weak mark in the cytoplasm of the granulosa and ooplasm with a strong localization in some granules close to the cortex and some pyriform cells granules (Figure 3A). In addition, the ERβ was strong in the oocyte cortex, erythrocytes, and moderate in granulosa and theca (Figure 3D). For PR, the stain was intense in the pyriform cells and moderate in the oocyte cortex and some small cells (Figure 3G). With the beginning of vitellogenesis, the ERα staining was strong in the erythrocytes located in the blood vessels and weak in the oocyte cortex (Figure 3B). Conversely, ERβ was moderate in theca, granulosa cells, and oocyte’s cortex (Figure 3E). The PR mark was strong during pyriform cells regression, and moderate in the oocyte cortex and cytoplasm of small cells (Figure 3H).

Figure 3. Immunolocalization of ERα (A, B, C), ERβ (D, E, F) and PR (G, H, I) in the ovarian follicles. Previtellogenesis: (A) Intense ERα immunostaining in granules on the pyriform cells (p) and cortex (c). (D) Strong ERβ immunostaining in cortex (c), erythrocytes located in the blood vessels (v) and moderate in granulosa (p, s) and theca (t). (G) Strong immunolabelling of PR in pyriform cells (p), moderate in oocyte cortex (c) and small cells (s). Vitellogenesis: (B) ERα intense immunostaining in erythrocytes located in the blood vessels (v) and weak in cortex (c). (E) ERβ was moderate in theca (t), granulosa cells (s, p) and oocyte cortex (c). (H) Strong PR immunolabelling in regressed pyriform cells (p) and moderate in the oocyte cortex (c) and small cells (s). Preovulatory: (C) Intense ERα immunoreactivity in the cortex (c) and theca (t). (F) No evident ERβ expression. (I) Moderate PR in the yolk (y). (J, K, L) omission controls.

In the preovulatory stage, there is an evident reduction in the immunostaining of the three receptors. The ERα was intense in the cortex and some regions of the theca (Figure 3C). There was no ERβ expression in this stage (Figure 3F). Finally, the presence of PR was moderate in yolk (Figure 3I).

Discussion

Sex steroid receptors are involved in multiple events in the reproductive biology of female vertebrates. They have different patterns of expression depending on the tissue and the reproductive status of the organism (Hammes and Levin, Reference Hammes and Levin2007; Jacobsen and Horwitz, Reference Jacobsen and Horwitz2012; Baker, Reference Baker2019; Fuentes and Silveyra, Reference Fuentes and Silveyra2019). The interaction of these hormones with their receptors is necessary to activate the expression of one or multiple genes that act on the phenotype (Cox, Reference Cox2020). As they have cytoplasmic and nuclear localization in the cells of many tissues (Dressing et al., Reference Dressing, Goldberg, Charles, Schwertfeger and Lange2011), they show high-functional plasticity due to their large number of combinations in modulators, levels of activity, and responses (Jacobsen and Horwitz, Reference Jacobsen and Horwitz2012). Numerous studies have reported their participation in the reproductive biology of female vertebrates (Hamilton et al., Reference Hamilton, Hewitt, Arao, Korach, Forrest and Tsai2017; Baker, Reference Baker2019; Fuentes and Silveyra, Reference Fuentes and Silveyra2019). However, there is limited information concerning their presence in lizards. In this study, we observed immunolocalization of ERα, Erβ, and PR in previtellogenic, vitellogenic, and preovulatory follicles of S. torquatus.

The ovarian follicles showed changes in their morphology as reported for other lizards (Uribe et al., Reference Uribe, Omana, Quintero and Guillette1995; Raucci and di Fiore, Reference Raucci and di Fiore2010; Motta et al., Reference Motta, Tammaro, Di Lorenzo, Panzuto, Verderame, Migliaccio and Simoniello2020). The more evident changes were the modification of the granulosa layer and the deposit of yolk inside the oocyte. The expression of steroid receptors changed according to the stage of follicular development and the cellular types registered. E2 concentrations were significantly higher in the preovulatory stage. In addition, P4 showed similar levels among the different phases of follicular development. The same behaviour is reported for Sceloporus virgatus (Weiss et al., Reference Weiss, Jennings and Moore2002), Phrynosoma cornutum (Wack et al., Reference Wack, Fox, Hellgren and Lovern2008), Chameleo calyptratus (Kummrow et al., Reference Kummrow, Smith, Crawshaw and Mastromonaco2010), Salvator merianae (Zena et al., Reference Zena, Dillon, Hunt, Navas, Bícego and Buck2019) and Mabuya sp. (Barbosa-Moyano et al., Reference Barbosa-Moyano, Rodríguez-Chaparro, Santos and Ramírez-Pinilla2020). ER, and PR presence was not related to the sex steroid concentrations, as reported in the lizard U. acanthinura (Hammouche et al., Reference Hammouche, Gernigon and Exbrayat2017).

