Sexing of postimplantation rat embryos in stored twodimensional electrophoresis (2-DE) samples by polymerase chain reaction (PCR) of an Sry sequence

Miyajima, Atsuko; Sunouchi, Momoko; Mitsunaga, Katsuyoshi; Yamakoshi, Yoko; Nakazawa, Ken; Usami, Makoto

doi:10.2131/jts.34.681

Cited by 19 publications

(11 citation statements)

References 13 publications

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“…Amplification of both genes was carried out in the same reaction due to limited DNA content. Primers for Sry (317 bp) and B-Actin (220 bp) were derived from Miyajima (2009), ref [39] for use in the rat (see Additional file 1 for primer sequences and PCR conditions). A 2% agarose gel was run for 35 min at 100 V to separate the bands.…”

Section: Methodsmentioning

confidence: 99%

Sex differences in rat placental development: from pre-implantation to late gestation

et al. 2017

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BackgroundA male fetus is suggested to be more susceptible to in utero and birth complications. This may be due in part to altered morphology or function of the XY placenta. We hypothesised that sexual dimorphism begins at the blastocyst stage with sex differences in the progenitor trophectoderm (TE) and its derived trophoblast lineages, as these cells populate the majority of cell types within the placenta. We investigated sex-specific differences in cell allocation in the pre-implantation embryo and further characterised growth and gene expression of the placental compartments from the early stages of the definitive placenta through to late gestation.MethodsNaturally mated Sprague Dawley dams were used to collect blastocysts at embryonic day (E) 5 to characterise cell allocation; total, TE, and inner cell mass (ICM), and differentiation to downstream trophoblast cell types. Placental tissues were collected at E13, E15, and E20 to characterise volumes of placental compartments, and sex-specific gene expression profiles.ResultsPre-implantation embryos showed no sex differences in cell allocation (total, TE and ICM) or early trophoblast differentiation, assessed by outgrowth area, number and ploidy of trophoblasts and P-TGCs, and expression of markers of trophoblast stem cell state or differentiation. Whilst no changes in placental structures were found in the immature E13 placenta, the definitive E15 placenta from female fetuses had reduced labyrinthine volume, fetal and maternal blood space volume, as well as fetal blood space surface area, when compared to placentas from males. No differences between the sexes in labyrinth trophoblast volume or interhaemal membrane thickness were found. By E20 these sex-specific placental differences were no longer present, but female fetuses weighed less than their male counterparts. Coupled with expression profiles from E13 and E15 placental samples may suggest a developmental delay in placental differentiation.ConclusionsAlthough there were no overt differences in blastocyst cell number or early placental development, reduced growth of the female labyrinth in mid gestation is likely to contribute to lower fetal weight in females at E20. These data suggest sex differences in fetal growth trajectories are due at least in part, to differences in placenta growth.Electronic supplementary materialThe online version of this article (doi:10.1186/s13293-017-0138-6) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Sex differences in rat placental development: from pre-implantation to late gestation

et al. 2017

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“…Because the sex of newborns rats is often ambiguous, genetic screening was used to confirm sex as described elsewhere 29 . Briefly, DNA was isolated from tissue samples, and Sex-determining region Y ( Sry , male-specific) and beta actin (autosomal) gene sequences were amplified by polymerase chain reaction (PCR) using Taq DNA polymerase (G-Biosciences, St. Louis, MO) and primers obtained from Eurofins MWG Operon (Huntsville, AL).…”

Section: Methodsmentioning

confidence: 99%

Isoflurane exposure in newborn rats induces long-term cognitive dysfunction in males but not females

et al. 2014

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Volatile anesthetics are used widely for achieving a state of unconsciousness, yet these agents are incompletely understood in their mechanisms of action and effects on neural development. There is mounting evidence that children exposed to anesthetic agents sustain lasting effects on learning and memory. The explanation for these behavioral changes remains elusive, although acute neuronal death after anesthesia is commonly believed to be a principal cause. Rodent models have shown that isoflurane exposure in newborns induces acute neuroapoptosis and long-term cognitive impairment. However, the assessment of predisposing factors is lacking. We investigated the role of sex by delivering isoflurane to postnatal day (P)7 male and female Sprague Dawley rats for 4 hours. Brain cell death was assessed 12 h later using FluoroJade C staining in the thalamus, CA1-3 regions of hippocampus, and dentate gyrus. Behavior was assessed separately using a series of object recognition tasks and a test of social memory beginning at P38. We found that isoflurane exposure significantly increased neuronal death in each brain region with no difference between sexes. Behavioral outcome was also equivalent in simple novel object recognition. However, only males were impaired in the recognition of objects in different locations and contexts. Males also exhibited deficient social memory while females were intact. The profound behavioral impairment in males relative to females, in spite of comparable cell death, suggests that males are more susceptible to long-term cognitive effects and this outcome may not be exclusively attributed to neuronal death.

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“…The PCR machine was set to 35 cycles, step 1: 95˚C (1 min), step 2: 52˚C (1 min) step 3: 72˚C (1 min), final extension: 72˚C (5 min) and a final hold at 10˚C. The product was loaded onto a 1.8% agarose gel and sex was determined by presence of Sry band (Figure 2A; adapted from Miyajima et al, 2009).…”

Section: Extraction Of Genomic Dna and Polymerase Chain Reactionmentioning

confidence: 99%

Environmental influences on placental programming and offspring outcomes following maternal immune activation

Estevez

Rondón-Ortiz

Nguyen

et al. 2020

Brain, Behavior, and Immunity

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Adverse experiences during pregnancy induce placental programming, affecting the fetus and its developmental trajectory. However, the influence of 'positive' maternal experiences on the placenta and fetus remain unclear. In animal models of early life stress, environmental enrichment (EE) has ameliorated and even prevented associated impairments in brain and behavior. Here, using a maternal immune activation (MIA) model in rats, we test whether EE attenuates maternal, placental and/or fetal responses to an inflammatory challenge, thereby offering a mechanism by which fetal programming may be prevented. Moreover, we evaluate life-long EE exposure on

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Sexing of postimplantation rat embryos in stored twodimensional electrophoresis (2-DE) samples by polymerase chain reaction (PCR) of an Sry sequence

Cited by 19 publications

References 13 publications

Sex differences in rat placental development: from pre-implantation to late gestation

Sex differences in rat placental development: from pre-implantation to late gestation

Isoflurane exposure in newborn rats induces long-term cognitive dysfunction in males but not females

Environmental influences on placental programming and offspring outcomes following maternal immune activation

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