Steroid Acute Regulatory Protein Research Articles

Female baboon adrenal zona fasciculata and zona reticularis regulatory and functional proteins decrease across the life course.

Debate exists on life-course adrenocortical zonal function trajectories. Rapid, phasic blood steroid concentration changes, such as circadian rhythms and acute stress responses, complicate quantification. To avoid pitfalls and account for life-stage changes in adrenocortical activity indices, we quantified zonae fasciculata (ZF) and reticularis (ZR) across the life-course, by immunohistochemistry of key regulatory and functional proteins. In 28 female baboon adrenals (7.5-22.1years), we quantified 12 key proteins involved in cell metabolism, division, proliferation, steroidogenesis (including steroid acute regulatory protein, StAR), oxidative stress, and glucocorticoid and mitochondrial function. Life-course abundance of ten ZF proteins decreased with age. Cell cycle inhibitor and oxidative stress markers increased. Seven of the 12 proteins changed in the same direction for ZR and ZF. Importantly, ZF StAR decreased, while ZR StAR was unchanged. Findings indicate ZF function decreased, and less markedly ZR function, with age. Causes and aging consequences of these changes remain to be determined.

Fluctuating and Stable High Temperatures Differentially Affect Reproductive Endocrinology of Female Pupfish.

For many fishes, reproductive function is thermally constrained such that exposure to temperatures above some upper threshold has detrimental effects on gametic development and maturation, spawning frequency, and mating behavior. Such impairment of reproductive performance at elevated temperatures involves changes to hypothalamic-pituitary-gonadal (HPG) axis signaling and diminished gonadal steroidogenesis. However, how HPG pathways respond to consistently high versus temporally elevated temperatures is not clear. Here, sexually mature Amargosa River Pupfish (Cyprinodon nevadensis amargosae) were maintained under thermal regimes of either stable ∼25°C (low temperature), diurnal cycling temperatures between ∼27 and 35°C (fluctuating temperature), or stable ∼35°C (high temperature) conditions for 50 days to examine effects of these conditions on HPG endocrine signaling components in the pituitary gland and gonad, ovarian and testicular gametogenesis status, and liver gene expression relating to oogenesis. Female pupfish maintained under stable high and fluctuating temperature treatments showed reduced gonadosomatic index values as well as a lower proportion of oocytes in the lipid droplet and vitellogenic stages. Females in both fluctuating and stable 35°C conditions exhibited reduced ovarian mRNAs for steroid acute regulatory protein (star), cholesterol side chain-cleavage enzyme, P450scc (cyp11a1), and 3β-hydroxysteroid dehydrogenase (3bhsd), while ovarian transcripts encoding 11β-hydroxysteroid dehydrogenase (11bhsd) and sex hormone-binding globulin (shbg) were elevated in females at constant 35°C only. Ovarian aromatase (cyp19a1a) mRNA levels were unaffected, but circulating 17β-estradiol (E2) was lower in females at 35°C compared to the fluctuating temperature condition. In the liver, mRNA levels for choriogenins and vitellogenin were downregulated in both the fluctuating and 35°C conditions, while hepatic estrogen receptor 2a (esr2a) and shbg mRNAs were elevated in 35°C females. Taken together, these results demonstrate the potential for elevated temperatures to impair ovarian steroidogenesis and reduce egg envelope and vitellogenin protein production in female C. n. amargosae pupfish, while also shedding light on how thermal regimes that only intermittently reach the upper thermal range for reproduction have differential impacts on reproductive endocrine pathways than constantly warm conditions.

Effect of quercetin on granulosa cells development from hierarchical follicles in chicken

ABSTRACT 1. The bioflavonoid quercetin is a biologically active component, but its functional regulation of granulosa cells (GCs) during chicken follicular development is little studied. To investigate the effect of quercetin on follicular development in laying hens, an in vitro study was conducted on granulosa cells from hierarchical follicles treated with quercetin. 2. The effect of quercetin on cell activity, proliferation and apoptosis of granulosa cells was detected by CCK-8, EdU and apoptosis assays. The effect on progesterone secretion from granulosa cells was investigated by enzyme-linked immunosorbent assay (ELISA). Expression of proliferating cell nuclear antigen (PCNA) mRNA and oestrogen receptors (ERs), as well as the expression of steroid acute regulatory protein (StAR), cytochrome P450 cholesterol side chain cleavage enzyme (P450scc) and 3β-hydroxysteroid dehydrogenase (3β-HSD) mRNA during progesterone synthesis, were measured by real-time quantitative polymerase chain reaction (RT-qPCR). PCNA, StAR and CYP11A1 protein expression levels were detected using Western blotting (WB). 3. The results showed that treatment with quercetin in granulosa cells significantly enhanced cell vitality and proliferation, reduced apoptosis and promoted the expression of gene and protein levels of PCNA. The levels of progesterone secretion increased significantly following quercetin treatment, as did the expression levels of StAR and CYP11A1 using the Western Blot (WB) method. 4. The mRNA expression levels of ERα were significantly upregulated in the 100 ng/ml and 1000 ng/ml quercetin-treated groups, while there was no significant difference in expression levels of ERβ mRNA.