We found that the ER labelling was mainly expressed in the granulosa layer and oocyte cortex. In the previtellogenic phase, ERα in the small cells could be associated with the proliferative activity to differentiate in a polymorphic granulosa (Kocanova et al., Reference Kocanova, Mazaheri, Caze-Subra and Bystricky2010; Hamilton et al., Reference Hamilton, Hewitt, Arao, Korach, Forrest and Tsai2017). Also, its presence in the cytoplasm of the pyriform cells may indicate their role in the estrogen stimulation synthesis and secretion in previtellogenic follicles (Hammouche et al., Reference Hammouche, Gernigon and Exbrayat2017; Fuentes and Silveyra, Reference Fuentes and Silveyra2019), inducing aromatase activity at the beginning of the reproductive season (Tripathy and Rai, Reference Tripathy and Rai2017). This steroidogenic capacity has been observed in the granulosa cells of previtellogenic and early vitellogenic follicles in S. torquatus (unpublished data).

For ERβ, its hepatic expression remained constant in the reproductive cycle of the lizard P. sicula (Verderame and Limatola, Reference Verderame and Limatola2010) and changed in the ovaries of H. flaviviridis (Tripathy and Rai, Reference Tripathy and Rai2017). Both receptors’ presence may be involved with the regression of pyriform cells in S. torquatus during vitellogenesis, as we observed an increased expression in atretic follicles.

Although previous studies have reported the presence of sex steroid receptors in diverse tissues on lizards (Young et al., Reference Young, Lopreato, Horan and Crews1994; Beck and Wade, Reference Beck and Wade2009; Verderame and Limatola, Reference Verderame and Limatola2010; Hammouche et al., Reference Hammouche, Remana and Exbrayat2012; Inamdar et al., Reference Inamdar, Khodnapur, Nindi, Dasari and Seshagiri2015; Yatsu et al., Reference Yatsu, Katsu, Kohno, Mizutani, Ogino, Ohta, Myburgh, van Wyk, Guillette, Miyagawa and Iguchi2016), there is limited information available on this group. as their expression was mainly cytosolic, further studies must consider signalling via non-genomic mechanisms. ER presence in the cytosol is associated with an unbound state that allows the translocation between the nucleus and cytoplasm (Kocanova et al., Reference Kocanova, Mazaheri, Caze-Subra and Bystricky2010).

The expression of PR in S. torquatus showed a different spatial-temporal pattern expression. However, identifying the behaviour of each isoform could shed light on its role in the phases of follicular development. In reptiles, both isoforms have been reported and are associated with the stage of the reproductive cycle (Custodia-Lora and Callard, Reference Custodia-Lora and Callard2002; Biazik et al., Reference Biazik, Parker, Murphy and Thompson2012; Hammouche et al., Reference Hammouche, Remana and Exbrayat2012; Motta et al., Reference Motta, Tammaro, Di Lorenzo, Panzuto, Verderame, Migliaccio and Simoniello2020). During follicular development, PR could stimulate the proliferation in the germinal beds, recruitment, and transition to the primary oocyte (Custodia-Lora and Callard, Reference Custodia-Lora and Callard2002; Jones, Reference Jones, Norris and Lopez2011; Holding et al., Reference Holding, Frazier, Dorr, Pollock, Muelleman, Branske, Henningsen, Eikenaar, Escallón, Montgomery, Moore and Taylor2014; Ramírez-Pinilla et al., Reference Ramírez-Pinilla, de Pérez, Alvarado-Ramírez, Rheubert, Siegel and Trauth2015; Duarte-Méndez et al., Reference Duarte-Méndez, Quintero-Silva and Ramírez-Pinilla2018; Motta et al., Reference Motta, Tammaro, Di Lorenzo, Panzuto, Verderame, Migliaccio and Simoniello2020). As the follicular development progressed, we observed an increase in immunolabelling intensity. A similar effect was observed in Uromastix acanthinura, where E2 positively influenced PRA and PRB (Shao et al., Reference Shao, Markström, Friberg, Johansson and Billig2003; Hammouche et al., Reference Hammouche, Remana and Exbrayat2012). In the turtle Chrysems picta, PRA expression inhibits VTG synthesis (Duggan and Callard, Reference Duggan and Callard2003) whereas PRB promotes it (Giannoukos and Callard, Reference Giannoukos and Callard1995). In the preovulatory stage, PR presence can regulate ovulation and caspase 3 activity (Shao et al., Reference Shao, Markström, Friberg, Johansson and Billig2003; Hammouche et al., Reference Hammouche, Gernigon and Exbrayat2017). Therefore, the differences in the events may be mediated by each receptor in the different species.

