ABSTRACT
Maternal-offspring interactions in mammals involve both cooperation and conflict. The fetus has evolved ways to manipulate maternal physiology to enhance placental nutrient transfer, but the mechanisms involved remain unclear. The imprinted Igf2 gene is highly expressed in murine placental endocrine cells. Here, we show that Igf2 deletion in these cells impairs placental endocrine signaling to the mother, without affecting placental morphology. Igf2 controls placental hormone production, including prolactins, and is crucial to establish pregnancy-related insulin resistance and to partition nutrients to the fetus. Consequently, fetuses lacking placental endocrine Igf2 are growth restricted and hypoglycemic. Mechanistically, Igf2 controls protein synthesis and cellular energy homeostasis, actions dependent on the placental endocrine cell type. Igf2 loss also has additional long-lasting effects on offspring metabolism in adulthood. Our study provides compelling evidence for an intrinsic fetal manipulation system operating in placenta that modifies maternal metabolism and fetal resource allocation, with long-term consequences for offspring metabolic health.
Subject(s)
Insulin Resistance , Insulin-Like Growth Factor II , Placenta , Animals , Female , Mice , Pregnancy , Cell Communication , Homeostasis , Hypoglycemic Agents , Insulin-Like Growth Factor II/genetics , Genomic ImprintingABSTRACT
BACKGROUND: RNA is a critical analyte for unambiguous detection of actionable mutations used to guide treatment decisions in oncology. Currently available methods for gene fusion detection include molecular or antibody-based assays, which suffer from either being limited to single-gene targeting, lack of sensitivity, or long turnaround time. The sensitivity and predictive value of next generation sequencing DNA-based assays to detect fusions by sequencing intronic regions is variable, due to the extensive size of introns. The required depth of sequencing and input nucleic acid required can be prohibitive; in addition it is not certain that predicted gene fusions are actually expressed. RESULTS: Herein we describe a method based on pyrophosphorolysis to include detection of gene fusions from RNA, with identical assay steps and conditions to detect somatic mutations in DNA [1], permitting concurrent assessment of DNA and RNA in a single instrument run. CONCLUSION: The limit of detection was under 6 molecules/ 6 µL target volume. The workflow and instrumentation required are akin to PCR assays, and the entire assay from extracted nucleic acid to sample analysis can be completed within a single day.
Subject(s)
Gene Fusion , RNA , High-Throughput Nucleotide Sequencing/methods , Mutation , RNA/genetics , Sequence Analysis, RNAABSTRACT
The results of different human epidemiological datasets provided the impetus to introduce the now commonly accepted theory coined as 'developmental programming', whereby the presence of a stressor during gestation predisposes the growing fetus to develop diseases, such as metabolic dysfunction in later postnatal life. However, in a clinical setting, human lifespan and inaccessibility to tissue for analysis are major limitations to study the molecular mechanisms governing developmental programming. Subsequently, studies using animal models have proved indispensable to the identification of key molecular pathways and epigenetic mechanisms that are dysregulated in metabolic organs of the fetus and adult programmed due to an adverse gestational environment. Rodents such as mice and rats are the most used experimental animals in the study of developmental programming. This review summarises the molecular pathways and epigenetic mechanisms influencing alterations in metabolic tissues of rodent offspring exposed to in utero stress and subsequently programmed for metabolic dysfunction. By comparing molecular mechanisms in a variety of rodent models of in utero stress, we hope to summarise common themes and pathways governing later metabolic dysfunction in the offspring whilst identifying reasons for incongruencies between models so to inform future work. With the continued use and refinement of such models of developmental programming, the scientific community may gain the knowledge required for the targeted treatment of metabolic diseases that have intrauterine origins.
Subject(s)
Disease Models, Animal , Metabolism , Stress, Physiological , Animals , Epigenesis, Genetic , Insulin/metabolism , Liver/embryology , Liver/metabolism , Metabolism/genetics , Rodentia , Stress, Physiological/geneticsABSTRACT
We previously showed that rat, pig, sheep, and red deer oocytes express species-specific ratios of GDF9: BMP15 mRNA (3.7, 0.5, 1.26, and 0.1, respectively), and with the exception of the pig, they are directly correlated to litter size. The purpose of this study was to determine the alternative mechanism that enables pig oocytes to secrete low ratios whilst maintaining a large litter size. Herein, we performed same- and cross-species coincubations of oocytes with granulosa cells (GCs) of rat, pig, sheep, and red deer to compare the proliferation rate, mRNA expression levels of growth factor receptors, and downstream signalling pathways in GCs. A decreased proliferation rate, lower Bmpr1b and Bmpr2 mRNA expression levels, and higher SMAD1/5/8 protein levels were exhibited in rat GCs cocultured with red deer oocytes, compared to all other species. Pig GCs unequivocally expressed GDF9 mRNA, suggesting that, similar to rat GCs, the proliferation of pig GCs is regulated mainly by GDF9, despite lower intraoocyte expression of GDF9 mRNA. In support, a higher basal proliferation, and their ability to proliferate readily when coincubated with red deer oocytes, was observed in pig GCs. In contrast, red deer GC proliferation is likely to be mainly regulated by BMP15 in vivo with only red deer oocytes capable of altering SMAD1/5/8 and pSMAD2/3 levels, while both GDF9 and BMP15 appear important for sheep GC proliferation. In summary, this study strengthens our hypothesis that the ratio of GDF9: BMP15 in the intrafollicular milieu is directly correlated with litter size, and that the GCs of each species have evolved to respond to these unique ratios.