Characterizing the neuroendocrine and ovarian defects of androgen receptor-knockout female mice

2013 ◽  
Vol 305 (6) ◽  
pp. E717-E726 ◽  
Author(s):  
Xiaobing B. Cheng ◽  
Mark Jimenez ◽  
Reena Desai ◽  
Linda J. Middleton ◽  
Shai R. Joseph ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Negative Feedback ◽  
Positive Feedback ◽  
Growth Patterns ◽  
Corpora Lutea ◽  
Preovulatory Follicle ◽  
Lh Surge ◽  
Antral Follicles ◽  
Female Mice ◽  
Follicle Diameter

Homozygous androgen receptor (AR)-knockout (ARKO) female mice are subfertile due to both intra- and extraovarian (neuroendocrine) defects as defined by ovary transplantation. Using ARKO mice, this study set out to reveal the precise AR-regulated pathways required for optimal androgen-regulated ovulation and fertility. ARKO females exhibit deficient neuroendocrine negative feedback, with a reduced serum luteinizing hormone (LH) response to ovariectomy (OVX) ( P < 0.01). Positive feedback is also altered as intact ARKO females, at late proestrus, exhibit an often mistimed endogenous ovulatory LH surge. Furthermore, at late proestrus, intact ARKO females display diminished preovulatory serum estradiol (E2; P < 0.01) and LH ( P < 0.05) surge levels and reduced Kiss1 mRNA expression in the anteroventral periventricular nucleus ( P < 0.01) compared with controls. However, this reduced ovulatory LH response in intact ARKO females can be rescued by OVX and E2 priming or treatment with endogenous GnRH. These findings reveal that AR regulates the negative feedback response to E2, E2-positive feedback is compromised in ARKO mice, and AR-regulated negative and positive steroidal feedback pathways impact on intrahypothalamic control of the kisspeptin/GnRH/LH cascade. In addition, intraovarian AR-regulated pathways controlling antral to preovulatory follicle dynamics are disrupted because adult ARKO ovaries collected at proestrus have small antral follicles with reduced oocyte/follicle diameter ratios ( P < 0.01) and increased proportions of unhealthy large antral follicles ( P < 0.05) compared with controls. As a consequence of aberrant follicular growth patterns, proestrus ARKO ovaries also exhibit fewer preovulatory follicle ( P < 0.05) and corpora lutea numbers ( P < 0.01). However, embryo development to the blastocyst stage is unchanged in ARKO females, and hence, the subfertility is a consequence of reduced ovulations and not altered embryo quality. These findings reveal that the AR has a functional role in neuroendocrine regulation and timing of the ovulatory LH surge as well as antral/preovulatory follicle development.

124 RELATIONSHIPS BETWEEN PREOVULATORY FOLLICLE AND CORPUS LUTEUM BLOOD FLOW IN MARES

2012 ◽  
Vol 24 (1) ◽  
pp. 174
Author(s):  
A. Wischral ◽  
K. T. Haag ◽  
G. R. Fonseca ◽  
M. O. Gastal ◽  
S. S. King ◽  
...  
Keyword(s):  
Blood Flow ◽  
Doppler Ultrasonography ◽  
New Technologies ◽  
Power Doppler ◽  
Corpora Lutea ◽  
Power Doppler Ultrasonography ◽  
Preovulatory Follicle ◽  
Vascular Perfusion ◽  
Follicle Diameter ◽  
Student’S T

