Steroid Regulation Research Articles

A Z-Linked E3 Ubiquitin Ligase Cs-rchy1 Is Involved in Gametogenesis in Chinese Tongue Sole, Cynoglossus semilaevis.

Simple SummaryThe sexual growth dimorphism prevails in animals and this phenomenon is even more obvious in marine fish, so understanding the mechanism of gonadal development and gametogenesis is of great importance for sex control, thus increased productivity in aquaculture. In mammal, ubiquitin ligase plays a versatile role in gonadal development and spermatogenesis, whereas its function in fish is little reported. Using Cynoglossus semilaevis (one-year-old female individual usually grows 2–4 times bigger than male) as the fish model, a Z-chromosome linked ubiquitin ligase neurl3 was previously identified and characterized, which suggested its involvement in spermatogenesis. However, in this study, characterization of another Z-chromosome linked ubiquitin ligase Cs-rchy1 suggested it might function both in spermatogenesis and oogenesis, as well as the potential role in growth. These data may provide the genetic resource for gene editing or marker exploration in future.Ubiquitin ligase (E3) plays a versatile role in gonadal development and spermatogenesis in mammals, while its function in fish is little reported. In this study, a Z-chromosome linked ubiquitin ligase rchy1 in C. semilaevis (Cs-rchy1) was cloned and characterized. The full-length cDNA was composed of 1962 bp, including 551 bp 5′UTR, 736 bp 3′UTR, and 675 bp ORF encoding a 224-amino-acid (aa) protein. Cs-rchy1 was examined among seven different tissues and found to be predominantly expressed in gonads. In testis, Cs-rchy1 could be detected from 40 days post hatching (dph) until 3 years post hatching (yph), but there was a significant increase at 6 months post hatching (mph). In comparison, the expression levels in ovary were rather stable among different developmental stages. In situ hybridization showed that Cs-rchy1 was mainly localized in germ cells, that is, spermatid and spermatozoa in testis and stage I, II and III oocytes in ovary. In vitro RNA interference found that Cs-rchy1 knockdown resulted in the decline of sox9 and igf1 in ovarian cell line and down-regulation of cyp19a in the testicular cell line. These data suggested that Cs-rchy1 might participate in gonadal differentiation and gametogenesis, via regulating steroid hormone synthesis.

Cereal Vinegar Sediment Alleviates Spontaneous Ulcerative Colitis in Il-10 Deficient Mice.

Cereal vinegar sediment (CVS) is precipitation generated during the preservation of vinegar. It has various functions such as anti-inflammatory, anti-tumor, hypoglycemic, and hypolipidemic. This study evaluates the effects of CVS on spontaneous colitis in Il-10-/- mice. CVS (1 g kg-1 body weight) is administered to mice for 42 days. CVS alleviated epithelium damage, inhibited myeloperoxidase (MPO) activity and malondialdehyde (MDA) level, decreased gene expression of tumor necrosis factor (Tnf )-a, inducible nitric oxide synthase (Inos), Interleukin(Il-23) in colon tissues is found. CVS also inhibited secretion of IL-2, IL-6, IL-13, Granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GMCSF), Interferon (IFN)-γ, and Regulated upon Activation, Normal T Cell Expressed and Presumably Secreted (RANTES) in serum. While CVS enhanced Regenerating Family Member 3 Gamma (Reg3γ), Mucin (Muc2, Muc3, and Muc4 gene expression, promoted intestinal epithelial cells to secrete Muc-2, and increased the content of acetic acid in intestinal tract of Il-10-/- mice. Additionally, CVS altered the composition of the gut microbiota by promoting the abundance of Akkermansia, Alistipes, and Lactobacillus, while inhibiting Desulfovibrio and Clostridium sensu stricto 1. These changes may be related to the regulation of steroid, fatty acids, and bile acid biosynthesis. This study demonstrated that CVS ameliorates spontaneous ulcerative colitis in Il-10-/- mice, which suggests CVS supplementation may serve as a protective dietary nutrient against colitis.

The Therapeutic Intervention of Sex Steroid Hormones for Sarcopenia

Sarcopenia, characterized by the excessive loss of skeletal muscle mass, strength, and function, is associated with the overall poor muscle performance status of the elderly, and occurs more frequently in those with chronic diseases. The causes of sarcopenia are multifactorial due to the inherent relationship between muscles and molecular mechanisms, such as mitochondrial function, inflammatory pathways, and circulating hormones. Age-related changes in sex steroid hormone concentrations, including testosterone, estrogen, progesterone, and their precursors and derivatives, are an important aspect of the pathogenesis of sarcopenia. In this review, we provide an understanding of the treatment of sarcopenia through the regulation of sex steroid hormones. The potential benefits and future research emphasis of each sex steroid hormone therapeutic intervention (testosterone, SARMs, estrogen, SERMs, DHEA, and progesterone) for sarcopenia are discussed. Enhanced understanding of the role of sex steroid hormones in the treatment for sarcopenia could lead to the development of hormone therapeutic approaches in combination with specific exercise and nutrition regimens.