Accustomed to the heat: Temperature and thyroid hormone influences on oogenesis and gonadal steroidogenesis pathways vary among populations of Amargosa pupfish (Cyprinodon nevadensis amargosae)

Many fish experience diminished reproductive performance under atypically high or prolonged elevations of temperature. Such high temperature inhibition of reproduction comes about in part from altered stimulation of gametogenesis by the hypothalamic-pituitary-gonadal (HPG) endocrine axis. Elevated temperatures have also been shown to affect thyroid hormone (TH) signaling, and altered TH status under high temperatures may impact gametogenesis via crosstalk with HPG axis pathways. Here, we examined effects of temperature and 3′-triiodo-L-thyronine (T3) on pathways for gonadal steroidogenesis and gametogenesis in Amargosa pupfish (Cyprinodon nevadensis amargosae) from two allopatric populations: 1) the Amargosa River – a highly variable temperature habitat, and 2) Tecopa Bore – an invariably warm groundwater-fed marsh. These populations were previously shown to differ in TH signaling profiles both in the wild and under common laboratory conditions. Sexually-mature pupfish from each population were maintained at 24 °C or 34 °C for 88 days, after which a subset of fish was treated with T3 for 18–24 h. In both populations, mRNA abundances for follicle-stimulating hormone receptor and luteinizing hormone receptor were higher in the ovary and testis at 24 °C compared to 34 °C. Females from Tecopa Bore – but not from the Amargosa River – also had greater ovarian transcript abundances for steroidogenic enzymes cytochrome P450 aromatase, 3β-hydroxysteroid dehydrogenase, and 17β-hydroxysteroid dehydrogenase at 24 °C compared to 34 °C, as well as higher liver mRNA levels of vitellogenins and choriogenins at cooler temperature. Transcript abundances for estrogen receptors esr1, esr2a, and esr2b were reduced at 34 °C in Amargosa River females, but not in Tecopa Bore females. T3 augmented gonadal gene transcript levels for steroid acute regulatory protein (StAR) transporter in both sexes and populations. T3 also downregulated liver estrogen receptor mRNAs in females from the warmer Tecopa Bore habitat only, suggesting T3 modulation of liver E2 sensitivity as a possible mechanism whereby temperature-induced changes in TH status may contribute to shifts in thermal sensitivity for oogenesis.

Per1/Per2 Disruption Reduces Testosterone Synthesis and Impairs Fertility in Elderly Male Mice.

Circadian rhythm disorders caused by genetic or environmental factors lead to decreased male fertility but the mechanisms are poorly understood. The current study reports that the mechanism of Per1/Per2 Double knockout (DKO) reduced the reproductive capacity of elderly male mice. The sperm motility and spermatogenic capacity of male DKO mice were weak. Hormone-targeted metabolomics showed reduced plasma levels of free testosterone in DKO male mice compared with WT male mice. Transcriptomic analysis of testicular tissue showed the down-regulation of testosterone synthesis-related enzymes (Cyp11a1, Cyp17a1, Hsd17b3, Hsd3b1, and Star) in the steroid hormone synthesis pathway. Spermatogenesis genes, Tubd1 and Pafah1b were down-regulated, influencing tubulin dynamics and leading to impaired motility. Seleno-compound metabolic loci, Scly and Sephs2, were up-regulated and Slc7a11 and Selenop were down-regulated. Western-blotting showed that steroid acute regulatory protein (StAR) and p-CREB, PKA and AC1 were reduced in testicular tissue of DKO mice compared to WT. Therefore, Per1/Per2 disruption reduced testosterone synthesis and sperm motility by affecting the PKA-StAR pathway, leading to decreased fertility.

