The Involvement of the Androgen Receptor in the Secretion of the Epididymalcorpusin the LizardPodarcis sicula

A crucial role in the maintenance of male reproductive functions is carried out by the androgen through its receptor in balance with the estrogen receptors (ERs). The distribution of the androgen receptor (AR) is well documented in the testis and in the reproductive tissues of mammals but the findings about the AR in nonmammalian vertebrates and in particular in reptiles are very scarce. Here by means ofin situhybridization (ISH) we investigated the AR expression along the epididymal channel (efferent ductules,corpus, andcauda) ofPodarcis siculaduring the mating and nonmating period. The results show that in this seasonal breeding species the AR expression pattern is always constant throughout the epididymis. The administration of estradiol-17βin the mating period does not affect the AR expression but inhibits the secretory activity of the epididymalcorpus. To verify the expression pattern of ERs, we also conducted ISH investigations on adjacent sections with ERs probes. The findings suggest that AR induces the secretory activity in the epithelial cells of the epididymalcorpusand confirm our previous results that showed the role of ERalpha (ERα) as switch off for the secretion of this compartment

Molecular and Histological Effects of Glyphosate on Testicular Tissue of the Lizard Podarcis siculus

The expansion of agriculture produces a steady increase in habitat fragmentation and deg‐ radation due to the increased use of pesticides and herbicides. Habitat loss and alteration associated with crop production play an important role in reptile decline, among which lizards are particularly endangered. In this study, we evaluated testicular structure, steroidogenesis, and estrogen receptor expression/localization after three weeks of oral exposure to glyphosate at 0.05 and 0.5 μg/kg body weight every other day in the field lizard Podarcis siculus. Our results show that glyphosate affected testicular morphology, reduced spermatogenesis, altered gap junctions and changed the localiza‐ tion of estrogen receptors in germ cells, increasing their expression; the effects were mostly dose‐ dependent. The result also demonstrates that glyphosate, at least at these concentrations, did not influence steroidogenesis. Overall, the data indicate that this herbicide can disturb the morphophys‐ iology of the male lizard’s reproductive system, with obviously detrimental effects on their repro‐ ductive fitness. The effects of glyphosate must be considered biologically relevant and could endan‐ ger the reproductive capacity not only of lizards but also of other vertebrates, including humans; a more controlled and less intensive use of glyphosate in areas devoted to crop production would therefore be advisable

Exploring the Role of Estrogens in Lizard Spermatogenesis through the Study of Clomiphene and FSH Effects

Spermatogenesis is a fascinating biological process aiming to generate haploid spermatozoa from diploid spermatogonia through a specific hormonal network between gonadotropins and steroids. Increasing evidence suggests that the primary female sex hormone estrogen plays an active role in this process. This research points out on the role of estrogen during lizard spermatogenesis by using three experimental approaches: (1) exposure to an analogue of nonsteroidal estrogen as Clomiphene citrate that acts both as estrogen agonist and antagonist; (2) exposure to the gonadotropin FSH; and (3) exposures to FSH followed by Clomiphene. Histological and immunohistochemical results demonstrate that in the lizard Podarcis sicula during the mating period, Clomiphene as well as FSH determines the breakdown of spermatogenesis and the epididymal regression, presumably through estrogens input as indirectly demonstrated by the appearance of ERα and vitellogenin in the liver. The ability of Clomiphene to restore the gonadal natural condition after FSH treatment is also demonstrated. Finally, data indicate that lizard testis and epididymis control their morphophysiology regulating the intracellular presence of ERα

Molecular identification of estrogen receptors (ERα and ERβ) and their differential expression during VTG synthesis in the liver of lizard Podarcis sicula

In non-mammalian vertebrates yolk deposition in the oocytes is a hormone-dependent, gender-specific process. Produced by the ovary under gonadotropin stimulation, Estradiol 17-beta (E(2)) plays a key role in the liver synthesis of vitellogenin (VTG) which in turn is taken up by vitellogenic oocytes in the ovary. In many species a negative role in liver synthesis of VTG in females is also played by progesterone. Experimental administration of E(2) induces the expression of the VTG silent gene also in the liver of males of all the species studied. However, the role of the two isoforms of estrogen receptors, ERalpha and beta, in this process is still unclear. In order to elucidate what kind of ER is involved in the liver synthesis of VTG in the lizard Podarcis sicula, we obtained by means of RT-PCR two fragments of 430bp and 130bp from total ovarian mRNA, encoding respectively for ERalpha and ERbeta. Expression analysis of these two specific isoforms of ERs in the liver showed that in non-breeding females, and in wildlife untreated males only ERbeta is expressed. In breeding vitellogenic females and in E(2)-treated males both alpha and beta receptors are expressed. Furthermore, in females experimentally treated with progesterone during the breeding period, expression of ERalpha disappears. Conversely, treatment of females with E(2) in the non-breeding period induces expression of ERalpha. Immunohistochemical analysis and Western blotting showed that the presence of irVTG in liver and plasma is always parallel to hepatic expression of ERalpha in all the different experimental conditions. Our data strongly suggest that expression of ERalpha may be necessary for VTG synthesis in Podarcis. The possible modulatory role of ERbeta is also discussed

Gene expression profile of Estrogen Receptors alpha and beta in rat brain during aging and following high fat diet.