Alpha/Beta Hydrolase Domain-Containing Protein 2 Regulates the Rhythm of Follicular Maturation and Estrous Stages of the Female Reproductive Cycle.

Mammalian female fertility is defined by a successful and strictly periodic ovarian cycle, which is under the control of gonadotropins and steroid hormones, particularly progesterone and estrogen. The latter two are produced by the ovaries that are engaged in controlled follicular growth, maturation, and release of the eggs, i.e., ovulation. The steroid hormones regulate ovarian cycles via genomic signaling, by altering gene transcription and protein synthesis. However, despite this well-studied mechanism, steroid hormones can also signal via direct, non-genomic action, by binding to their membrane receptors. Here we show, that the recently discovered membrane progesterone receptor α/β hydrolase domain-containing protein 2 (ABHD2) is highly expressed in mammalian ovaries where the protein plays a novel regulatory role in follicle maturation and the sexual cycle of females. Ablation of Abhd2 caused a dysregulation of the estrous cycle rhythm with females showing shortened luteal stages while remaining in the estrus stage for a longer time. Interestingly, the ovaries of Abhd2 knockout (KO) females resemble polycystic ovary morphology (PCOM) with a high number of atretic antral follicles that could be rescued with injection of gonadotropins. Such a procedure also allowed Abhd2 KO females to ovulate a significantly increased number of mature and fertile eggs in comparison with their wild-type littermates. These results suggest a novel regulatory role of ABHD2 as an important factor in non-genomic steroid regulation of the female reproductive cycle.

MP33-11 OBESITY-ASSOCIATED PROINFLAMMATORY CYTOKINES REGULATE PROSTATE CANCER CELL PROLIFERATION AND MIGRATION

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I (MP33)1 Sep 2021MP33-11 OBESITY-ASSOCIATED PROINFLAMMATORY CYTOKINES REGULATE PROSTATE CANCER CELL PROLIFERATION AND MIGRATION Xiaobo Wu, Christina Sharkey, Zongwei Wang, and Aria Olumi Xiaobo WuXiaobo Wu More articles by this author , Christina SharkeyChristina Sharkey More articles by this author , Zongwei WangZongwei Wang More articles by this author , and Aria OlumiAria Olumi More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002042.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Previous studies revealed that obesity is significantly associated with a higher incidence of prostate cancer (PCa). Meanwhile, obesity-associated inflammation plays an important role in tumorigenesis. Here we wished to identify the downstream signaling pathways of obesity-associated proinflammatory cytokines IL-6 and TNF-α’s effects on prostate cancer cell proliferation and migration. METHODS: Adipocytes and THP-1 macrophages were differentiated and treated with myristic acid (MA) to induce inflammatory cytokines, and conditioned media (CM) were collected to treat prostate epithelial cancer, androgen-dependent LNCaP and androgen-independent PC-3 cell lines. Cells were also treated with interleukin-6 (IL-6) 10ng/ml, tumor necrosis factor-alpha (TNF-α) 10ng/ml, oncometabolite R-2-hydroxyglutarate 50µM (R-2-HG) and Dutasteride 10µM (SRD5A1 inhibitor). Prostate cancer cell migration and proliferation were assessed by scratch assay (0h, 24h, 48h) and MTT assay (0h, 24h, 48h, 72h) and analyzed by 2-way ANOVA. Enzymes regulating steroid hormone metabolism SRD5A1, AKR1C1, AKR1C3, HSD3B2, CYP17A1 and UGT2B15 were determined by immunocytochemistry and qPCR. The protein level of SRD5A1 was measured by western blot. RESULTS: The proliferation of LNCaP and PC-3 cells was significantly stimulated by adding of CM for 24h, 48h, and 72h. The migration of PC-3 cells was also promoted by CM. Addition of testosterone did not change the effect of CM. Administration of IL-6, TNF-α and R-2-HG significantly increased the proliferation and migration of LNCaP and PC-3 cells. IL-6 also increased the expression of SRD5A1.. The expression of AKR1C1 and HSD3B2 was downregulated, and UGT2B15 was upregulated by TNF-α. SRD5A1 was upregulated either by R-2-HG alone or combining with IL-6 and TNF-α, but downregulated by Dutasteride alone. CONCLUSIONS: Our study suggests that obesity-associated proinflammatory cytokines, IL-6 and TNF-α, affect prostate cancer cells’ proliferation and migration in an androgen-independent manner. Synergistic targeting oncometabolite R-2-HG and proinflammatory cytokines may improve the management of androgen-independent prostate cancer. Source of Funding: NIH/NIDDK © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e610-e611 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Xiaobo Wu More articles by this author Christina Sharkey More articles by this author Zongwei Wang More articles by this author Aria Olumi More articles by this author Expand All Advertisement Loading ...