Expression of kisspeptin and its receptor in different functional classes of ovarian follicle in the buffalo (Bubalus bubalis)

Recently, we reported the differential expression of kisspeptinergic system in the bubaline hypothalamus and corpus luteum. Here, we document the expression of kisspeptin (Kp) and its receptor (Kiss1r) in the ovarian follicles of the buffalo with respect to the functional status. Follicles of ≥10 to ≤13 mm diameter (n = 45) were retrospectively categorized into active (n = 18), intermediate (n = 16) and atretic (n = 11) follicles based on the concentrations of intrafollicular progesterone (P4) and estradiol (E2). The P4:E2 ratio was significantly lower in the active follicle (0.43 ± 0.08) than that of the intermediate (3.46 ± 0.53) and atretic (28.4 ± 10.6) follicles (P < 0.05). Relative fold change in the transcripts of kisspeptin (Kiss1), Kiss1r, gonadotrophin receptors, steroid acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), cytochrome P450 Family 19 subfamily A member 1 (CYP19A1), insulin like growth factor −1 (IGF-1), apoptotic factors (caspase 3 and B-cell lymphoma 2, BCL2) was calculated using qPCR in the follicular wall of the three categories of follicle (n = 8/group). In another experiment, histological sections of the ovary (n = 41) were used to group the follicles as described above and immunostaining of Kp, Kiss1r and aromatase was done. A significant upregulation of StAR, CYP11A1 and CYP19A1 in the active follicles supported the endocrine basis of follicular classification. The transcripts of Kiss1 and Kiss1r were upregulated by 19.45 fold and 4.25 fold, respectively in the active follicle as compared to other groups. Immunolocalization studies revealed that Kp and Kiss1r were localized to the basal and antral granulosa cells (GC) of the active and intermediate follicles; however, the staining intensity was stronger in the former group. Strong expression of CYP19A1 in the GC layer of active follicle supported the histological basis of defining the functional status of the follicle. It is concluded that the follicular compartment of the bubaline ovary expressed the constituents of kisspeptinergic system. The expression of Kp and Kiss1r was influenced by the functional status of the follicle with intense localization in the GC layer of the active follicles.

Widespread alterations to hypothalamic-pituitary-gonadal (HPG) axis signaling underlie high temperature reproductive inhibition in the eurythermal sheepshead minnow (Cyprinodon variegatus)

Fish experiencing abnormally high or prolonged elevations in temperature can exhibit impaired reproduction, even for species adapted to warm water environments. Such high temperature inhibition of reproduction has been linked to diminished gonadal steroidogenesis, but the mechanisms whereby hypothalamic-pituitary-gonadal (HPG) axis signaling is impacted by high temperature are not fully understood. Here, we characterized differences in HPG status in adult sheepshead minnow (Cyprinodon variegatus), a eurythermal salt marsh and estuarine species of eastern North America, exposed for 14 d to temperatures of 27 °C or 37 °C. Males and females at 37 °C had lower gonadosomatic index (GSI) values compared to fish at 27 °C, and females at 37 °C had fewer spawning capable eggs and lower circulating 17β-estradiol (E2). Gene transcripts encoding gonadotropin-inhibitory hormone (gnih) and gonadotropin-releasing hormone-3 (gnrh3) were higher in relative abundance in the hypothalamus of both sexes at 37 °C. While pituitary mRNAs for the β-subunits of follicle-stimulating hormone (fshβ) and luteinizing hormone (lhβ) were lowered only in males at 37 °C, Fsh and Lh receptor mRNA levels in the gonads were at lower relative levels in both the ovary and testis of fish at 37 °C. Females at 37 °C also showed reduced ovarian mRNA levels for steroid acute regulatory protein (star), P450 side-chain cleavage enzyme (cyp11a1), 3β-hydroxysteroid dehydrogenase (3βhsd), 17β-hydroxysteroid dehydrogenase (hsd17β3), and ovarian aromatase (cyp19a1a). Females at the higher 37 °C temperature also had a lower liver expression of mRNAs encoding estrogen receptor α (esr1) and several vitellogenin and choriogenin genes, but elevated mRNA levels for hepatic sex hormone-binding globulin (shbg). Our results substantiate prior findings that exposure of fish to high temperature can inhibit gonadal steroidogenesis and oogenesis, and point to declines in reproductive performance emerging from alterations at several levels of HPG axis signaling including increased hypothalamic Gnih expression, depressed gonadal steroidogenesis, and reduced egg yolk and egg envelope protein production in the liver.

Dietary supplementation of grape seed tannin extract stimulated testis development, changed fatty acid profiles and increased testis antioxidant capacity in pre-puberty hu lambs