Our observations indicate that granulosa cells can have different roles in oocyte maturation, depending on the sensitivity to diverse intraovarian factors. The immunolabelling of the sex steroid receptors studied in the granulosa cells and, particularly the pyriform cells, have similar behaviour as other receptors influencing gonadotropin secretion of bradykinin, c-kit, and stem cell factor (Singh et al., Reference Singh, Krishna, Sridaran and Tsutsui2008; Raucci and di Fiore, Reference Raucci and di Fiore2011). The above confirms their support role in transferring diverse components to the oocyte after their breakdown (Neaves, Reference Neaves1971; Taddei, Reference Taddei1972; Filosa and Taddei, Reference Filosa and Taddei1976; Andreuccetti, Reference Andreuccetti1992; Motta et al., Reference Motta, Scanderbeg and Filosa1995; Aldokhi et al., Reference Aldokhi, Alwasel and Harrath2019).

In conclusion, our study revealed that the immunolabelling of ER and PR was more pronounced in the pyriform cells and oocyte cortex. These observations suggest that in females of S. torquatus, steroid receptors participate in the regulation of follicular development as in other vertebrates. Those receptors have a spatio-temporal expression depending on the stage of the reproductive cycle.

Acknowledgements

This work was supported by the Consejo Nacional de Ciencia y Tecnología (CONACYT) through a scholarship awarded to N. B. Cruz-Cano (CVU 621940). We are grateful to the Sierra de Guadalupe State Park staff, Juan Gerardo Valverde (regional delegate of Tultitlán) and Mario Alberto Serrano Pérez (Protection and Surveillance Assistant). Also, we thank the field assistance of Romeo Eduardo Loya-Zurita, Yabín Josué Castro-Camacho, Alejandro Cadena Velázquez, Julián Torres Gloria and Rodrigo Dávila-Govantes. For the manuscript review to David Osvaldo Cruz-Cano. We express our gratitude to the anonymous reviewers.

Financial support

This work was supported by the CONACYT through a scholarship awarded to N. B. Cruz-Cano (CVU 621940).

Competing interest

None

Ethical standard

The UNAM Bioethical Committee approved all experimental procedures.