The influence of dietary arachidonic acid on growth, fatty acid profile and sex steroid hormones of F2 generation Chinese sturgeon (Acipenser sinensis)

An 22-week feeding trial was performed to evaluate the effects of dietary arachidonic acid (ARA) on the growth performance, tissue fatty acid, ARA metabolites, sex steroid hormones and steroidogenesis-related gene expression in F2 generation Chinese sturgeon. Four diets with different ARA levels (0.0 %, 0.5 %, 1.0 % and 2.0 % in the diets) were formulated. The results showed that dietary ARA did not affect final body weight (FBW), weight gain rate (WGR), feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI), viscerosomatic index (VSI), condition factor (CF) and gonadalsomatic index (GSI) (P > 0.05), while tissue ARA deposition was significantly effected by the dietary ARA (P < 0.05). In the ovary and liver, the metabolites of prostaglandin E2 (PGE2) and 5-hydroxyeicosatetraenoic acid (5-HETE) increased significantly with increasing dietary ARA (P < 0.05). In the muscle, the contents of the two metabolites in the 1.0 % ARA diet were significantly higher than that the 0.0 % and 2.0 % ARA diets (P < 0.05). Significantly decreased liver and ovary estradiol (E2), testosterone (T) levels were found in the 0.0 % ARA diet compared with 2.0 % ARA diet (P < 0.05). In the ovary and liver, the contents of vitellogen (Vtg) increased significantly with increasing dietary ARA (P < 0.05). E2 and Vtg contents in the serum of fish fed the 1.0 % ARA diet were significantly higher than the 0.0 % ARA diet (P < 0.05). These results indicated that dietary ARA did not affect growth, but ARA levels greater than 1 % in the diets plays vital roles in the ARA deposition, metabolism, steroid regulation and Vtg synthesis in F2 generation female Chinese sturgeon. The best efficiency was achieved 1 % ARA, based on roles or cost effective, which were beneficial to promote ovary development.

Genetic comprehension of organophosphate flame retardants, an emerging threat to prostate cancer

In recent years, organophosphate ester flame retardants (OPFRs), which have been regarded as alternatives for brominated flame retardants (BFRs), have become widely used in building materials, textiles, and electric equipment. Elucidating the relationship between OPFRs and tumors holds great significance for the treatment and prevention of diseases. In this work, we found a new method for predicting the correlation between the interactive genes of OPFRs and tumors. Transcriptome profiles and OPFR information were obtained from The Cancer Genome Atlas and the Genotype–Tissue Expression, Comparative Toxicogenomics, and PharmMapper databases. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis showed that interactive genes were mainly enriched in prostate cancer, steroid metabolic process, and steroid hormone regulation. Furthermore, protein–protein interaction network analysis revealed 33 biological hub genes. The operating characteristic curves and survival analysis showed the role of key genes in predicting the prognosis of prostate cancer. Gene target prediction and gene set variation analysis proved that OPFRs and their metabolites exert potential effects on prostate cancer. Colony formation assay showed that the cells with AR, mTOR and DDIT3 knockdown could remarkably mitigate the cell proliferation ability in both PC-3 and LNCap cells. Transwell assay demonstrated that the silencing of AR, mTOR and DDIT3 could significantly inhibit the cell invasion capacity of prostate cells. Triphenyl phosphate (TPP) significantly increase the cell proliferation ability and promote cell invasion capacity. AR, mTOR and DDIT3 in the PC-3 and LNCap cells were significantly upregulated with 10–6 M TPP treated.