Grape seed tannin extract (GPE) from wine grape pomace has many effective anti-oxidative effects and is used as a promising natural feed additive in the animal feed industry. This study investigated the effect of GPE as a source of tannin on the antioxidant capacity and testis development in Hu lambs. Twenty-seven 3-month-old ram lambs were randomly assigned to three groups. For each treatment group, nine lambs were allocated to nine pens (one lamb per pen). The lambs in the control group were fed a control diet without GPE for 61 days from D21 to D80. Group I (TAN1) was fed with 0.36% GPE diet, and Group II (TAN2) was fed with 0.72% GPE diet. After an 81-day feeding trial, all lambs except the heaviest and lightest in each group were humanely slaughtered and investigated. Results showed that feeding GPE did not affect the body weight, average daily gain, dry matter intake, scrotal circumference, and testis index. Meanwhile, feeding with 0.36% GPE diet increased testis weight, testis volume, and epididymis weight (P ≤ 0.05) compared with those of the control, but no difference was found between TAN1 and TAN2 groups. Copper-zinc superoxide dismutase (Cu-ZnSOD), steroid acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), follicle-stimulating hormone receptor (FSHR), elongation of very long chain fatty acid protein 2 (ELOVL2), fatty acid desaturase (FADS2), and proliferating cell nuclear antigen (PCNA) mRNA in TAN1 and TAN2 groups were significantly up-regulated (P < 0.05). GPE also markedly increased the antioxidant status of testis. Compared with the control group, the treatment groups showed significantly increased superoxide dismutase (SOD) activity (314.23 ± 18.64 U/mg prot in control, 505.22 ± 63.47 U/mg prot in TAN1 and 587.88 ± 55.94 U/mg prot in TAN2, P < 0.05) and total antioxidant capacity (T-AOC) (98.23 ± 18.99 U/g prot in control, 202.15 ± 34.19 U/g prot in TAN1 and 189.57 ± 18.95 U/g prot in TAN2, P < 0.05). Consuming 0.72% GPE also changed the fatty acid profiles in testis with increased C15:1, C22:6n3, and total n-3 fatty acids (P < 0.05) but decreased C22:5n3 (P < 0.05). Therefore, feeding lambs with GPE stimulated testis seminiferous tubule development and increased the number of Sertoli cells (10.56 ± 0.44 in control, 14.10 ± 0.57 in TAN1 and 13.60 ± 0.42, P < 0.05), and seminiferous tubule diameter (109.30 ± 4.56 μm in control, 164.49 ± 5.37 μm in TAN1 and 146.56 ± 4.53 μm in TAN2, P < 0.05). These results suggested that feeding GPE in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and polyunsaturated fatty acid metabolism-related genes, changed the fatty acid profiles, increased the antioxidant capacity in lamb's testis, and contributed to testis development and spermatogenesis.

Effect of feeding linseed diet on testis development, antioxidant capacity, and epididymal cauda sperm concentration in Chinese Hu lamb

Polyunsaturated fatty acids (PUFAs) are essential for mammalian testis development and sperm function. However, PUFAs that are contained in linseed oil are easily oxidized in the diet and biohydrogenated in the rumen. In this study, we investigated the effect of linseed as a source of PUFAs on the antioxidant capacity and testis development in Hu lamb. Seventy-five 3-month-old lambs were randomly assigned to three groups. Within each treatment group, 25 lambs were allocated to five pens (five lambs per pen). The lambs in the control group were fed a control diet without linseed for 42 days from D22 to D63. Group I (BS28) was fed a control diet from D22 to D35 and 8% linseed diet from D36 to D63. Group II (BS42) was fed an 8% linseed diet for 42 days from D22 to D63. After 63-day feeding trial, all lambs except the heaviest and lightest in each pen were humanely slaughtered and investigated. Results revealed that feeding linseed did not affect the body weight, scrotal circumference, and testis weight, whereas feeding linseed for 42 days increased the epididymis weight (37.85 ± 1.61 g vs. 32.09 ± 1.06 g, P < 0.05) compared with the control group. Feeding lambs with linseed for 42 days also significantly upregulated the expression of antioxidative (glutathione peroxidase 4 and copper-zinc superoxide dismutase), steroidogenesis (3β-hydroxysteroid dehydrogenase and steroid acute regulatory protein), and PUFA metabolism-related genes (fatty acid desaturase 2 and elongation of very long-chain fatty acid protein 2) and proliferating cell nuclear antigen mRNA (P < 0.05). It also increased the relative expression of mitochondrial DNA (P < 0.05), total antioxidant capacity (0.230 ± 0.019 mmol/mgprot vs. 0.175 ± 0.011 mmol/mgprot, P < 0.05), and superoxide dismutase (1661.467 ± 147.117 U/mgprot vs. 1158.891 ± 98.850 U/mgprot, P < 0.05) in testicular tissue but decreased the cholesterol concentration (0.331 ± 0.073 mmol/mgprot vs. 0.671 ± 0.092 mmol/mgprot, P < 0.05) compared with the control group. Therefore, feeding lambs with linseed for 42 days stimulated seminiferous tubule development and increased the number of Sertoli cells (20.71 ± 0.89 vs. 17.6 ± 0.73, P < 0.05), epididymal cauda lumina diameter (638.26 ± 22.32 μm vs. 444.41 ± 34.80 μm, P < 0.05), and the number of sperm in the epididymal cauda (68.91 ± 7.06 × 108/g vs. 36.61 ± 7.50 × 108/g). All these results suggested that feeding linseed in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and PUFA metabolism-related genes; increased the antioxidant capacity in lamb’s testis; and contributed to testis development and spermatogenesis.