References

Adams, S. M., Biazik, J., Stewart, R. L., Murphy, C. R. and Thompson, M. B. (2007). Fundamentals of viviparity: Comparison of seasonal changes in the uterine epithelium of oviparous and viviparous Lerista bougainvillii (Squamata: Scincidae). Journal of Morphology, 268(7), 624–635. doi: 10.1002/jmor.10522 CrossRefGoogle ScholarPubMed
Al-Amri, I. S., Mahmoud, I. Y., Waring, C. P., Alkindi, A. Y., Khan, T. and Bakheit, C. (2012). Seasonal changes in plasma steroid levels in relation to ovarian steroidogenic ultrastructural features and progesterone receptors in the house gecko, Hemidactylus flaviviridis, in Oman. General and Comparative Endocrinology, 177(1), 46–54. doi: 10.1016/j.ygcen.2012.02.006 CrossRefGoogle ScholarPubMed
Aldokhi, O. A., Alwasel, S. and Harrath, A. H. (2019). Ultrastructural and histochemical study of previtellogenic oogenesis in the desert lizard Scincus mitranus (Squamata, Sauropsida). Journal of Morphology, 280(3), 381–394. doi: 10.1002/jmor.20950 CrossRefGoogle ScholarPubMed
Amey, A. P. and Whittier, J. M. (2000). Seasonal patterns of plasma steroid hormones in males and females of the bearded dragon lizard, Pogona barbata . General and Comparative Endocrinology, 117(3), 335–342. doi: 10.1006/gcen.2000.7426 CrossRefGoogle ScholarPubMed
Andreuccetti, P. (1992). An ultrastructural study of differentiation of pyriform cells and their contribution to oocyte growth in representative Squamata. Journal of Morphology, 212(1), 1–11. doi: 10.1002/jmor.1052120102 CrossRefGoogle ScholarPubMed
Baker, M. E. (2019). Steroid receptors and vertebrate evolution. Molecular and Cellular Endocrinology, 496, 110526. doi: 10.1016/j.mce.2019.110526 CrossRefGoogle ScholarPubMed
Barbosa-Moyano, H., Rodríguez-Chaparro, S., Santos, R. and Ramírez-Pinilla, M. (2020). Plasma estradiol and progesterone concentrations during the female reproductive cycle in a highly placentotrophic viviparous lizard, Mabuya sp. General and Comparative Endocrinology, 295, 113530. doi: 10.1016/j.ygcen.2020.113530 CrossRefGoogle Scholar
Beck, L. A. and Wade, J. (2009). Sexually dimorphic estrogen receptor α mRNA expression in the preoptic area and ventromedial hypothalamus of green anole lizards. Hormones and Behavior, 55(3), 398–403. doi: 10.1016/j.yhbeh.2009.01.003 CrossRefGoogle ScholarPubMed
Biazik, J. M., Parker, S. L., Murphy, C. R. and Thompson, M. B. (2012). Uterine epithelial morphology and progesterone receptors in a mifepristone-treated viviparous lizard Pseudemoia entrecasteauxii (Squamata: Scincidae) during gestation. Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution, 318(2), 148–158. doi: 10.1002/jez.b.22003 CrossRefGoogle Scholar
Blackburn, D. G. (2018). Reproduction in reptiles. In Skinner, M. K. (ed.) Encyclopedia of Reproduction 2nd edition vol. 6, pp. 573–578. Elsevier. doi: 10.1016/b978-0-12-809633-8.20651-1 CrossRefGoogle Scholar
Bonnet, X., Naulleau, G., Bradshaw, D. and Shine, R. (2001). Changes in plasma progesterone in relation to vitellogenesis and gestation in the viviparous snake Vipera aspis . General and Comparative Endocrinology, 121(1), 84–94. doi: 10.1006/gcen.2000.7574 CrossRefGoogle ScholarPubMed
Contrò, V., Basile, J. R. and Proia, P. (2015). Sex steroid hormone receptors, their ligands, and nuclear and non-nuclear pathways. AIMS Molecular Science, 2(3), 294–310. doi: 10.3934/molsci.2015.3.294 CrossRefGoogle Scholar
Cox, R. M. (2020). Sex steroids as mediators of phenotypic integration, genetic correlations, and evolutionary transitions. Molecular and Cellular Endocrinology, 502, 110668. doi: 10.1016/j.mce.2019.110668 CrossRefGoogle ScholarPubMed
Crews, D. (2005). Evolution of neuroendocrine mechanisms that regulate sexual behavior. Trends in Endocrinology and Metabolism, 16(8), 354–361. doi: 10.1016/j.tem.2005.08.007 CrossRefGoogle ScholarPubMed
Cruz-Cano, N. B., Sánchez-Rivera, U. Á., Álvarez-Rodríguez, C., Dávila-Govantes, R., Cárdenas-León, M. and Martínez-Torres, M. (2021). Sex steroids are correlated with environmental factors and body condition during the reproductive cycle in females of the lizard Sceloporus torquatus . General and Comparative Endocrinology, 314, 113921. doi: 10.1016/j.ygcen.2021.113921 CrossRefGoogle ScholarPubMed
Custodia-Lora, N. and Callard, I. P. (2002). Progesterone and progesterone receptors in reptiles. General and Comparative Endocrinology, 127(1), 1–7. doi: 10.1016/S0016-6480(02)00030-8 CrossRefGoogle ScholarPubMed
Custodia-Lora, N., Novillo, A. and Callard, I. P. (2004). Regulation of hepatic progesterone and estrogen receptors in the female turtle, Chrysemys picta: Relationship to vitellogenesis. General and Comparative Endocrinology, 136(2), 232–240. doi: 10.1016/j.ygcen.2003.12.016 CrossRefGoogle ScholarPubMed