Colour- and power-Doppler ultrasonography have recently been used as potential new technologies to assess the degree of vascular perfusion of the ovary and follicles for research and clinical studies of ovarian and follicle hemodynamics and to predict fertility in horses, cattle and humans. In the present study, the following hypotheses were tested: (1) preovulatory follicle (POF) diameter (≥30 mm), but not blood flow, is repeatable between cycles within the same mare; (2) POF diameter and blood flow are good indicators of follicle status; (3) double POF have similar blood flow; and (4) highly vascularized POF produce corpora lutea (CL) with greater blood flow. Non-lactating mares (n = 13; 5 to 21 years old) of mixed breeds were used from March to May in the Northern Hemisphere. Follicle diameter and vascularity of the follicle wall before the first and second ovulations of the season and vascularity of the first CL were measured daily using transrectal colour-Doppler ultrasonography. The vascularity of the follicle wall and CL was based on the display of the blood-flow signals visualised in a slow, continuous-motion evaluation. Statistical analyses were performed by the SAS MIXED procedure, ANOVA and Student's t-tests and Spearman's correlation. A total of 26 periovulatory periods were evaluated. Unexpectedly, there were 84.6% (11/13) and 61.5% (8/13) double dominant POF and 30.8% (4/13) and 46.2% (6/13) double ovulations in the first and second periovulatory periods, respectively. The POF diameters were highly correlated (r = 0.68; P < 0.0001) between the first and second periovulatory periods. The diameter of the POF 5 days before the first ovulation was larger (P < 0.004) than before the second ovulation of the year. However, the POF vascularity did not differ between those periods. For 4 days before ovulation (Day 0), the diameter and blood flow of the POF were greater (P < 0.05) than for those follicles that underwent atresia in single- and double-ovulatory mares. The POF diameter and blood flow were positively correlated in ovulatory (r = 0.51; P < 0.0001) and in atretic (r = 0.32; P < 0.02) follicles. In double-ovulatory mares, POF diameter and blood flow increased (P < 0.0006) for 5 days before ovulation, with no difference between the 2 follicles in the same cycle for each parameter. The POF blood flow was positively correlated (r = 0.32; P < 0.0009) with CL vascularity during the first periovulatory period (Day –7 to +6) of the season. Furthermore, a positive correlation (r = 0.58; P < 0.01) was observed between the maximum vascularity of the POF and its subsequent CL. In conclusion, although preliminary, our results demonstrated that (a) POF vascularity is not repeatable within individuals; (b) potential atretic POF have low blood flow; (c) double POF have similar vascularity; and (d) greater blood flow to the POF is associated with higher CL vascularity.

scholarly journals Absent Progesterone Signaling in Kisspeptin Neurons Disrupts the LH Surge and Impairs Fertility in Female Mice

Endocrinology ◽  
2015 ◽  
Vol 156 (9) ◽  
pp. 3091-3097 ◽  
Author(s):  
Shannon B. Z. Stephens ◽  
Kristen P. Tolson ◽  
Melvin L. Rouse ◽  
Matthew C. Poling ◽  
Minako K. Hashimoto-Partyka ◽  
...  
Keyword(s):  
Positive Feedback ◽  
Progesterone Receptors ◽  
Sex Steroid ◽  
Corpora Lutea ◽  
Neuronal Activation ◽  
Estrogen Regulation ◽  
Lh Surge ◽  
Normal Fertility ◽  
Periventricular Nucleus

Kisspeptin, encoded by Kiss1, stimulates GnRH neurons to govern reproduction. In rodents, estrogen-sensitive kisspeptin neurons in the anterior ventral periventricular nucleus and neighboring periventricular nucleus are thought to mediate sex steroid-induced positive feedback induction of the preovulatory LH surge. These kisspeptin neurons coexpress estrogen and progesterone receptors and display enhanced neuronal activation during the LH surge. However, although estrogen regulation of kisspeptin neurons has been well studied, the role of progesterone signaling in regulating kisspeptin neurons is unknown. Here we tested whether progesterone action specifically in kisspeptin cells is essential for proper LH surge and fertility. We used Cre-lox technology to generate transgenic mice lacking progesterone receptors exclusively in kisspeptin cells (termed KissPRKOs). Male KissPRKOs displayed normal fertility and gonadotropin levels. In stark contrast, female KissPRKOs displayed earlier puberty onset and significant impairments in fertility, evidenced by fewer births and substantially reduced litter size. KissPRKOs also had fewer ovarian corpora lutea, suggesting impaired ovulation. To ascertain whether this reflects a defect in the ability to generate sex steroid-induced LH surges, females were exposed to an estradiol-positive feedback paradigm. Unlike control females, which displayed robust LH surges, KissPRKO females did not generate notable LH surges and expressed significantly blunted cfos induction in anterior ventral periventricular nucleus kisspeptin neurons, indicating that progesterone receptor signaling in kisspeptin neurons is required for normal kisspeptin neuronal activation and LH surges during positive feedback. Our novel findings demonstrate that progesterone signaling specifically in kisspeptin cells is essential for the positive feedback induction of normal LH surges, ovulation, and normal fertility in females.

