ABSTRACT
The importance of placental circulation is exemplified by the correlation of placental size and blood flow with fetal weight and survival during normal and compromised human pregnancies in such conditions as preeclampsia and intrauterine growth restriction (IUGR). Using noninvasive magnetic resonance imaging, we evaluated the role of PKBalpha/AKT1, a major mediator of angiogenesis, on placental vascular function. PKBalpha/AKT1 deficiency reduced maternal blood volume fraction without affecting the integrity of the fetomaternal blood barrier. In addition to angiogenesis, PKBalpha/AKT1 regulates additional processes related to survival and growth. In accordance with reports in adult mice, we demonstrated a role for PKBalpha/AKT1 in regulating chondrocyte organization in fetal long bones. Using tetraploid complementation experiments with PKBalpha/AKT1-expressing placentas, we found that although placental PKBalpha/AKT1 restored fetal survival, fetal PKBalpha/AKT1 regulated fetal size, because tetraploid complementation did not prevent intrauterine growth retardation. Histological examination of rescued fetuses showed reduced liver blood vessel and renal glomeruli capillary density in PKBalpha/Akt1 null fetuses, both of which were restored by tetraploid complementation. However, bone development was still impaired in tetraploid-rescued PKBalpha/Akt1 null fetuses. Although PKBalpha/AKT1-expressing placentas restored chondrocyte cell number in the hypertrophic layer of humeri, fetal PKBalpha/AKT1 was found to be necessary for chondrocyte columnar organization. Remarkably, a dose-dependent phenotype was exhibited for PKBalpha/AKT1 when examining PKBalpha/Akt1 heterozygous fetuses as well as those complemented by tetraploid placentas. The differential role of PKBalpha/AKT1 on mouse fetal survival and growth may shed light on its roles in human IUGR.
Subject(s)
Body Size/genetics , Fetal Viability/genetics , Fetus/physiology , Placenta/metabolism , Proto-Oncogene Proteins c-akt/physiology , Animals , Embryo, Mammalian , Female , Fetal Growth Retardation/genetics , Fetal Growth Retardation/metabolism , Fetal Growth Retardation/pathology , Fetal Viability/physiology , Fetus/metabolism , Gestational Age , Male , Mice , Mice, Inbred C57BL , Mice, Inbred ICR , Mice, Knockout , Pregnancy , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-akt/metabolismABSTRACT
PURPOSE: The purpose of this study was to develop a tool for functional phenotyping of the maternal circulation in the mouse placenta. PROCEDURES: In utero macromolecular dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was performed on embryonic day 10.5 (E10.5), E13.5, and E18.5. Fluorescence analysis was also used for validation of the results. RESULTS: The initial rate of contrast enhancement revealed an increased maternal blood volume fraction as the pregnancy progressed. Serial imaging of E10.5 and E13.5 placentas revealed a loss of contrast enhancement due to phagocytic uptake. A key application of macromolecular DCE-MRI would be to follow mouse pregnancies during fetal and placental manipulation including embryo transfer, tetraploid complementation, and fetal resorptions. We were able to resolve strain differences in ICR outbred mice carrying both ICR and C57Bl/6J embryos and to differentiate in utero resorptions from functional placentas. CONCLUSIONS: Our results highlight the importance of the functional in utero analysis of placental vascularization in physiological phenotyping of transgenic mice and suggest MRI, particularly macromolecular DCE-MRI, as a non-invasive tool for the analysis of the placenta.