Effect of dietary n-3 polyunsaturated fatty acid supplementation on the expression of genes involved in progesterone biosynthesis in the corpus luteum of goat (Capra hircus).

Emerging evidence indicates that dietary n-3 polyunsaturated fatty acids (PUFA) alter the fatty acid composition of corpus luteum (CL) and directly affect the luteal function in the cow, which is independent of the inhibitory effect on the endometrial PGF2α production. The present study, thus, investigated the effects of n-3 PUFA rich fish oil (FO) supplementation on the transcriptional modulation of genes involved in the biosynthesis of progesterone (P4 ) in the CL collected during the luteolytic phase of oestrous cycle in the goat. On the day of synchronized oestrus, goats (n=6/group) were fed an isocaloric diet supplemented with either FO or palm oil (PO). The dose of oil supplementation was 0.6mlkg-1 body weight, and the duration was 55-57days. The FO provided 156mgkg-1 body weight of n-3 eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The CL was collected by laparotomy on day 16 post-oestrus, and the relative abundance of P450 side-chain cleaving enzyme, steroid acute regulatory protein (StAR) and 3β-hydroxy steroid dehydrogenase (3β-HSD) genes was quantitated by real-time PCR. The results indicated that the dietary FO significantly upregulated the expression of 3β-HSD by 1.13-fold and downregulated StAR by ~2-fold as compared to PO group (p<.05). It is concluded that dietary FO differently affected the expression of genes involved in P4 synthesis in the CL during the luteolytic window of the oestrous cycle in the goat.

Lipoid congenital adrenal hyperplasia due to steroid acute regulatory protein (STAR) variants in Three Chinese patients

Congenital adrenal hyperplasia (CAH) encompasses a group of autosomal recessive diseases characterized by enzymatic defects in steroid synthesis. Lipoid congenital adrenal hyperplasia (LCAH) is the most severe form of CAH, insofar as the initial step of steroid synthesis is impaired. Variants in the steroid acute regulatory (STAR) gene are responsible for LCAH. To describe the clinical and genetic characteristics of three Chinese patients with LCAH. We analyzed the history, clinical manifestations, physical examination, laboratory data, and computed tomography findings of three girls with LCAH. The STAR gene of the probands and their parents were sequenced using genomic DNA. The wild-type and mutant STAR cDNAs were inserted into the pcDNA3.1(+) plasmid vector and transiently transfected into COS7 cells. The enzymatic activities of the wild-type and mutant STAR were evaluated by the enzyme-dependent conversion efficiency of cholesterol to pregnenolone. We identified the molecular genetic abnormalities in three patients with LCAH. All three patients had a female phenotype: karyotype of patients 1 and 2 was 46, XY and patient 3 was 46, XX. DNA sequencing revealed compound heterozygous variants in STAR for three probands. Two variants, c.659A > G/p.His220Arg and exon 2–3 deletion, were novel. In vitro functional studies uncovered that the His220Arg variant retained 19.2 % of enzymatic activity compared to that of the wild type.

PSXIII-33 Polyunsaturated fatty acid and their fetal programming effect on the FFARs from ewe lambs’ ovaries

Abstract Free fatty acids (FA) receptors (FFAR) are critical for metabolic functions and energy homeostasis. The FFAR1 and FFAR4 are activated by long-chain polyunsaturated FA, but FFAR2 and FFAR3 join to short-chain FA. There is no evidence fetal programming on nutritional effects on those receptors, insulin receptor (INS-R) and stereological enzyme in lambs’ ovaries. The objective of this study was to evaluate the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation to ewes during late gestation on finishing lamb ovary gene expression. Lambs born from ewes supplemented with Ca salts of EPA and DHA (PUFA), or palm FA distillate (PFAD) high in palmitic and oleic acid at 0.39% DM during the last 50d of gestation were used. After weaning and high concentrate diet adaptation (45d), 14 Hampshire x Dorset cross females lambs were blocked by initial BW and used in a 2x2 factorial arrangement of treatments using the factors of dam supplementation and lamb diets (PUFA or PFAD at 1.5% DM). At day 42, lambs were euthanized and ovary samples were obtained for FFARs, INS-R and steroid acute regulatory protein (STAR) mRNA concentration analysis. There was a dam diet effect on the abundance of FFAR4 mRNA (P = 0.05); lambs born from PUFA dams showed greater concentration than PFAD. Also, PUFA lambs tend to have greater concentration of FFAR4 (P = 0.07). The abundance of FFAR2, INS-R, and STAR mRNA was not significant for dam or lamb diet effect (P &amp;gt; 0.1). The mRNA of FFAR1 and FFAR3 were no found in the samples. However, mRNA concentration of FFAR2 and STAR were positively associated (r = 0.74 P &amp;lt; 0.05). In conclusion, dam and lamb diets modified FFAR4 mRNA concentration; and there might be a functional association between FFAR2 and STAR.