The Effect of Chemotherapy in Gestational Trophoblastic Disease on Infertility: A Review on Pathophysiologic Mechanism

Today, young women infertility through chemotherapy has become a global challenge. Chemotherapy destructs the malignant cells by reactive oxygen species (ROS) production and inflammatory factors secretion; these factors can also destruct the ovarian and uterine cells. Infertility usually happens as a result of ovarian and uterine cells apoptosis, as well as dysfunction in these organs. Signaling pathways activated by chemotherapy lead to increased activation of follicles and depletion of the follicular pool. In addition, excessive secretion of sex hormones leads to follicles activation and infertility in women. Mesenchymal stem cells (MSC) use have been reported to be a great help in restoring the function of ovarian and uterine cells. On the other hand, they can regulate sex hormone secretion. Finally, the use of MSCs as a suitable treatment strategy can help restore the function of reproductive cells and treat infertility. *Corresponding Author: Maryam Ebrahimi; ORCID iD: 0000 0001 9685 7882; Email: drmkhoddami@yahoo.com Please cite this article as: Eslamnik P, Moghaddasi M, Malmir A, Sadeghi-Goughari B, Javan MR, Ebrahimi M. The Effect of Chemotherapy in Gestational Trophoblastic Disease on Infertility: A Review on Pathophysiologic Mechanism. Arch Med Lab Sci. 2021;7:1-8 (e8). https://doi.org/10.22037/amls.v7.34369

A Potential Role and Contribution of Androgens in Placental Development and Pregnancy.

Successful pregnancy requires the establishment of a highly regulated maternal–fetal environment. This is achieved through the harmonious regulation of steroid hormones, which modulate both maternal and fetal physiology, and are critical for pregnancy maintenance. Defects in steroidogenesis and steroid signaling can lead to pregnancy disorders or even fetal loss. The placenta is a multifunctional, transitory organ which develops at the maternal–fetal interface, and supports fetal development through endocrine signaling, the transport of nutrients and gas exchange. The placenta has the ability to adapt to adverse environments, including hormonal variations, trying to support fetal development. However, if placental function is impaired, or its capacity to adapt is exceeded, fetal development will be compromised. The goal of this review is to explore the relevance of androgens and androgen signaling during pregnancy, specifically in placental development and function. Often considered a mere precursor to placental estrogen synthesis, the placenta in fact secretes androgens throughout pregnancy, and not only contains the androgen steroid nuclear receptor, but also non-genomic membrane receptors for androgens, suggesting a role of androgen signaling in placental function. Moreover, a number of pregnancy disorders, including pre-eclampsia, gestational diabetes, intrauterine growth restriction, and polycystic ovarian syndrome, are associated with abnormal androgen levels and androgen signaling. Understanding the role of androgens in the placenta will provide a greater understanding of the pathophysiology of pregnancy disorders associated with androgen elevation and its consequences.

Chronic low-level perfluorooctane sulfonate (PFOS) exposure promotes testicular steroidogenesis through enhanced histone acetylation

Perfluorooctane sulfonate (PFOS), an artificial perfluorinated compound, has been associated with male reproductive disorders. Histone modifications are important epigenetic mediators; however, the impact of PFOS exposure on testicular steroidogenesis through histone modification regulations remains to be elucidated. In this study, we examined the roles of histone modifications in regulating steroid hormone production in male rats chronically exposed to low-level PFOS. The results indicate that PFOS exposure significantly up-regulated the expressions of StAR, CYP11A1 and 3β-HSD, while CYP17A1 and 17β-HSD were down-regulated, thus contributing to the elevated progesterone and testosterone levels. Furthermore, PFOS significantly increased the histones H3K9me2, H3K9ac and H3K18ac while reduced H3K9me3 in rat testis. It is known that histone modifications are closely involved in gene transcription. Therefore, to investigate the association between histone modifications and steroidogenic gene regulation, the levels of these histone marks were further measured in steroidogenic gene promoter regions by ChIP. It was found that H3K18ac was augmented in Cyp11a1 promoter, and H3K9ac was increased in Hsd3b after PFOS exposure, which is proposed to result in the activation of CYP11A1 and 3β-HSD, respectively. To sum up, chronic low-level PFOS exposure activated key steroidogenic gene expression through enhancing histone acetylation (H3K9ac and H3K18ac), ultimately stimulating steroid hormone biosynthesis in rat testis.