Dressing, G. E., Goldberg, J. E., Charles, N. J., Schwertfeger, K. L. and Lange, C. A. (2011). Membrane progesterone receptor expression in mammalian tissues: A review of regulation and physiological implications. Steroids, 76(1–2), 11–17. doi: 10.1016/j.steroids.2010.09.006 CrossRefGoogle ScholarPubMed
Duarte-Méndez, M., Quintero-Silva, J. and Ramírez-Pinilla, M. P. (2018). Immunohistochemical localization of 3β-hydroxysteroid dehydrogenase and progesterone receptors in the ovary and placenta during gestation of the placentotrophic lizard Mabuya sp. (Squamata: Scincidae). General and Comparative Endocrinology, 261, 136–147. doi: 10.1016/j.ygcen.2018.02.017 CrossRefGoogle ScholarPubMed
Duggan, A. E. and Callard, I. P. (2003). Lipids and lipid-transporting proteins in Chrysemys picta: Role of gonadal steroids and growth hormone in intact and hypophysectomized turtles. General and Comparative Endocrinology, 131(2), 176–184. doi: 10.1016/s0016-6480(03)00009-1 CrossRefGoogle ScholarPubMed
Edwards, D. P. (2005). Regulation of signal transduction pathways by estrogen and progesterone. Annual Review of Physiology, 67, 335–376. doi: 10.1146/annurev.physiol.67.040403.120151 CrossRefGoogle ScholarPubMed
Edwards, A. and Jones, S. M. (2001). Changes in plasma progesterone, estrogen, and testosterone concentrations throughout the reproductive cycle in female viviparous blue-tongued skinks, Tiliqua nigrolutea (Scincidae), in Tasmania. General and Comparative Endocrinology, 122(3), 260–269. doi: 10.1006/gcen.2001.7634 CrossRefGoogle ScholarPubMed
Filosa, S. and Taddei, C. (1976). Intercellular bridges in lizard oogenesis. Cell Differentiation, 5(3), 199–206. doi: 10.1016/0045-6039(76)90021-x CrossRefGoogle ScholarPubMed
Fuentes, N. and Silveyra, P. (2019). Estrogen receptor signaling mechanisms. Advances in Protein Chemistry and Structural Biology, 116, 135–170. doi: 10.1016/bs.apcsb.2019.01.001 CrossRefGoogle ScholarPubMed
Garg, D., Ng, S. S. M., Baig, K. M., Driggers, P. and Segars, J. (2017). Progesterone-mediated non-classical signaling. Trends in Endocrinology and Metabolism, 28(9), 656–668. doi: 10.1016/j.tem.2017.05.006 CrossRefGoogle ScholarPubMed
Giannoukos, G. and Callard, I. P. (1995). Reptilian (Chrysemys picta) hepatic progesterone receptors: Relationship to plasma steroids and the vitellogenic cycle. Journal of Steroid Biochemistry and Molecular Biology, 55(1), 93–106. doi: 10.1016/0960-0760(95)00149-t CrossRefGoogle ScholarPubMed
Girling, J. E., Jones, S. M. and Swain, R. (2002). Delayed ovulation and parturition in a viviparous alpine lizard (Niveoscincus microlepidotus): Morphological data and plasma steroid concentrations. Reproduction, Fertility, and Development, 14(1–2), 43–53. doi: 10.1071/RD01091 CrossRefGoogle Scholar
Guillette, L. J. and la Cruz, F. R. M. (1993). The reproductive cycle of the viviparous Mexican lizard Sceloporus torquatus . Journal of Herpetology, 27(2), 168–174. doi: 10.2307/1564933 CrossRefGoogle Scholar
Hamilton, K. J., Hewitt, S. C., Arao, Y. and Korach, K. S. (2017). Estrogen hormone biology. In: Forrest, D. and Tsai, S. (eds). Current Topics in Developmental Biology 125, 109–146. Academic Press, Inc. doi: 10.1016/bs.ctdb.2016.12.005 Google Scholar
Hammes, S. R. and Levin, E. R. (2007). Extranuclear steroid receptors: Nature and actions. Endocrine Reviews, 28(7), 726–741. doi: 10.1210/er.2007-0022 CrossRefGoogle ScholarPubMed
Hammouche, S. B., Remana, S. and Exbrayat, J. M. (2012). Immunolocalization of hepatic estrogen and progesterone receptors in the female lizard Uromastyx acanthinura . Comptes Rendus Biologies, 335(7), 445–453. doi: 10.1016/j.crvi.2012.06.002 CrossRefGoogle ScholarPubMed
Hammouche, S., Gernigon, T. and Exbrayat, J. (2017). Immunolocalization of estrogens and progesterone receptors within the ovary of the lizard Uromastyx acanthinura from vitellogenesis to rest season. Folia Histochemica et Cytobiologica, 45, 23–27.Google Scholar
Holding, M. L., Frazier, J. A., Dorr, S. W., Pollock, N. B., Muelleman, P. J., Branske, A., Henningsen, S. N., Eikenaar, C., Escallón, C., Montgomery, C. E., Moore, I. T. and Taylor, E. N. (2014). Wet- and dry-season steroid hormone profiles and stress reactivity of an insular dwarf snake, the hog island boa (Boa constrictor imperator). Physiological and Biochemical Zoology, 87(3), 363–373. doi: 10.1086/675938 CrossRefGoogle ScholarPubMed
Holmes, K. M. and Cree, A. (2006). Annual reproduction in females of a viviparous skink (Oligosoma maccanni) in a subalpine environment. Journal of Herpetology, 40(2), 141–151. doi: 10.1670/235-04.1 CrossRefGoogle Scholar
Hoss, S. K., Garcia, M. J., Earley, R. L. and Clark, R. W. (2014). Fine-scale hormonal patterns associated with birth and maternal care in the cottonmouth (Agkistrodon piscivorus), a North American pitviper snake. General and Comparative Endocrinology, 208, 85–93. doi: 10.1016/j.ygcen.2014.08.011 CrossRefGoogle ScholarPubMed