Effect of short-term nutritional supplementation on hormone concentrations in ovarian follicular fluid and steroid regulating gene mRNA abundances in granulosa cells of ewes

This study was conducted to investigate effects of short-term nutritional supplementation on concentrations of reproductive hormones in follicular fluid and on mRNA abundance in granulosa cells (GCs) during the luteal phase of ewes. Eighteen ewes were randomly assigned to treatment or control groups (n = 9, each group). All the ewes were subjected to estrous synchronization using a Controlled Intravaginal Releasing Device (CIDR). From the second to the eleventh day of estrous synchronization, ewes were fed a diet with a relatively greater (treatment group) or a maintenance (control group) energy content. Samples of follicular fluid and GCs were collected from antral follicles of estrous ewes after CIDR removal. The results indicate mean FSH concentration of fluid in small follicles and mean LH concentrations of fluid in large follicles of the ewes in the treatment group were greater (P < 0.05) than that of ewes in the control group. Follicular fluid E2 concentrations were directly related (P < 0.05) to follicular diameter. The ewes of the treatment group had greater (P < 0.05), compared with the control group, abundances of Follicle Stimulating Hormone Receptor (FSHR) in small and medium follicles, and (P<0.05) Luteinizing Hormone Receptor (LHR), Steroid Acute Regulatory Protein (STAR), cytochrome P450 (CYP17A1, CYP19A1) enzyme and Estrogen Receptor (ESR1) in large follicles. The results of this study provide evidence for a potential mechanism by which short-term nutritional supplementation improves follicular development possibly because of increased expression of steroid synthesis-regulating genes in large follicles.

Heat stress during the luteal phase decreases luteal size but does not affect circulating progesterone in gilts1.

Heat stress (HS) occurs when heat dissipation mechanisms are insufficient to maintain euthermia, and it is associated with seasonal infertility (SI), which manifests as smaller litters, longer wean-to-estrus interval, increased abortions, and reduced conception rates. To understand HS-induced mechanisms underlying SI, crossbred post-pubertal gilts (167 ± 10 kg; n = 14) experienced either thermal neutral (TN, 20 ± 1 °C, n = 7) or cyclical HS (35 ± 1 °C for 12 h and 31.6 °C for 12 h, n = 7) conditions from 2 to 12 d post-estrus (dpe). Estrous cycles were synchronized via altrenogest administration for 14 d, phenotypic manifestation of estrus was observed and gilts were assigned to experimental treatment. Gilts were limit fed 2.7 kg daily with ad libitum water access. Blood was collected at 0, 4, 8, and 12 dpe via jugular venipuncture and animals were humanely euthanized at 12 dpe. The corpora lutea (CL) width were measured via digital calipers on both ovaries, and CL from one ovary were excised, weighed, and protein and steroid abundance analyzed via western blotting and ELISA, respectively. Relative to TN, HS increased (P < 0.01) rectal temperature and respiration rates and reduced (P < 0.01) feed intake. The CL from HS ovaries were reduced in diameter (P < 0.05) and weight (P < 0.01) relative to those from TN animals. No difference (P = 0.38) in CL or serum progesterone concentrations between groups was observed at any time point, though at 12 dpe the serum progesterone:CL weight was increased (P < 0.10) by HS. No treatment differences (P = 0.84) in circulating insulin were observed. Luteal protein abundance of steroid acute regulatory protein, 3 beta-hydroxysteroid, or prostaglandin F2α receptor were not different between treatments (P = 0.73). Taken together, these data demonstrate that the CL mass is HS sensitive, but this phenotype does not appear to be explained by the metrics evaluated herein. Regardless, HS-induced decreased CL size may have important implications to pig SI and warrants additional attention.