1196-P: Perineuronal Nets in MeA and Arc Are Influenced by Gonadectomy

Background: Recent studies reveal perineuronal nets (PNNs), extracellular matrix specializations that restrict the plasticity of neurons, are influenced by pubertal status. Here we show that PNN are changed by gonadectomy (GDX) both in male and female mice in medial amygdala (MeA) and arcuate nucleus (Arc). Method: 49 wild type mice are divided into 16 groups by sex, gonadectomy, diet, nutritional status (fed or fasted groups). Fat and lean body mass were assessed by QMR at baseline and before perfusion. Body weight, food intake, and blood glucose were measured weekly for five weeks. Images were captured in MeA, Arc, paraventricular hypothalamus (PVH), anterior hypothalamus (AH), and an intense WFA positive radial area near PVH, which we named ‘periPVH’ after labelling of Wisteria floribunda agglutinin (WFA). Results: In MeA region, female mice have significantly lower total fluorescence intensity (TFI) (Estimate, 95% Confidence Interval: –448887.02, –578807.10 – –318966.94; p &amp;lt;0.0001) and total number of PNNs (–319.67, –379.34258 – –259.99075: p&amp;lt;0.0001) than male mice. Notably, in female, OVX mice have lower mean fluorescence intensity (MFI) (Mean ± SD: 14.5 ± 3.3 vs. 19.6 ± 4.1; p = 0.003) and TFI (64907.8 (58406.6–96971.3) vs. 123613.9 (83545.9–149131.5); p = 0.015) than sham mice, while the total number of PNNs (Mean ± SD: 63 ± 23 vs. 55 ± 3; p = 0.391) shows no significant difference. Male castration mice have lower total number of PNNs (382 ± 141 vs. 55 ± 22; p &amp;lt;0.0001) and TFI (439343.5 (366431.4–787461.9) vs. 168416.3 (141273.0–258020.8); p &amp;lt;0.0001) than sham mice, while the MFI shows no significant difference. In Arc, mice experiencing GDX have significantly lower MFI (–3.63, –5.61303 – –1.63967; p &amp;lt;0.001), TFI (–28525.8, –45357.46 – –11694.16; P&amp;lt;0.01) or total number of PNNs (–15.3, –29.24 – –1.41; p =0.036) than mice not. PVH, AH, and periPVH don’t show associations with sex, GDX, diet or nutritional status in our observation. Summary: Our novel discovery indicates the potential role PNN might play in sex hormone regulation in brain. Disclosure N. Zhang: None. Y. Xu: None. Funding National Institutes of Health (R01DK117281)

Genetic and in silico analysis show a role of SMAD3 on recurrent pregnancy loss

Recurrent pregnancy loss (RPL) is one of the most common reproductive failures affecting 1–5% of couples. Smad3 is an effector of signalling of the Transforming Growth Factors-β superfamily (TGF-β), regulating the transcription of several target genes of these cytokines. The objective of this study was to evaluate the influence of a variant on SMAD3 (rs17293443) in RPL. A case-control study was carried out with 149 women who experienced RPL and 159 controls, as well as bioinformatics tools to determine the role of this variant in this condition. Our study showed an allelic (p = 0.023) and genotypic (p < 0.01) association of this variant with the RPL. Our functional in silico predictions suggest that this variant causes a change in SMAD3 expression levels. Alterations in the expression of this gene can directly compromise the Smad3-dependent signalling pathway that is fundamental for key processes for gestation such as steroid hormone regulation and implantation, as demonstrated by ontologies analyses performed and the literature. Our findings regarding the involvement of Smad3 on RPL are a novelty in this field, and they seem to be promising to the clinical management of this condition.

Seasonality, sex-specificity and transcriptional regulation of hepatic leptin system in spotted snakehead Channa punctata

The present study deals with sex-specific reproductive phase-dependent variation and sex steroids-induced transcriptional regulation of hepatic lep and lepr in nutritionally valuable spotted snakehead, Channa punctata. The data on seasonality reveals sex-specific variation in pattern of lep transcription where a high level was recorded during resting and postspawning quiescent phases in female while during resting and spawning phases in male. Unlike lep, lepr exhibited similar expression pattern along the reproductive phases in both the sexes. As compared to female, a three-fold higher expression of lep was detected in male during reproductively active phase only. However, no sexual dimorphism was evidenced in lepr either during active or quiescent phase. To explore the implication of sex steroids in regulation of leptin system, we correlated levels of plasma testosterone (T) and 17β-estradiol (E2) with leptin system in males as well as females. Further, criss-cross in vivo and in vitro experiments with dihydrotestosterone (DHT) and E2 were conducted in male and female spotted snakehead. The leptin system was downregulated after DHT administration in both the sexes. However, with E2, a marked decrease was evidenced in male only. The sex-wise variable response of leptin system to sex steroids was validated by in vitro experiments wherein liver fragments from male and female fish were incubated individually with both the sex steroids. In conclusion, sex steroids modulate hepatic leptin system differentially depending on sex and reproductive state of spotted snakehead.