scholarly journals Female Mice Haploinsufficient for an Inactivated Androgen Receptor (AR) Exhibit Age-Dependent Defects That Resemble the AR Null Phenotype of Dysfunctional Late Follicle Development, Ovulation, and Fertility

Endocrinology ◽  
2007 ◽  
Vol 148 (8) ◽  
pp. 3674-3684 ◽  
Author(s):  
K. A. Walters ◽  
C. M. Allan ◽  
M. Jimenez ◽  
P. R. Lim ◽  
R. A. Davey ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Ovarian Function ◽  
Gene Dosage ◽  
Early Embryo ◽  
Ovulation Rate ◽  
Corpora Lutea ◽  
Cell Stage ◽  
Gene Dosage Effect ◽  
Female Mice ◽  
Age Dependent

The role of classical genomic androgen receptor (AR) mediated actions in female reproductive physiology remains unclear. Female mice homozygous for an in-frame deletion of exon 3 of the Ar (AR−/−) were subfertile, exhibiting delayed production of their first litter (AR+/+ = 22 d vs. AR−/− = 61 d, P &lt; 0.05) and producing 60% fewer pups/litter (AR+/+: 8.1 ± 0.4 vs. AR−/−: 3.2 ± 0.9, P &lt; 0.01). Heterozygous females (AR+/−) exhibited an age-dependent 55% reduction (P &lt; 0.01) in pups per litter, evident from 6 months of age (P &lt; 0.05), compared with AR+/+, indicating a significant gene dosage effect on female fertility. Ovulation was defective with a significant reduction in corpora lutea numbers (48–79%, P &lt; 0.01) in 10- to 12- and 26-wk-old AR+/− and AR−/− females and a 57% reduction in oocytes recovered from naturally mated AR−/− females (AR+/+: 9.8 ± 1.0 vs. AR−/−: 4.2 ± 1.2, P &lt; 0.01); however, early embryo development to the two-cell stage was unaltered. The delay in first litter, reduction in natural ovulation rate, and aromatase expression in AR+/− and AR−/− ovaries, coupled with the restored ovulation rate by gonadotropin hyperstimulation in AR−/− females, suggest aberrant gonadotropin regulation. A 2.7-fold increase (AR+/+: 35.4 ± 13.4 vs. AR−/−: 93.9 ± 6.1, P &lt; 0.01) in morphologically unhealthy antral follicles demonstrated deficiencies in late follicular development, although growing follicle populations and growth rates were unaltered. This novel model reveals that classical genomic AR action is critical for normal ovarian function, although not for follicle depletion and that haploinsufficiency for an inactivated AR may contribute to a premature reduction in female fecundity.

scholarly journals 11β-Hydroxysteroid dehydrogenase expression and activities in bovine granulosa cells and corpora lutea implicate corticosteroids in bovine ovarian physiology

2007 ◽  
Vol 193 (2) ◽  
pp. 299-310 ◽  
Author(s):  
L M Thurston ◽  
D R E Abayasekara ◽  
A E Michael
Keyword(s):  
Molecular Mass ◽  
Granulosa Cells ◽  
Luteal Phase ◽  
Hydroxysteroid Dehydrogenase ◽  
Follicular Phase ◽  
Corpora Lutea ◽  
Luteal Cells ◽  
Antral Follicles ◽  
Follicle Diameter ◽  
Dependent Type