Endocrine and reproductive responses of male and female cattle to agonists of gonadotrophin-releasing hormone

The pituitary response in cattle to treatment with GnRH agonist has two phases. In the acute phase secretion of LH is increased, while the chronic phase is characterized by a downregulation of GnRH receptors and insensitivity of gonadotrophs to natural sequence GnRH. After long-term treatment with GnRH agonist, cattle do not have pulsatile secretion of LH but maintain basal LH. This is associated with reduced pituitary contents of LH, LH mRNA, FSH and FSH mRNA. Long-term treatment of bulls with GnRH agonist results in an increase in testicular LH receptors and high plasma testosterone. Heifers treated with a GnRH agonist from early in the oestrous cycle develop a larger corpus luteum and secrete more progesterone. Increased steroidogenesis is reflected in increased steroid acute regulatory (StAR) protein and steroidogenic enzymes in the testes and corpus luteum. GnRH agonists have potential as novel strategies for reproductive management in cattle. A GnRH agonist bioimplant was recently used to block the LH surge after FSH stimulation of follicle growth in heifers. Ovulation was induced by injection of LH, and heifers were inseminated relative to the LH injection. This GnRH agonist-LH protocol provides a model for studying the gonadotrophin requirements for follicular growth and oocyte maturation in cattle, and will enable controlled in vivo maturation of oocytes before recovery for in vitro procedures.

Human scalp hair: Modulation by various factors and hormones do estrogens inhibit or stimulate-A perplexing perspective.

Several journal reports, reviews, and commentaries over the last 20-25years have pointed out the controversy attached to 17β-estradiol's inhibitory or stimulatory influence on hair follicle growth/cycling citing rodent (murine) and human results. While 17β-estradiol is the most potent sex steroid hormone in the body and has almost equal affinity for estrogen receptor (ER) alpha (α) and beta (β), there appears to be specific ER-mediated effects on scalp hair follicles/growth, etc. Additionally, the newly discovered G protein-coupled estrogen receptor (GPR30 or GPER) and the orphan receptor, estrogen-related receptor (ERR) gamma (γ), in skin and other tissue sites have potential impacts of how estrogens via these receptors may alter scalp hair characteristics, but this remains to be elucidated. Conversely, the negative impact of the 5α-reductase enzyme and its steroid product, 5α-dihydrotestosterone, on scalp hair growth is clear. Less clear is how 17β-estradiol is stimulatory in some scalp hair studies, but inhibitory in others. This brief summary examines the potential influences of steroidogenesis via aromatase (estrogen biosynthesis) and 5α-reductase expression, their enzyme activities, and steroid products along with the concepts of how steroid acute regulatory protein (StAR) and estrone sulfate may be involved in the complex hormonal, cellular/molecular signaling cascade of the hair follicle in growth and cycling.

Impact of heat stress during the follicular phase on porcine ovarian steroidogenic and phosphatidylinositol-3 signaling.

Environmental conditions that impede heat dissipation and increase body temperature cause heat stress (HS). The study objective was to evaluate impacts of HS on the follicular phase of the estrous cycle. Postpubertal gilts (126.0 ± 21.6 kg) were orally administered altrenogest to synchronize estrus, and subjected to either 5 d of thermal-neutral (TN; 20.3 ± 0.5 °C; n = 6) or cyclical HS (25.4 - 31.9 °C; n = 6) conditions during the follicular phase preceding behavioral estrus. On d 5, blood samples were obtained, gilts were euthanized, and ovaries collected. Fluid from dominant follicles was aspirated and ovarian protein homogenates prepared for protein abundance analysis. HS decreased feed intake (22%; P = 0.03) and while plasma insulin levels did not differ, the insulin:feed intake ratio was increased 3-fold by HS (P = 0.02). Insulin receptor protein abundance was increased (29%; P < 0.01), but insulin receptor substrate 1, total and phosphorylated protein kinase B, superoxide dismutase 1, and acyloxyacyl hydrolase protein abundance were unaffected by HS (P > 0.05). Plasma and follicular fluid 17β-estradiol, progesterone, and lipopolysaccharide-binding protein concentrations as well as abundance of steroid acute regulatory protein, cytochrome P450 19A1, and multidrug resistance-associated protein 1 were not affected by HS (P > 0.05). HS increased estrogen sulfotransferase protein abundance (44%; P = 0.02), toll-like receptor 4 (36%; P = 0.05), and phosphorylated REL-associated protein (31%; P = 0.02). Regardless of treatment, toll-like receptor 4 protein was localized to mural granulosa cells in the porcine ovary. In conclusion, HS altered ovarian signaling in postpubertal gilts during their follicular phase in ways that likely contributes to seasonal infertility.