The “sex-hormone” estrogen-17β promotes several cognitive functions and is a master regulator of brain bioenergetics via the estrogen receptors α and β (ERα and ERβ). In this work, by using Real Time PCR analysis, we evaluated the effect of aging and high fat diet (HFD) on ERα and ERβ expression in rat hippocampus and cortex. In young rats ERβ is abundant in cortex and ERα in hippocampus. During the aging, in the cortex we observe a general decrease in ERα and ERβ expression; in hippocampus ERα increases and ERβ decreases. ERs expression patterns in rat brain are also affected by the administration of a HFD. In cortex, after 4 weeks of HFD ERβ transcripts are down-regulate whereas ERα levels remain unchanged; after 12 weeks both ERα and ERβ expression is up-regulated. In hippocampus, the level of ERβ transcripts does not change following HDF, whereas ERα expression is affected by HDF, in a time-dependent manner: it increases after the 4 weeks treatment and decreases after 12 weeks. Possible involvements of these receptors in the control of cortex and hippocampus functions during aging and in the modulation of energetic metabolism and feeding behaviour are discussed

How Glyphosate Impairs Liver Condition in the Field Lizard Podarcis siculus (Rafinesque-Schmaltz, 1810): Histological and Molecular Evidence

The potential toxicity of glyphosate, a widely used broad-spectrum herbicide, is currently a great matter of debate. As vertebrate insectivores, lizards protect plants from herbivorous insects increasing plant biomass via the trophic cascade and represent an important link between invertebrates and higher predators. A negative effect of glyphosate on lizards’ survival could have major impacts at the ecological levels. In this study, we investigated the effects of the exposure to low doses of glyphosate on the liver of the wall lizard Podarcis siculus, a suitable bioindicator of soil pollution. Two different doses of pure glyphosate (0.05 and 0.5 μg/kg body weight) were orally administered every other day for 3 weeks to sexually mature males and females. The results demonstrated that both doses, despite being very low, are toxic for the liver that showed clear signs of suffering, regardless of sex. The histological analysis provided a scenario of severe hepatic condition, which degenerated until the appearance of fibrotic formations. The morphological observations were consistent with a loss of liver physiological functions. Immunocytochemical investigations allowed us to detect an involvement of antioxidant/cytoprotective proteins, such as superoxide dismutase 1 (Cu/Zn SOD, known as SOD1), glutathione peroxidase 1 (GPx1), metallothionein (MT), and tumor suppressor protein 53, (p53) suggesting that the liver was trying to react against stress signals and damage induced by glyphosate. Finally, in situ hybridization and Real-Time PCR analysis showed the upregulation of estrogen receptor α and vitellogenin gene expression, thus demonstrating the xenoestrogenic action of glyphosate. The imbalance of the hormonal homeostasis could threaten the lizards’ reproductive fitness and survival, altering the trophic cascade

Estrogen-dependent, extrahepatic synthesis of vitellogenin in male vertebrates: A mini-review.

In the last years, the hormonal balance is threatened by the interferences of substances with hormone-like action (endocrine disruptor chemicals, EDCs) that may harm animal reproduction. Most EDCs are resistant to environmental degradation and are considered ubiquitous contaminants. EDCs may have synthetic or natural origins. Pesticides used in intensive agriculture contain large amounts of chemicals with estrogenic properties, such as the alkylphenol nonylphenol (NP). Besides, animal feeding operations are important sources of natural estrogen metabolites introduced into the environment through manure application in organic farming. In both cases, EDCs can reach animals, including humans particularly at risk due to their position in the food chain. This is the reason for which it is important to use terrestrial vertebrates as sentinels in soil biomonitoring programmes. Today, the most validated biomarker of estrogenic exposure is the expression in male liver of the vitellogenin (VTG), an estrogen-dependent glycolipophosphoprotein naturally expressed only in the liver of oviparous females during the reproductive season. This report summarizes the data available on the EDC-dependent expression and the synthesis of VTG in male vertebrates, highlighting our latest studies that demonstrate the ability of testis and epididymis of the lacertid Podarcis sicula to synthesize VTG following estrogenic exposure. These findings provide, for the first time, evidence on an extrahepatic expression and synthesis of VTG in a terrestrial vertebrate and lay the groundwork for a new value of the VTG as a biomarker of environmental contamination. In addition, the results open a new scenario on the role of VTG in cells other than oocytes