Inamdar, L. S., Khodnapur, B. S., Nindi, R. S., Dasari, S. and Seshagiri, P. B. (2015). Differential expression of estrogen receptor alpha in the embryonic adrenal-kidney-gonadal complex of the oviparous lizard, Calotes versicolor (Daud.). General and Comparative Endocrinology, 220, 55–60. doi: 10.1016/j.ygcen.2014.08.003 CrossRefGoogle ScholarPubMed
Jacobsen, B. M. and Horwitz, K. B. (2012). Progesterone receptors, their isoforms and progesterone regulated transcription. Molecular and Cellular Endocrinology, 357(1–2), 18–29. doi: 10.1016/j.mce.2011.09.016 CrossRefGoogle ScholarPubMed
Jones, S. M. (2011). Hormonal regulation of ovarian function in reptiles. In: Norris, D. O. and Lopez, K. H. (eds). Hormones and Reproduction of Vertebrates, vol. 3, pp. 89–115. Elsevier, Inc. doi: 10.1016/B978-0-12-374930-7.10004-4 Google Scholar
Kabelik, D., Weiss, S. L. and Moore, M. C. (2008). Steroid hormones alter neuroanatomy and aggression independently in the tree lizard. Physiology and Behavior, 93(3), 492–501. doi: 10.1016/j.physbeh.2007.10.008 CrossRefGoogle ScholarPubMed
Kocanova, S., Mazaheri, M., Caze-Subra, S. and Bystricky, K. (2010). Ligands specify estrogen receptor alpha nuclear localization and degradation. BMC Cell Biology, 11, 98. doi: 10.1186/1471-2121-11-98 CrossRefGoogle ScholarPubMed
Kummrow, M. S., Smith, D. A., Crawshaw, G. and Mastromonaco, G. F. (2010). Characterization of fecal hormone patterns associated with the reproductive cycle in female veiled chameleons (Chamaeleo calyptratus). General and Comparative Endocrinology, 168(3), 340–348. doi: 10.1016/j.ygcen.2010.04.022 CrossRefGoogle ScholarPubMed
Lovern, M. B. (2011). Hormones and reproductive cycles in lizards. In: Norris, D. O. and Lopez, K. H. (eds). Hormones and Reproduction of Vertebrates, vol. 3, pp. 321–353. Elsevier, Inc. doi: 10.1016/B978–0-12–374930–7.10012–3 Google Scholar
Martínez-Torres, M., Elena Hernández-Caballero, M., Alvarez-Rodriguez, C., Alba Luis-Díaz, J. and Ortíz-López, G. (2003). Luteal development and progesterone levels during pregnancy of the viviparous temperate lizard Barisia imbricata imbricata (Reptilia: Anguidae). General and Comparative Endocrinology, 132(1), 55–65. doi: 10.1016/s0016-6480(02)00607-x CrossRefGoogle ScholarPubMed
Medina-Laver, Y., Rodríguez-Varela, C., Salsano, S., Labarta, E. and Domínguez, F. (2021). What do we know about classical and non-classical progesterone receptors in the human female reproductive tract? A review. International Journal of Molecular Sciences, 22(20). doi: 10.3390/ijms222011278 CrossRefGoogle ScholarPubMed
Motta, C. M., Scanderbeg, M. C. and Filosa, S. (1995). Role of pyriform cells during the growth of oocytes in the lizard Podarcis sicula . Journal of Experimental Zoology, 256, 247–256.CrossRefGoogle Scholar
Motta, C. M., Tammaro, S., de Stasio, R., Borrelli, L. and Filosa, S. (2004). How follicle number is regulated in the ovary of the lizard Podarcis sicula? Italian Journal of Zoology, 71 sup2, 109–111. doi: 10.1080/11250000409356618 CrossRefGoogle Scholar
Motta, C. M., Tammaro, S., Di Lorenzo, M., Panzuto, R., Verderame, M., Migliaccio, V. and Simoniello, P. (2020). Spring and fall recrudescence in Podarcis siculus ovaries: A role for progesterone. General and Comparative Endocrinology, 290, 113393. doi: 10.1016/j.ygcen.2020.113393 CrossRefGoogle ScholarPubMed
Neaves, W. B. (1971). Intercellular bridges between follicle cells and oocyte in the lizard, Anolis carolinensis . Anatomical Record, 170(3), 285–301. doi: 10.1002/ar.1091700305 CrossRefGoogle ScholarPubMed
Norris, D. O. (2018). Comparative endocrinology: Past, present, and future. Integrative and Comparative Biology, 58(6), 1033–1042. doi: 10.1093/icb/icy107 Google ScholarPubMed
Ortiz, M. F., de Oca, A. N., Ugarte, I. H. S., de Oca, A. N. and Ugarte, I. H. S. (2001). Diet and reproductive biology of the viviparous lizard Sceloporus torquatus torquatus (Squamata: Phrynosomatidae). Journal of Herpetology, 35(1), 104–112. doi: 10.2307/1566029 CrossRefGoogle Scholar
Ramírez-Pinilla, M. P., de Pérez, G. R. and Alvarado-Ramírez, C. (2015). Oogenesis and the ovarian cycle. In: Rheubert, J. L., Siegel, D. S. & Trauth, S. E. (eds). Reproductive Biology and Phylogeny of Lizards and Tuatara. Chapter 8. CRC Press.Google Scholar
Raucci, F. and di Fiore, M. M. (2010). The maturation of oocyte follicular epithelium of Podarcis s. sicula is promoted by D-aspartic acid. Journal of Histochemistry and Cytochemistry, 58(2), 157–171. doi: 10.1369/jhc.2009.954636 CrossRefGoogle ScholarPubMed
Raucci, F. and di Fiore, M. M. (2011). Localization of c-kit and stem cell factor (SCF) in ovarian follicular epithelium of a lizard, Podarcis s. sicula . Acta Histochemica, 113(6), 647–655. doi: 10.1016/j.acthis.2010.08.004 CrossRefGoogle ScholarPubMed