Cortisol–cortisone metabolism is catalysed by the bi-directional NADP(H)-dependent type 1 11β-hydroxysteroid dehydrogenase (11βHSD1) enzyme and the oxidative NAD+-dependent type 2 11βHSD (11βHSD2). This study related the expression of 11βHSD1 and 11βHSD2 enzymes (mRNA and protein) to net 11-ketosteroid reductase and 11β-dehydrogenase (11β-DH) activities in bovine follicular granulosa and luteal cells. Granulosa cells were isolated from follicles of < 4, 4–8, > 8 and > 12 mm in diameter in either the follicular or luteal phase of the ovarian cycle. Luteal cells were obtained from corpora lutea (CL) in the early non-pregnant luteal phase. Enzyme expression was assessed by reverse transcription-PCR and western blotting, while enzyme activities were measured over 1 h in cell homogenates using radiometric conversion assays with 100 nM [3H]cortisone or [3H]cortisol and pyridine dinucleotide cofactors. Irrespective of follicle diameter, the expression of 11βHSD2 and NAD+-dependent oxidation of cortisol predominated in granulosa cells harvested in the follicular phase. In contrast, in granulosa cells obtained from luteal phase follicles and in bovine luteal cells, expression of 11βHSD1 exceeded that of 11βHSD2 and the major enzyme activity was NADP+-dependent cortisol oxidation. Increasing follicular diameter was associated with progressive increases in expression and activities of 11βHSD2 and 11βHSD1 in follicular and luteal phase granulosa cells respectively. In follicular phase granulosa cells from antral follicles < 12 mm, 11βHSD1 migrated with a molecular mass of 34 kDa, whereas in the dominant follicle, CL and all luteal phase granulosa cells, a second protein band of 68 kDa was consistently detected. In all samples, 11βHSD2 had a molecular mass of 48 kDa, but in large antral follicles (> 8 mm), there was an additional immunoreactive band at 50 kDa. We conclude that 11βHSD2 is the predominant functional 11βHSD enzyme expressed in follicular phase granulosa cells from growing bovine antral follicles. In contrast, in bovine granulosa cells from dominant or luteal phase follicles, and in bovine luteal cells from early non-pregnant CL, 11βHSD1 is the major glucocorticoid-metabolising enzyme. The increasing levels of cortisol inactivation by the combined NADP+- and NAD+-dependent 11β-DH activities suggest a need to restrict cortisol access to corticosteroid receptors in the final stages of follicle development.

scholarly journals Ovarian Expression of Insulin-Like Peptide 3 (INSL3) and Its Receptor (RXFP2) During Development of Bovine Antral Follicles and Corpora Lutea and Measurement of Circulating INSL3 Levels During Synchronized Estrous Cycles

Endocrinology ◽  
2013 ◽  
Vol 154 (5) ◽  
pp. 1897-1906 ◽  
Author(s):  
Leanne Satchell ◽  
Claire Glister ◽  
Emma C. Bleach ◽  
Richard G. Glencross ◽  
Andrew B. Bicknell ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Major Product ◽  
Corpora Lutea ◽  
Potential Contribution ◽  
Mrna Levels ◽  
Estrous Cycles ◽  
Lh Surge ◽  
Antral Follicles