Harderian SOX9: Molecular characterization and its dimorphic expression in hamster

The molecular action of SOX9 can promote lipogenesis. Because the hamster Harderian gland (HG) synthesizes lipids and exhibits sexual dimorphism, this study aimed to identify and characterize Harderian SOX9. We examined the tissue distribution and expression profiles of SOX9 in hamster Mesocricetus auratus HGs. The full-length SOX9 cDNA sequence [3649-base pairs (bp)] contains an 81-bp 5′ untranslated region (UTR), a 3′ UTR of 2044-bp, an open reading frame (ORF) of 1524-bp, and a polyadenylation signal (AATAAA) at 19-bp upstream of poly(A) tail. The cDNA encodes a 507 amino acid protein containing the potential DNA-binding domain known as the HMG box. BLAST analysis revealed 99%, 99%, and 97% identity with the SOX9 of mouse, rat, and human, respectively. High expression levels were also observed in the testis, cerebellum, and hypothalamus. qPCR analysis demonstrated that SOX9 is expressed more abundantly in the HGs of males than in females. Sexually dimorphic expression of SOX9 suggests that differential expression between male and female HGs could be under the regulation of sex steroids. SOX9 might play a similar role in regulating exocrine secretions of lipids; these could occur downstream of FGF signaling – as found during embryogenesis – and/or androgen signaling.

Testosterone Treatment Mimics Seasonal Downregulation of Dopamine Innervation in the Auditory System of Female Midshipman Fish

In seasonally breeding vertebrates, hormones coordinate changes in nervous system structure and function to facilitate reproductive readiness and success. Steroid hormones often exert their effects indirectly via regulation of neuromodulators, which in turn can coordinate the modulation of sensory input with appropriate motor output. Female plainfin midshipman fish (Porichthys notatus) undergo increased peripheral auditory sensitivity in time for the summer breeding season, improving their ability to detect mates, which is regulated by steroid hormones. Reproductive females also show differences in catecholaminergic innervation of auditory circuitry compared with winter, non-reproductive females as measured by tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholaminergic synthesis. Importantly, catecholaminergic input to the inner ear from a dopaminergic-specific forebrain nucleus is decreased in the summer and dopamine inhibits the sensitivity of the inner ear, suggesting that gonadal steroids may alter auditory sensitivity by regulating dopamine innervation. In this study, we gonadectomized non-reproductive females, implanted them with estradiol (E2) or testosterone (T), and measured TH immunoreactive (TH-ir) fibers in auditory nuclei where catecholaminergic innervation was previously shown to be seasonally plastic. We found that treatment with T, but not E2, reduced TH-ir innervation in the auditory hindbrain. T-treatment also reduced TH-ir fibers in the forebrain dopaminergic cell group that projects to the inner ear, and likely to the auditory hindbrain. Higher T plasma in the treatment group was correlated with reduced-ir TH terminals in the inner ear. These T-treatment induced changes in TH-ir fibers mimic the seasonal downregulation of dopamine in the midshipman inner ear and provide evidence that steroid hormone regulation of peripheral auditory sensitivity is mediated, in part, by dopamine.

Cardiac Arrest in a Child with Non-classic Lipoid Congenital Adrenal Hyperplasia Associated with a New STAR Gene Mutation

Abstract Introduction: Steroidogenic acute regulatory (STAR) protein regulates steroid hormone synthesis by transporting cholesterol into mitochondria. STAR gene mutations lead to lipoid congenital adrenal hyperplasia (LCAH), the rare but most severe form of congenital adrenal hyperplasia in children. We present an unusual case with an episode of cardiac arrest in a young girl during an acute febrile illness and later she was diagnosed with adrenal insufficiency secondary to a non-classic LCAH. Case: 2-year 11-month-old previously healthy white female was brought to an urgent care clinic due to severe lethargy and a seizure-like activity during a fever illness. She was found to have an undetectable blood glucose level and went into cardiac arrest shortly after arrival. CPR was performed for approximately 11 minutes. She then developed sever respiratory distress and was intubated. She was transferred to the PICU with IV sodium bicarbonate given en route. On admission, her body weight was 13.26 kg (36.80th percentile), height 90 cm (17.56th percentile), and BMI 16.17 (62.88th percentile). Her physical exam revealed normal external female genitalia and normal skin pigmentation. Lab evaluation revealed normal sodium and potassium but elevated anion gap, hyperuricemia, elevated creatinine kinase, abnormal liver function tests and abnormal coagulation profile. Brain MRI revealed findings consistent with hypoxic-ischemic encephalopathy. Renal function improved within 24 hours and hepatic function returned to normal after 20 days. Due to her severe hypoglycemic event, a high-dose ACTH stimulation test was performed. The results were consistent with adrenal insufficiency: baseline cortisol level, 7.3 μg/dL; 30 minutes cortisol, 7.8 μg/dL; 60 minutes cortisol, 9 μg/dL (normal response, ≥18 mcg/mL at 30 or 60 minutes). The baseline ACTH level was significantly elevated, 1688 pg/mL (0–46) as well as the renin activity, 24.3 ng/hour (1.7–11.2). Genetic testing revealed a 46 XX karyotype. STAR gene analysis identified compound heterozygosity; a novel deletion (c.811delC, p.Leu271Cysfs*50) and a previously reported missense mutation (c.661G&amp;gt;A, p.Gly221Ser). The girl is now 11 years old and exhibits normal growth, normal cognitive development, and she has developed early signs of puberty (Tanner stage 2 for breast). She takes daily hydrocortisone, fludrocortisone and stress dose hydrocortisone as needed. Conclusion: In non-classic LACH, the onset is generally late or not acute. Initial clinical features are variable and nonspecific. For this reason, non-classic LCAH may be overlooked. Adrenal crisis is a life-threatening complication, and it is important that clinicians are aware of the clinical features of non-classic LCAH and consider it in the differential diagnoses. Genetic testing for STAR should be considered in individuals with non-autoimmune primary adrenocortical insufficiency.