Refsnider, J. M., Clifton, I. T. and Vazquez, T. K. (2019). Developmental plasticity of thermal ecology traits in reptiles: Trends, potential benefits, and research needs. Journal of Thermal Biology, 84, 74–82. doi: 10.1016/j.jtherbio.2019.06.005 CrossRefGoogle ScholarPubMed
Rhen, T., Sakata, J. T., Zeller, M. and Crews, D. (2000). Sex steroid levels across the reproductive cycle of female leopard geckos, Eublepharis macularius, from different incubation temperatures. General and Comparative Endocrinology, 118(2), 322–331. doi: 10.1006/gcen.2000.7477 CrossRefGoogle ScholarPubMed
Sánchez, F. and Smitz, J. (2012). Molecular control of oogenesis. Biochimica et Biophysica Acta, 1822(12), 1896–1912. doi: 10.1016/j.bbadis.2012.05.013 CrossRefGoogle ScholarPubMed
Sato, T., Miyagawa, S. and Iguchi, T. (2016). Progesterone. In: Takei, Y., Ando, H. & Tsutsui, K. (eds) Handbook of Hormones pp. 507–e94A-3. Elsevier. doi: 10.1016/B978-0-12-801028-0.00220-8 CrossRefGoogle Scholar
Shao, R., Markström, E., Friberg, P. A., Johansson, M. and Billig, H. (2003). Expression of progesterone receptor (PR) A and B isoforms in mouse granulosa cells: Stage-dependent PR-mediated regulation of apoptosis and cell proliferation. Biology of Reproduction, 68(3), 914–921. doi: 10.1095/biolreprod.102.009035 CrossRefGoogle Scholar
Singh, P., Krishna, A., Sridaran, R. and Tsutsui, K. (2008). Changes in GnRH I, bradykinin and their receptors and GnIH in the ovary of Calotes versicolor during reproductive cycle. General and Comparative Endocrinology, 159(2–3), 158–169. doi: 10.1016/j.ygcen.2008.08.016 CrossRefGoogle ScholarPubMed
Taddei, C. (1972). Significance of pyriform cells in ovarian follicle of Lacerta sicula . Experimental Cell Research, 72(2), 562–566. doi: 10.1016/0014-4827(72)90031-6 CrossRefGoogle ScholarPubMed
Tammaro, S., Simoniello, P., Filosa, S. and Motta, C. M. (2008). cGnRH II involvement in pyriform cell apoptosis. Cell and Tissue Research, 332(2), 337–347. doi: 10.1007/s00441-008-0584-x CrossRefGoogle ScholarPubMed
Tripathy, M. and Rai, U. (2017). Temporal expression and gonadotropic regulation of aromatase and estrogen receptors in the ovary of wall lizard, Hemidactylus flaviviridis: Correlation with plasma estradiol and ovarian follicular development. Steroids, 128, 23–31. doi: 10.1016/j.steroids.2017.10.005 CrossRefGoogle ScholarPubMed
Uribe, M. D. C. A., Omana, M. E. M., Quintero, J. G. and Guillette, L. J. Jr (1995). Seasonal variation in ovarian histology of the viviparous lizard Sceloporus torquatus torquatus . Journal of Morphology, 226(1), 103–119. doi: 10.1002/jmor.1052260107 CrossRefGoogle ScholarPubMed
Verderame, M. and Limatola, E. (2010). Molecular identification of estrogen receptors (ERα and ERβ) and their differential expression during VTG synthesis in the liver of lizard Podarcis sicula . General and Comparative Endocrinology, 168(2), 231–238. doi: 10.1016/j.ygcen.2010.04.014 CrossRefGoogle ScholarPubMed
Wack, C. L., Fox, S. F., Hellgren, E. C. and Lovern, M. B. (2008). Effects of sex, age, and season on plasma steroids in free-ranging Texas horned lizards (Phrynosoma cornutum). General and Comparative Endocrinology, 155(3), 589–596. doi: 10.1016/j.ygcen.2007.10.005 CrossRefGoogle ScholarPubMed
Weiss, S. L., Jennings, D. H. and Moore, M. C. (2002). Effect of captivity in semi-natural enclosures on the reproductive endocrinology of female lizards. General and Comparative Endocrinology, 128(3), 238–246. doi: 10.1016/s0016-6480(02)00506-3 CrossRefGoogle ScholarPubMed
Yaşar, P., Ayaz, G., User, S. D., Güpür, G. and Muyan, M. (2017). Molecular mechanism of estrogen–estrogen receptor signaling. Reproductive Medicine and Biology, 16(1), 4–20. doi: 10.1002/rmb2.12006 CrossRefGoogle ScholarPubMed
Yatsu, R., Katsu, Y., Kohno, S., Mizutani, T., Ogino, Y., Ohta, Y., Myburgh, J., van Wyk, J. H., Guillette, L. J., Miyagawa, S. and Iguchi, T. (2016). Characterization of evolutionary trend in squamate estrogen receptor sensitivity. General and Comparative Endocrinology, 238, 88–95. doi: 10.1016/j.ygcen.2016.04.005 CrossRefGoogle ScholarPubMed
Young, L. J., Lopreato, G. F., Horan, K. and Crews, D. (1994). Cloning and in situ hybridization analysis of estrogen receptor, progesterone receptor, and androgen receptor expression in the brain of whiptail lizards (Cnemidophorus uniparens and C. inornatus). Journal of Comparative Neurology, 347(2), 288–300. doi: 10.1002/cne.903470210 CrossRefGoogle ScholarPubMed
Zena, L. A., Dillon, D., Hunt, K. E., Navas, C. A., Bícego, K. C. and Buck, C. L. (2019). Seasonal changes in plasma concentrations of the thyroid, glucocorticoid and reproductive hormones in the tegu lizard Salvator merianae . General and Comparative Endocrinology, 273, 134–143. doi: 10.1016/j.ygcen.2018.06.006 CrossRefGoogle ScholarPubMed
Figure 0