Abstract Insulin-like peptide 3 (INSL3), a major product of testicular Leydig cells, is also expressed by the ovary, but its functional role remains poorly understood. Here, we quantified expression of INSL3 and its receptor RXFP2 in theca interna cell (TIC) and granulosa cell compartments of developing bovine antral follicles and in corpora lutea (CL). INSL3 and RXFP2 mRNA levels were much higher in TIC than granulosa cell and increased progressively during follicle maturation with INSL3 peaking in large (11-18 mm) estrogen-active follicles and RXFP2 peaking in 9- to 10-mm follicles before declining in larger (11-18 mm) follicles. Expression of both INSL3 and RXFP2 in CL was much lower than in TIC. In situ hybridization and immunohistochemistry confirmed abundant expression of INSL3 mRNA and protein in TIC. These observations indicate follicular TIC rather than CL as the primary site of both INSL3 production and action, implying a predominantly autocrine/paracrine role in TIC. To corroborate the above findings, we showed that in vitro exposure of TIC to a luteinizing concentration of LH greatly attenuated expression of both INSL3 and its receptor while increasing progesterone secretion and expression of STAR and CYP11A1. Moreover, in vivo, a significant cyclic variation in plasma INSL3 was observed during synchronized estrous cycles. INSL3 and estradiol-17β followed a similar pattern, both increasing after luteolysis, before falling sharply after the LH surge. Thus, theca-derived INSL3, likely from the dominant preovulatory follicle, is detectable in peripheral blood of cattle, and expression is down-regulated during luteinization induced by the preovulatory LH surge. Collectively, these findings underscore the likely role of INSL3 as an important intrafollicular modulator of TIC function/steroidogenesis, while raising doubts about its potential contribution to CL function.

scholarly journals Positive, But Not Negative Feedback Actions of Estradiol in Adult Female Mice Require Estrogen Receptor α in Kisspeptin Neurons

Endocrinology ◽  
2015 ◽  
Vol 156 (3) ◽  
pp. 1111-1120 ◽  
Author(s):  
Sharon L. Dubois ◽  
Maricedes Acosta-Martínez ◽  
Mary R. DeJoseph ◽  
Andrew Wolfe ◽  
Sally Radovick ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Adult Female ◽  
Negative Feedback ◽  
Positive Feedback ◽  
Estrogen Receptor Α ◽  
Female Mice ◽  
Lh Secretion ◽  
Express Estrogen Receptor ◽  
Specific Deletion

Abstract Hypothalamic kisspeptin (Kiss1) neurons express estrogen receptor α (ERα) and exert control over GnRH/LH secretion in female rodents. It has been proposed that estradiol (E2) activation of ERα in kisspeptin neurons in the arcuate nucleus (ARC) suppresses GnRH/LH secretion (negative feedback), whereas E2 activation of ERα in kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) mediates the release of preovulatory GnRH/LH surges (positive feedback). To test these hypotheses, we generated mice bearing kisspeptin cell–specific deletion of ERα (KERαKO) and treated them with E2 regimens that evoke either negative or positive feedback actions on GnRH/LH secretion. Using negative feedback regimens, as expected, E2 effectively suppressed LH levels in ovariectomized (OVX) wild-type (WT) mice to the levels seen in ovary-intact mice. Surprisingly, however, despite the fact that E2 regulation of Kiss1 mRNA expression was abrogated in both the ARC and AVPV of KERαKO mice, E2 also effectively decreased LH levels in OVX KERαKO mice to the levels seen in ovary-intact mice. Conversely, using a positive feedback regimen, E2 stimulated LH surges in WT mice, but had no effect in KERαKO mice. These experiments clearly demonstrate that ERα in kisspeptin neurons is required for the positive, but not negative feedback actions of E2 on GnRH/LH secretion in adult female mice. It remains to be determined whether the failure of KERαKO mice to exhibit GnRH/LH surges reflects the role of ERα in the development of kisspeptin neurons, in the active signaling processes leading to the release of GnRH/LH surges, or both.

scholarly journals Musashi as a Regulator of the Follicle-Stimulating Hormone in the Gonadotropes

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A545-A545
Author(s):  
Ana Rita Silva Moreira ◽  
Alexandra Lagasse ◽  
Anessa Haney ◽  
Ulrich Boehm ◽  
Michael G Kharas ◽  
...  
Keyword(s):  
Protein Content ◽  
Follicle Stimulating Hormone ◽  
Mrna Translation ◽  
Reproductive Hormones ◽  
Corpora Lutea ◽  
Control Female ◽  
Lh Surge ◽  
Female Mice ◽  
Qrt Pcr ◽  
Stimulating Hormone