Development of a Novel Tetracycline-Inducible Kiss1-Cre Mouse Line for Temporally Controlled Gene Deletion in Kisspeptin Neurons

The tetracycline (Tet)-controlled inducible system is the most widely used reversible system for transgenic expression in mice. Previously, we generated a GnRH-CreTeR mouse model, using a first-generation Tet-inducible system to temporally induce expression of Cre recombinase in GnRH neurons. Recently, the Tet-inducible system has undergone several modifications to significantly reduce previous limitations that include leaky background expression and lower sensitivity to tetracycline induction. Therefore, we have developed a novel mouse model bearing a Tet-inducible kisspeptin-Cre allele (iKiss-Cre mouse) that will enable temporal control over the selective deletion of genes from Kiss1 neurons. This temporally controlled gene deletion will eliminate a longstanding technical limitation of conventional steroid receptor knockout models in which steroid regulation of the axis is confounded by steroid developmental and organizational effects in the reproductive axis. Two mouse lines were generated. The first line targets kisspeptin neurons with a third generation Tet-inducible reverse tetracycline transactivator (rtTA, Tet-On 3G) expressed under the control of the Kiss1 allele. Using CRISPR-Cas9 technology, we inserted a cassette containing an internal ribosome entry site (IRES) sequence followed by the rtTA downstream of the Kiss1 coding region as was previously done using Cre recombinase. Transcription of the recombinant Kiss1 allele yields a bicistronic messenger RNA, from which both kisspeptin and rtTA are independently translated. The second mouse line, TRE-Cre mice, was constructed to express Cre recombinase under control of the PTRE3G promoter. The PTRE3G promoter is bidirectional to allow simultaneous monitoring of Cre expression and a second florescent protein reporter (ZsGreen1). The Kiss1-rtTA only binds to and activates the PTRE3G promoter in the presence of doxycycline (an analog of tetracycline). Offspring from breeding of these mouse lines, iKiss-Cre, result in a system capable of generating Tet-induced expression of Cre recombinase in kisspeptin neurons. In order to document specificity and sensitivity of this system, we performed immunofluorescent staining and observed colocalization of kisspeptin with Cre recombinase in brain sections of iKiss-Cre mice only after doxycycline treatment. We will use this model to investigate negative feedback actions of E2 on the adultpulse generator after E2 exerts its organizational actions on maturing kisspeptin neurons. We expect that this mouse model will become a major tool used by the neuroendocrine community and serve as proof of principle for development of similar inducible knockout models employing the current generation of inducible methodologies.

Investigation on Sex Hormone-Disruption Effects of Two Novel Brominated Flame Retardants (DBDPE and BTBPE) in Male Zebrafish (Danio rerio) and Two Human Cell Lines (H295R and MVLN)