Figure 1. Morphology of ovarian follicles in the lizard S. torquatus. Previtellogenesis: (A) The follicles have a homogeneous ooplasm (o) with a cortex in the oocyte periphery (c); the granulosa is a multilayered structure with polymorphic cells small (s), intermediate (i) and pyriform (p). The zona pellucida (z) and a fibrous theca (t) with blood vessels (v) are clearly distinguishable. Vitellogenesis: (B) The oocyte contains yolk granules (y), and the granulosa is comprised by a monolayer of small cells (s), the zona pellucida (z) is thicker, the theca (t) has blood vessels (v) and mast cells (mc), and the small cells (s) begin to flatten. Preovulatory: (C) The zona pellucida (z) decreases in thickness and the small cells (s) are flattened in a single layer, also there is a decrease in the blood vessels (v). (A–C Masson’s Trichrome stained.)

Figure 1

Table 1. Plasma sex steroid concentrations in different phases of the follicular development in the lizard S. torquatus

Figure 2

Figure 2. Immunolocalization of ERα in atretic previtellogenic ovarian follicles. Previtellogenesis: (A) Immunostaining is moderate in the small (s) and intense in pyriform cells (p), a weak staining is detected in the ooplasm (o). (B) Intense ERα immunostaining in granules on the pyriform cells (p) and cortex (c).

Figure 3

Figure 3. Immunolocalization of ERα (A, B, C), ERβ (D, E, F) and PR (G, H, I) in the ovarian follicles. Previtellogenesis: (A) Intense ERα immunostaining in granules on the pyriform cells (p) and cortex (c). (D) Strong ERβ immunostaining in cortex (c), erythrocytes located in the blood vessels (v) and moderate in granulosa (p, s) and theca (t). (G) Strong immunolabelling of PR in pyriform cells (p), moderate in oocyte cortex (c) and small cells (s). Vitellogenesis: (B) ERα intense immunostaining in erythrocytes located in the blood vessels (v) and weak in cortex (c). (E) ERβ was moderate in theca (t), granulosa cells (s, p) and oocyte cortex (c). (H) Strong PR immunolabelling in regressed pyriform cells (p) and moderate in the oocyte cortex (c) and small cells (s). Preovulatory: (C) Intense ERα immunoreactivity in the cortex (c) and theca (t). (F) No evident ERβ expression. (I) Moderate PR in the yolk (y). (J, K, L) omission controls.