Abstract The cyclic expression of gonadotropin releasing-hormone receptors (GnRHR), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) by pituitary gonadotropes is critical in the female reproductive process. We have shown that the translational regulator Musashi (MSI) binds to Gnrhr mRNA and inhibits its translation, and the gonadotrope-specific deletion of Msi1 and Msi2 (Gon-Msi-null) leads to increased pituitary GnRHR protein levels. An in silico analysis of gonadotropin mRNAs revealed 5 different MSI binding elements in the 3’UTR of Fshb mRNA. We hypothesize that, in addition to Gnrhr, MSI may also bind and repress Fshb mRNA translation in the gonadotropes. To test if MSI does target the Fshb transcript in the pituitary, we performed RNA immunoprecipitation (IP) on pooled control female mouse pituitaries using a MSI1 antibody and measured Fshb mRNA by qRT-PCR. To study the in vivo effects of MSI on Fshb, we harvested the pituitaries of the Gon-Msi-null (MUT) female mice and their littermate controls (CTL) during the estrous cycle. We collected serum and protein for EIAs to measure the levels of FSH and LH, and RNA for Fshb qRT-PCR. We harvested proestrous ovaries and fixed them for embedding, sectioning, and H&E staining. Our RNA IP experiments show a 7-fold enrichment for Fshb with the MSI1 antibody. The Gon-Msi-null females have significantly higher pituitary FSH protein content than controls on estrous morning (MUT: 4.8±1.3 vs. CTL: 1.8±2.6 ng/ml/μg protein, p&lt;0.0001, n=9-10/group). These mice also have increased serum FSH levels (MUT: 56.9±6.4 vs. CTL: 44.5±9.6 ng/ml, p=0.0147, n=9-10/group). No changes at the Fshb mRNA level were detected. Analysis of Gon-Msi-null ovaries revealed a 50% decrease in the number of follicles, with significant decreases in the average numbers of maturing follicles (p&lt;0.0175) and corpora lutea (p&lt;0.0215). Interestingly, the LH levels in these mice were also altered. The Gon-Msi-null females show a decrease in the pituitary LH protein content in the evening of proestrus (MUT: 11.8±1.4 vs. CTL: 15.1±2.0 ng/ml/μg protein, p=0.0333, n=7/group), in addition to a delayed and blunted LH surge (MUT: 2.6±1.9 vs. CTL: 7.3±3.5 ng/ml, p=0.0089, n=7-11/group). Taken together, our data indicate that Fshb is a Musashi target in the gonadotropes. By deleting MSI from the pituitary gonadotropes, we observe an increase in FSH protein content and serum levels. These Gon-Msi-null female mice have significantly fewer maturing follicles and corpora lutea, which might suggest lower levels of estrogens and progesterone. This, together with the increased GnRHR pituitary protein content, affects LH secretion, leading to a blunted LH surge in the Gon-Msi-null females. Our studies thus reveal a novel translational regulatory mechanism to govern levels of critical reproductive hormones in the pituitary.

Temporal relationships of the LH surge and ovulation to echotexture and power Doppler signals of blood flow in the wall of the preovulatory follicle in heifers

2010 ◽  
Vol 22 (7) ◽  
pp. 1110 ◽  
Author(s):  
M. A. R. Siddiqui ◽  
J. C. Ferreira ◽  
E. L. Gastal ◽  
M. A. Beg ◽  
D. A. Cooper ◽  
...  
Keyword(s):  
Blood Flow ◽  
Power Doppler ◽  
Initial Increase ◽  
Preovulatory Follicle ◽  
Lh Surge ◽  
Follicle Diameter ◽  
Temporal Relationships ◽  
Preovulatory Follicles ◽  
Doppler Signals ◽  
Gonadotrophin Releasing Hormone

