Screening for embryonic loss during in utero development of mice with a human 1.5 Tesla clinical MRI scanner.

PURPOSE: To establish in utero MRI-scanning of mouse implantation sites in a 1.5 Tesla whole-body human clinical scanner for evaluation of impaired implantation, placental or developmental defects due to genetic alterations. MATERIALS AND METHODS: Pregnant C57Bl/6 wild-type and Cx31-deficient mice revealing placental defects were analyzed in utero using a 1.5 Tesla whole-body clinical scanner in combination with a 3-cm-diameter single loop (slice thickness: 1.2 mm). Imaging of implantation sites was evaluated from 6.5-13.5 dpc and amount of implantation sites and in vivo development was analyzed during the critical phase of placentation from 10.5-13.5 dpc. RESULTS: This method provided high resolution in plane images permitting confident identification of all implantation sites from 6.5 dpc onward. A loss of 60% of Cx31-deficient embryos was demonstrated compared with controls. Repeated anesthesia as well as imaging protocols produced no gross malformations in the surviving mice. CONCLUSION: Using a human clinical MRI scanner high resolution imaging of the entire uterus of the mice and all the embryos inside could be performed. This method is well suited to noninvasively monitor and quantify embryo implantation and to follow this dynamic process in vivo without compromising pregnancy progression and embryonic development.

Regulation of soluble vascular endothelial growth factor receptor (sFlt-1/sVEGFR-1) expression and release in endothelial cells by human follicular fluid and granulosa cells.

BACKGROUND: During the female reproductive cycle, follicular development and corpus luteum formation crucially depend on the fast generation of new blood vessels. The importance of granulosa cells and follicular fluid in controlling this angiogenesis is still not completely understood. Vascular endothelial growth factor (VEGF) produced by granulosa cells and secreted into the follicular fluid plays an essential role in this process. On the other hand, soluble VEGF receptor-1 (sFlt-1) produced by endothelial cells acts as a negative modulator for the bioavailability of VEGF. However, the regulation of sFlt-1 production remains to be determined. METHODS: We analyzed the influence of human follicular fluid obtained from FSH-stimulated women as well as of human granulosa cell conditioned medium on sFlt-1 production in and release from human umbilical vein endothelial cells (HUVEC) in vitro. Soluble Flt-1 gene expression was determined by RT-PCR analysis, amount of sFlt-1-protein was quantified by Sandwich-ELISA. RESULTS: Human follicular fluid as well as granulosa cell-conditioned medium significantly inhibit the production of sFlt-1 by endothelial cells on a posttranscriptional level. Treatment of cultured granulosa cells with either hCG or FSH had not impact on the production of sFlt-1 inhibiting factors. We further present data suggesting that this as yet unknown sFlt-1 regulating factor secreted by granulosa cells is not heat-sensitive, not steroidal, and it is of low molecular mass (< 1000 Da). CONCLUSION: We provide strong support that follicular fluid and granulosa cells control VEGF availability by down regulation of the soluble antagonist sFlt-1 leading to an increase of free, bioactive VEGF for maximal induction of vessel growth in the ovary.