Decabromodiphenyl ethane (DBDPE) and 1,2-bis(2,4,6-tribromo-phenoxy) ethane (BTBPE) are novel brominated flame retardants (NBFRs) and have been detected in variety of environment and biota. Although sex endocrine-disrupting potential has been suggested in experimental studies, their adverse effects on sex steroid hormones and underlying mechanisms are largely unclear. The purpose of the present study is to investigate the sex hormone-disrupting effects of two NBFRs using in vivo and in vitro models together. For this, male zebrafish (Danio rerio) along with human adrenocortical carcinoma (H295R) and breast carcinoma (MVLN) cell lines were employed. In male zebrafish, 14-day exposure to DBDPE significantly increased 17β-estradiol (E2) concentrations. Disruption of sex hormone regulation was also suggested after exposure to BTBPE, i.e., the increasing trend of E2 levels, E2/11-ketotestosterone (11-KT) ratio, and estrogen receptor-alpha (erα) and erβ gene expression levels. In H295R cells, an E2/T ratio showed an increasing trend by DBDPE exposure, but transcriptions of major genes in steroidogenesis pathway were not affected. Taken together, our observation implies that two NBFRs could cause the sex hormone disruption potential in male zebrafish and H295R cells but probably not through alteration of steroidogenesis pathway.

Migratory state and patterns of steroid hormone regulation in the pectoralis muscle of a nomadic migrant, the pine siskin (Spinus pinus)

The endocrine system is known to mediate responses to environmental change and transitions between different life stages (e.g., a non-breeding to a breeding life stage). Previous works from the field of environmental endocrinology have primarily focused on changes in circulating hormones, but a comprehensive understanding of endocrine signaling pathways requires studying changes in additional endocrine components (e.g., receptor densities) in a diversity of contexts and life stages. Migratory birds, for instance, can exhibit dramatic changes in their physiology and behavior, and both sex steroids as well as glucocorticoids are proposed mediators of the transition into a migratory state. However, the role of changes in endocrine signaling components within integral target tissues, such as flight muscles, in modulating the transition into a migratory state remains poorly understood. Here, we examined changes in gene expression levels of and correlational patterns (i.e., integration) between 8 endocrine signaling components associated with either glucocorticoids or sex steroid signaling in the pectoralis muscles of a nomadic migratory bird, the pine siskin (Spinus pinus). The pectoralis muscle is essential to migratory flight and undergoes conspicuous changes in preparation for migration, including hypertrophy. We focus on endocrine receptors and enzymes (e.g., 5α-reductase) that modulate the signaling capacity of circulating hormones within target tissues and may influence either catabolic or anabolic functioning within the pectoralis. Endocrine signaling components were compared between captive birds sampled prior to the expression of vernal migratory preparation and during the expression of a vernal migratory state. While birds exhibited differences in the size and color of the flight muscle and behavioral shifts indicative of a migratory state (i.e., zugunruhe), none of the measured endocrine components differed before and after the transition into the migratory state. Patterns of integration amongst all genes did, however, differ between the two life stages, suggesting the contrasting demands of different life stages may shape entire endocrine signaling networks within target tissues rather than individual components. Our work aligns with previous endocrine studies on pine siskins and, viewed together, suggest additional studies are needed to understand the endocrine system’s role in mediating the development and progression of the vernal migratory state in this species. Further, the patterns observed in pine siskins, a nomadic migrant, differ from previous studies on obligate migrants and suggest that different mechanisms or interactions between endocrine signaling components may mediate the migratory transition in nomadic migrants.

Sex hormones regulate lipid metabolism in adult Sertoli cells: A genome-wide study of estrogen and androgen receptor binding sites

Optimal functioning of Sertoli cells is crucial for spermatogenesis which is under tight regulation of sex hormones, estrogen and androgen. Adult rat Sertoli cells expresses estrogen receptor beta (ERβ) and androgen receptor (AR), both of which regulate gene transcription by binding to the DNA. The present study is aimed to acquire a genome-wide map of estrogen- and androgen-regulated genes in adult Sertoli cells. ChIP-Seq was performed for ERβ and AR in Sertoli cells under physiological conditions. 30,859 peaks in ERβ and 9,594 peaks in AR were identified with a fold enrichment >2 fold. Pathway analysis for the genes revealed metabolic pathways to be significantly enriched. Since Sertoli cells have supportive functions and provide energy substrates to germ cells during spermatogenesis, significantly enriched metabolic pathways were explored further. Peaks of the genes involved in lipid metabolism, like fatty acid, glyceride, leucine, and sphingosine metabolism were validated. Motif analysis confirmed the presence of estrogen- and androgen-response elements (EREs and AREs). Moreover, transcript levels of enzymes involved in the lipid metabolic pathways were significantly altered in cultured Sertoli cells treated with estrogen and androgen receptor agonists, demonstrating functional significance of these binding sites. This study elucidates a mechanism by which sex hormones regulate lipid metabolism in Sertoli cells by transcriptionally controlling the expression of these genes, thereby shedding light on the roles of these hormones in male fertility.