Changes in echotexture and blood flow in the wall of preovulatory follicles in heifers were studied in relation to the LH surge and ovulation in gonadotrophin-releasing hormone-induced (n = 7; Experiment 1) and spontaneous (n = 8; Experiment 2) ovulators. Ultrasonographic examinations and blood sampling were performed either every hour (Experiment 1) or every 6 h (Experiment 2). The interval from LH peak to ovulation in induced and spontaneous ovulators was 27.1 ± 0.3 and 34.5 ± 1.5 h, respectively. Follicle diameter did not increase between the LH peak and ovulation. In the induced ovulators, serration of the stratum granulosum was detected in one (14%), two (29%), three (43%) and four (57%) heifers at 4, 3, 2 and 1 h before ovulation, respectively. An initial increase in blood flow (P < 0.001) encompassed the LH peak in both experiments. In the induced ovulators, blood flow increased (P < 0.02) to maximum 3 h after the LH peak, maintained a plateau for 5 h, decreased (P < 0.05) between 9 and 14 h, increased (P < 0.05) again between 19 and 21 h and then decreased (P < 0.01) between 25 and 26 h (1 h before ovulation). The biphasic increase and decrease in blood flow and serration of the granulosum in the wall of the preovulatory follicle in cattle are novel findings.

scholarly journals A mouse model to investigate the impact of testosterone therapy on reproduction in transgender men

2019 ◽  
Vol 34 (10) ◽  
pp. 2009-2017 ◽  
Author(s):  
H M Kinnear ◽  
E S Constance ◽  
A David ◽  
E E Marsh ◽  
V Padmanabhan ◽  
...  
Keyword(s):  
Mouse Model ◽  
Polycystic Ovary ◽  
Case Series ◽  
Corpora Lutea ◽  
Antral Follicles ◽  
Female Mice ◽  
Reproductive Effects ◽  
Vaginal Cytology ◽  
The Impact ◽  
Transgender Men

Abstract STUDY QUESTION Can mice serve as a translational model to investigate the reproductive effects of testosterone (T) therapy commonly used by transgender men? SUMMARY ANSWER T enanthate subcutaneous injections at 0.45 mg twice weekly can be used in the postpubertal C57BL/6N female mouse to investigate the reproductive effects of T therapy given to transgender men. WHAT IS KNOWN ALREADY Most models of T treatment in female mice involve prenatal or prepubertal administration, which are not applicable to transgender men who often begin T therapy after puberty. Studies that have looked at the impact of postpubertal T treatment in female mice have generally not investigated reproductive outcomes. STUDY DESIGN, SIZE, DURATION A total of 20 C57BL/6N female mice were used for this study. Study groups (n = 5 mice per group) included sesame oil vehicle controls and three doses of T enanthate (0.225, 0.45 and 0.90 mg). Mice were injected subcutaneously twice weekly for 6 weeks. PARTICIPANTS/MATERIALS, SETTING, METHODS Daily vaginal cytology was performed prior to initiation of treatment to confirm that all mice were cycling. At 8–9 weeks of age, therapy with subcutaneous T enanthate (0.225, 0.45 or 0.90 mg) or the vehicle control was begun. T therapy continued for 6 weeks, at which point mice were sacrificed and compared to control mice sacrificed during diestrus/metestrus. Data collected included daily vaginal cytology, weekly and terminal reproductive hormone levels, terminal body/organ weights/measurements, ovarian follicular distribution/morphology and corpora lutea counts. MAIN RESULTS AND THE ROLE OF CHANCE Of the mice treated with 0.90 mg T enanthate, two of five mice experienced vaginal prolapse, so this group was excluded from further analysis. T enanthate administration twice weekly at 0.225 or 0.45 mg resulted in cessation of cyclicity and persistent diestrus. One of five mice at the 0.225-mg dose resumed cycling after 2.5 weeks of T therapy. As compared to controls, T-treated mice had sustained elevated T levels and luteinizing hormone (LH) suppression in the terminal blood sample. T-treated mice demonstrated increases in clitoral area and atretic cyst-like late antral follicles (0.45 mg only) as compared to controls. No reduction in primordial, primary, secondary or total antral follicle counts was detected in T-treated mice as compared to controls, and T-treated mice demonstrated an absence of corpora lutea. LIMITATIONS, REASONS FOR CAUTION Mouse models can provide us with relevant key findings for further exploration but may not perfectly mirror human reproductive physiology. WIDER IMPLICATIONS OF THE FINDINGS To our knowledge, this report describes the first mouse model mimicking T therapy given to transgender men that facilitates analysis of reproductive changes. This model allows for future studies comparing duration and reversibility of T-induced changes, on the reproductive and other systems. It supports a role for T therapy in suppressing the hypothalamic–pituitary–gonadal axis in adult female mice as evidenced by LH suppression, persistent diestrus and absence of corpora lutea. The increase in atretic cyst-like late antral follicles aligns with the increased prevalence of polycystic ovary morphology seen in case series of transgender men treated with T therapy. The results also suggest that T therapy does not deplete the ovarian reserve. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the American Society for Reproductive Medicine/Society of Reproductive Endocrinology and Infertility Grant and NIH R01-HD098233 to M.B.M. and University of Michigan Office of Research funding (U058227). H.M.K. was supported by the Career Training in Reproductive Biology and Medical Scientist Training Program T32 NIH Training Grants (T32-HD079342, T32-GM07863) as well as the Cellular and Molecular Biology Program. The University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core is supported by the Eunice Kennedy Shriver NICHD/NIH (NCTRI) Grant P50-HD28934. E.E.M. consults for Allergan. No other authors have competing interests.

