Neural Circuits and Hormonal Mechanisms Underlying Female Reproduction

‘Homeostasis’ is the physiological process of maintaining a stable equilibrium, operating within optimal limits to promote health and prevent illness. Deviations from homeostasis can have adverse consequences for one’s health. However, physiological and behavioral needs vary markedly over the course of the day, necessitating that biological systems are adjusted correspondingly. Likewise, physiological needs change over the female reproductive cycle, with ovulation, pregnancy and fetal development, and parturition requiring specifically-timed patterns of hormone secretion regulated by the circadian system. Disruption to homeostasis, either by circadian misalignment or by exposure to stress, has marked, negative consequences for female reproductive health, including ovulation, pregnancy success and maintenance, and offspring development. The overarching goal of this dissertation research is to understand the neural and hormonal mechanisms underlying reproductive health and how these mechanisms are affected by disruption to homeostasis through circadian misalignment and stress. This dissertation considers two aspects of the female reproductive cycle, the ovulatory cycle and pregnancy. Despite a significant volume of knowledge detailing the circadian regulation of ovulation and the negative impact of stress of reproductive health, the mechanisms underlying these events remain poorly understood. Chapter 2 investigates the time-dependent sensitivity of the reproductive axis to the inhibitory neuropeptide RFamide-related peptide-3 (RFRP-3) in the control of the neuroendocrine events required for ovulation. Chapters 3-5 explore how stress hormones act on the brain, ovaries, and placenta to compromise pregnancy success and fetal development. These studies inform our understanding of the complex neural and endocrine networks regulating environmental and physiological processes involved in promoting female reproductive health

Circadian control of neuroendocrine function: implications for health and disease

The circadian timing system orchestrates daily rhythms in physiology and behavior via the suprachiasmatic nucleus (SCN), the master brain clock. Because endocrine secretions have far-reaching influence on the brain and periphery, circadian regulation of hormones is essential for normal functioning and disruptions to circadian timing (e.g., irregular sleep patterns, limited exposure to sunlight, jet lag, nighttime light exposure) have detrimental health consequences. Herein, we provide an overview of circadian timing in three major endocrine axes, the hypothalamo-pituitary-gonadal (HPG), hypothalamo-pituitary-adrenal (HPA) and hypothalamo-pituitary-thyroid (HPT) axes, and then consider the negative health consequences of circadian disruptions in each of these systems. For example, disruptions to HPG axis circadian timing lead to a host of negative reproductive outcomes such as irregular menstrual cycles, low sperm density and increased rates of miscarriages and infertility. Dysregulation of HPA axis timing is associated with obesity and metabolic disease, whereas disruptions to the HPT axis are associated with dysregulated metabolic gene rhythms in the heart. Together, this overview underscores the significance of circadian endocrine rhythms in normal health and disease prevention

Time‐of‐day‐dependent sensitivity of the reproductive axis to RFamide‐related peptide‐3 inhibition in female Syrian hamsters

In spontaneously ovulating rodent species, the timing of the luteinising hormone (LH) surge is controlled by the master circadian pacemaker in the suprachiasmatic nucleus (SCN). The SCN initiates the LH surge via the coordinated control of two opposing neuropeptidergic systems that lie upstream of the gonadotrophin-releasing hormone (GnRH) neuronal system: the stimulatory peptide, kisspeptin, and the inhibitory peptide, RFamide-related peptide-3 (RFRP-3; the mammalian orthologue of avian gonadotrophin-inhibitory hormone [GnIH]). We have previously shown that the GnRH system exhibits time-dependent sensitivity to kisspeptin stimulation, further contributing to the precise timing of the LH surge. To examine whether this time-dependent sensitivity of the GnRH system is unique to kisspeptin or a more common mechanism of regulatory control, we explored daily changes in the response of the GnRH system to RFRP-3 inhibition. Female Syrian hamsters were ovariectomised to eliminate oestradiol (E2 )-negative-feedback and RFRP-3 or saline was centrally administered in the morning or late afternoon. LH concentrations and Lhβ mRNA expression did not differ between morning RFRP-3-and saline-treated groups, although they were markedly suppressed by RFRP-3 administration in the afternoon. However, RFRP-3 inhibition of circulating LH at the time of the surge does not appear to act via the GnRH system because no differences in medial preoptic area Gnrh or RFRP-3 receptor Gpr147 mRNA expression were observed. Rather, RFRP-3 suppressed arcuate nucleus Kiss1 mRNA expression and potentially impacted pituitary gonadotrophs directly. Taken together, these findings reveal time-dependent responsiveness of the reproductive axis to RFRP-3 inhibition, possibly via variation in the sensitivity of arcuate nucleus kisspeptin neurones to this neuropeptide