Disturbances in reproduction and expression of steroidogenic enzymes in aquatic invertebrates exposed to components of the herbicide Roundup

Exposure of organisms to environmental contaminants is a growing concern. We have investigated the effects of the individual active ingredients of the herbicide Roundup (glyphosate and diquat dibromide [DD]) since Roundup causes alterations in reproduction, mortality, and development in the aquatic snail Lymnaea palustris. Snails chronically treated with elevated but ecologically relevant levels of DD exhibit reduction in fecundity ( p &lt; 0.05), while fecundity in glyphosate-treated snails is comparable to or exceeds control levels. To investigate a possible mechanism for the reproductive disturbance, we monitored levels of steroid acute regulatory (StAR) protein in whole snails and observed a correlation in StAR protein decrease with treatment with Roundup, glyphosate, or DD. We detect StAR in organs where steroid biosynthesis occurs (ovotestis, brain, kidney); StAR protein is reduced following chronic exposure to Roundup, glyphosate, or DD ( p &lt; 0.01). Estradiol and testosterone concentrations in hemolymph were measured by enzyme-linked immunosorbent assay following 3-week exposure of snails to 3.5 mg/L glyphosate or 140 µg/L DD. Testosterone levels decrease in DD-treated groups ( p &lt; 0.05); a trend of lower testosterone is also observed in glyphosate-treated groups ( p &gt; 0.05). Estradiol concentration is greater than or equal to control levels in glyphosate, but decreased in DD ( p &lt; 0.05). Because of its role in the conversion of testosterone to estradiol, we monitored abundance of aromatase and observed a reduction ( p &lt; 0.05) in DD-treated snails (consistent with the drop in fecundity and estradiol levels) and a comparable level to control in glyphosate-treated snails (consistent with their high fecundity and estradiol levels). Although the toxicity of commercially-available Roundup to aquatic animals may have many contributing factors including its inactive surfactant, the constituent of Roundup associated with the greatest reproductive disturbances and observed developmental abnormalities of offspring is DD. This study details the analysis of particular herbicide constituents and their effect on specific targets in the reproductive pathway.

Kisspeptin regulates ovarian steroidogenesis during delayed embryonic development in the fruit bat, Cynopterus sphinx.

Cynopterus sphinx, a fruit bat, undergoes delayed embryonic development during the winter months, a period that corresponds to low levels of progesterone and estradiol synthesis by the ovary. Kisspeptins (KPs) are a group of neuropeptide hormones that act via G-protein coupled receptor 54 (GPR54) to stimulate hypothalamic secretion of Gonadotropin-releasing hormone, thereby regulating ovarian steroidogenesis, folliculogenesis, and ovulation. GPR54 is also expressed in the ovary, suggesting a direct role for KPs in ovarian steroidogenesis. The aim of present study was to determine if a low serum level of KP is responsible for reduced progesterone and estradiol levels during the period of delayed embryonic development in C. sphinx. Indeed, low serum KP abundance corresponded to reduced expression of GPR54 in ovarian luteal cells during the period of delayed development compared to normal development. In vitro and in vivo treatment with KP increased GPR54 abundance, via Extracellular signal regulated kinase and its downstream mediators, leading to increased progesterone synthesis in the ovary during delayed embryonic development. KP treatment also increased cholesterol uptake and elevated expression of Luteinizing hormone receptor and Steroid acute regulatory protein in the ovary, suggesting that elevation in circulating KP during delayed embryonic development may reactivate luteal activity. KPs may also enhance cell survival (BCL-2, reduced Caspase 3 activity) and angiogenesis (Vascular endothelium growth factor) during this period. The findings of this study thus demonstrate a regulatory role for KPs in the maintenance of luteal steroidogenesis during pregnancy in C. sphinx.

Presence of Germ Cells in Disorders of Sex Development: Implications for Fertility Potential and Preservation

We sought to determine the presence of germ cells in the gonads of patients with disorders of sex development to establish whether preservation of germ cells for future fertility potential is possible. We hypothesized that germ cells are present but vary by age and diagnosis. We reviewed histology from patients with disorders of sex development who underwent gonadectomy/biopsy from 2002 to 2014 at a single institution for pathological classification of the gonad, composition of gonadal stroma and germ cell presence. A total of 44 patients were identified and germ cells were present in 68%. The presence and average number of germ cells per mm2 were analyzed by gonad type and diagnosis. By gonad type all ovotestes, most testes, ovaries and dysgenetic testes, and 15% of streak gonads had germ cells present. By diagnosis germ cells were present in all patients with complete androgen insensitivity syndrome, Denys-Drash syndrome, SRY mutation, mixed gonadal dysgenesis, ovotesticular conditions and StAR (steroid acute regulatory protein) deficiency, in some patients with persistent müllerian duct syndrome, XO/XY Turner syndrome and disorders of sex development not otherwise specified, and in none with complete or partial gonadal dysgenesis. Germ cells were present in the gonads of 88% of patients 0 to 3 years old, 50% of those 4 to 11 years old and 43%of those older than 12 years. Germ cells were present in the majority of our cohort and the presence decreased with age. This novel, fertility driven evaluation of germ cell quantity in a variety of disorders of sex development suggests that fertility potential may be greater than previously thought. Further studies must be done to evaluate a larger population and examine germ cell quality to determine the viability of these germ cells.