scholarly journals KNDy (Kisspeptin/Neurokinin B/Dynorphin) Neurons Are Activated during Both Pulsatile and Surge Secretion of LH in the Ewe

Endocrinology ◽  
2012 ◽  
Vol 153 (11) ◽  
pp. 5406-5414 ◽  
Author(s):  
Christina M. Merkley ◽  
Katrina L. Porter ◽  
Lique M. Coolen ◽  
Stanley M. Hileman ◽  
Heather J. Billings ◽  
...  
Keyword(s):  
Negative Feedback ◽  
Positive Feedback ◽  
Neurokinin B ◽  
Lh Surge ◽  
Short And Long Term ◽  
Positive And Negative Feedback ◽  
Kndy Neurons ◽  
Lh Secretion

Abstract KNDy (kisspeptin/neurokinin B/dynorphin) neurons of the arcuate nucleus (ARC) appear to mediate the negative feedback actions of estradiol and are thought to be key regulators of pulsatile LH secretion. In the ewe, KNDy neurons may also be involved with the positive feedback actions of estradiol (E2) to induce the LH surge, but the role of kisspeptin neurons in the preoptic area (POA) remains unclear. The goal of this study was to identify which population(s) of kisspeptin neurons is (are) activated during the LH surge and in response to the removal of E2-negative feedback, using Fos as an index of neuronal activation. Dual-label immunocytochemistry for kisspeptin and Fos was performed on sections containing the ARC and POA from ewes during the luteal phase of the estrous cycle, or before or after the onset of the LH surge (experiment 1), and from ovary-intact, short-term (24 h) and long-term (&gt;30 d) ovariectomized (OVX) ewes in anestrus (experiment 2). The percentage of kisspeptin neurons expressing Fos in both the ARC and POA was significantly higher during the LH surge. In contrast, the percentage of kisspeptin/Fos colocalization was significantly increased in the ARC, but not POA, after both short- and long-term E2 withdrawal. Thus, POA kisspeptin neurons in the sheep are activated during, and appear to contribute to, E2-positive feedback, whereas ARC kisspeptin (KNDy) neurons are activated during both surge and pulsatile modes of secretion and likely play a role in mediating both positive and negative feedback actions of E2 on GnRH secretion in the ewe.

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