Hepatitis B virus downregulates vitamin D receptor levels in hepatoma cell lines, thereby preventing vitamin D-dependent inhibition of viral transcription and production

Abstract Background Vitamin D is a key immune-modulator that plays a role in the innate and adaptive immune systems. Certain pathogens impair the immune defense by downregulating the vitamin D receptor (VDR) pathway. Low serum levels of vitamin D are associated with increased hepatitis B virus (HBV) replication. Our study aimed to assess the in-vitro relationship between HBV production and Vitamin D signaling pathway and to explore the associated mechanism(s). Methods HBV transcription and replication was evaluated by qRT-PCR of the HBV-RNA and covalently closed circular DNA (cccDNA). Furthermore, we have transfected the 1.3 X HBV-Luc plasmid to the cells and measured the Luciferase activity using Luminometer. Vitamin D signaling pathway activation was evaluated by measuring the expression levels of VDR, CYP24A1, Tumor necrosis factor α (TNFα) and cathelicidin (CAMP) by qRT-PCR. All assays were performed on HepG2.2.15, HepG2, and HepAD38 cells treated with or without Vitamin D active metabolite: calcitriol. Results Calcitriol did not suppress HBV transcription, cccDNA expression or HBV RNA levels in HepG2.2.15 cells. However, VDR transcript levels in HepG2.215 cells were significantly lower compared to HepG2 cells. Similar results were obtained in HepAD38 cell where VDR expression was down-regulated when HBV transcript level was up-regulated. In addition, calcitriol induced VDR-associated signaling, resulting in upregulation of CYP24A1, TNFα and CAMP expression level in HepG2 cells but not in the HepG2.2.15 cells. Conclusions These findings indicate that VDR expression is downregulated in HBV-transfected cells, thereby preventing vitamin D from inhibiting transcription and translation of HBV in vitro. HBV might use this mechanism to avoid the immunological defense system by affecting both TNFα and CAMP signaling pathways

Predicting labor onset relative to the estimated date of delivery using smart ring physiological data

Abstract The transition from pregnancy into parturition is physiologically directed by maternal, fetal and placental tissues. We hypothesize that these processes may be reflected in maternal physiological metrics. We enrolled pregnant participants in the third-trimester (n = 118) to study continuously worn smart ring devices monitoring heart rate, heart rate variability, skin temperature, sleep and physical activity from negative temperature coefficient, 3-D accelerometer and infrared photoplethysmography sensors. Weekly surveys assessed labor symptoms, pain, fatigue and mood. We estimated the association between each metric, gestational age, and the likelihood of a participant’s labor beginning prior to (versus after) the clinical estimated delivery date (EDD) of 40.0 weeks with mixed effects regression. A boosted random forest was trained on the physiological metrics to predict pregnancies that naturally passed the EDD versus undergoing onset of labor prior to the EDD. Here we report that many raw sleep, activity, pain, fatigue and labor symptom metrics are correlated with gestational age. As gestational age advances, pregnant individuals have lower resting heart rate 0.357 beats/minute/week, 0.84 higher heart rate variability (milliseconds) and shorter durations of physical activity and sleep. Further, random forest predictions determine pregnancies that would pass the EDD with accuracy of 0.71 (area under the receiver operating curve). Self-reported symptoms of labor correlate with increased gestational age and not with the timing of labor (relative to EDD) or onset of spontaneous labor. The use of maternal smart ring-derived physiological data in the third-trimester may improve prediction of the natural duration of pregnancy relative to the EDD