Craig, Z. | Search bioRxiv

bioRxiv2023

Human relevant exposure to di-n-butyl phthalate tampers with the ovarian insulin-like growth factor 1 system and disrupts folliculogenesis in young adult mice

Phthalates are compounds used in consumer and medical products worldwide. Phthalate exposure in women has been demonstrated by detection of phthalate metabolites in their urine and ovarian follicular fluid. High urinary phthalate burden has been associated with reduced ovarian reserve and oocyte retrieval in women undergoing assisted reproduction. Unfortunately, no mechanistic explanation for these associations is available. In short term in vivo and in vitro animal studies modeling human relevant exposures to di-n-butyl phthalate (DBP), we have identified ovarian folliculogenesis as a target for phthalate exposures. In the present study, we investigated whether DBP exposure negatively influences insulin-like growth factor 1 (IGF) signaling in the ovary and disrupts ovarian folliculogenesis. CD-1 female mice were exposed to corn oil (vehicle) or DBP (10 or 100 g/kg/day) for 20-32 days. Ovaries were collected as animals reached the proestrus stage to achieve estrous cycle synchronization. Levels of mRNAs encoding IGF1 and 2 (Igf1 and Igf2), IGF1 receptor (Igf1r), and IGF binding proteins 1-6 (Ifgbp1-6) were measured in whole ovary homogenates. Ovarian follicle counts and immunostaining for phosphorylated IGF1R protein (pIGF1R) were used to evaluate folliculogenesis and IGF1R activation, respectively. DBP exposure, at a realistic dose that some women may experience (100 g/kg/day for 20-32 days), reduced ovarian Igf1 and Igf1r mRNA expression and reduced small ovarian follicle numbers and primary follicle pIGF1R positivity in DBP-treated mice. These findings reveal that DBP tampers with the ovarian IGF1 system and provide molecular insight into how phthalates could influence the ovarian reserve in females.

pharmacology and toxicology↗

bioRxiv2022

A phenomic modeling approach for using chlorophyll-a fluorescence-based measurements on coral photosymbionts: a step towards bio-optical bleaching prediction

We test a newly developed instrument prototype which utilizes time-resolved chlorophyll-a fluorescence techniques and fluctuating light to characterize Symbiodiniaceae functional traits across seven different coral species under cultivation as part of ongoing restoration efforts in the Florida Keys. While traditional chlorophyll-a fluorescence techniques only provide a handful of algal biometrics, the system and protocol we have developed generates > 1000 dynamic measurements in a short ([~]11 min) time frame. Resulting high-content algal biometric data revealed distinct phenotypes, which broadly corresponded to clade-level Symbiodiniaceae designations determined using quantitative PCR. Next, algal biometric data from Acropora cervicornis (10 genotypes) and A. palmata (5 genotypes) coral fragments was correlated with bleaching response metrics collected after a two month-long exposure to high temperature. A network analysis identified 1973 correlations (Spearman R > 0.5) between algal biometrics and various bleaching response metrics. These identified biomarkers of thermal stress were then utilized to train a predictive model, and when tested against the same A. cervicornis and A. palmata coral fragments, yielded high correlation (R = 0.92) with measured thermal response (reductions in absorbance by chlorophyll-a). When applied to all seven coral species, the model ranked fragments dominated by Cladocopium or Breviolum symbionts as more bleaching susceptible than corals harboring thermally tolerant symbionts (Durusdinium). While direct testing of bleaching predictions on novel genotypes is still needed, our device and modeling pipeline may help broaden the scalability of existing approaches for determining thermal tolerance in reef corals. Our instrument prototype and analytical pipeline aligns with recent coral restoration assessments that call for the development of novel tools for improving scalability of coral restoration programs.

ecology↗