Noninvasive assessment of ectopic uterine tissue development in rats using magnetic resonance imaging  Stephen C. Lenhard, M.S., Robin E. Haimbach, B.S., Anthony C. Sulpizio, B.S., David P. Brooks, Ph.D., Jeffrey D. Bray, Ph.D., Beat M. Jucker, Ph.D.  Fertility and Sterility  Volume 88, Issue 4, Pages 1058-1064 (October 2007) DOI: 10.1016/j.fertnstert.2006.11.164 Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Estrous pattern displayed by low activity periods. Nocturnal low activity periods (<21 activity counts/15-min period) were collected for 1 month and coincide with the 4-day estrous cycle. Each trough corresponds to the night before a day of estrus. Lenhard. MRI of implanted uterine tissue in rat. Fertil Steril 2007. Fertility and Sterility 2007 88, 1058-1064DOI: (10.1016/j.fertnstert.2006.11.164) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Early ectopic uterine tissue (EUT) development. The magnetic resonance images (MRI) of early EUT growth in the same rat from day of transplantation surgery (day 0) through to day 4 after surgery are shown (A). A magnified view of the EUT attachment to the peritoneal wall may be observed in the figure inset. Mean daily EUT volume increased 54%, 117%, 220%, and 279%, respectively, from day 1 through day 4 after surgery versus day 0 (B). Data are presented as mean ± SEM. ∗P<.001 vs. day 0. Lenhard. MRI of implanted uterine tissue in rat. Fertil Steril 2007. Fertility and Sterility 2007 88, 1058-1064DOI: (10.1016/j.fertnstert.2006.11.164) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effect of ovariectomy on EUT volume. (A) MRIs of the same representative rat's preovariectomy and 2 weeks postovariectomy EUT. The EUT volumes decreased after ovariectomy and achieved a plateau volume at approximately 6 weeks after ovariectomy (n = 6) (B). Data are presented as mean ± SEM. ∗P<.05, ∗∗P<.01, †P<.001 vs. baseline. Lenhard. MRI of implanted uterine tissue in rat. Fertil Steril 2007. Fertility and Sterility 2007 88, 1058-1064DOI: (10.1016/j.fertnstert.2006.11.164) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Effect of Antide on EUT volume with or without hormone supplementation. The MRIs were acquired of the transplanted EUT at baseline (n = 12). Antide (GnRH antagonist) was administered every 3 days for 12 days at 2 mg/kg/day. After 12 days on Antide, rats were divided into two groups to receive hormone supplementation with either 17β-E2 (0.08 mg/kg/day, n = 6) or P (4.0 mg/kg/day, n = 6). Antide treatment significantly reduced EUT volumes, whereas 17β- E2 supplementation significantly increased EUT volumes after 4 and 8 days of treatment. Progesterone supplementation did not affect EUT volumes at either time point. Data are presented as mean ± SEM. ∗P<.0001 vs. baseline; ∗∗P<.05 vs. Antide. Lenhard. MRI of implanted uterine tissue in rat. Fertil Steril 2007. Fertility and Sterility 2007 88, 1058-1064DOI: (10.1016/j.fertnstert.2006.11.164) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 5 The MRI assessment of transplanted EUT perfusion characteristics. MRIs of an established EUT (∼5 weeks after transplantation) pre (A) and ∼2 minutes after bolus injection of Gd-DTPA contrast agent (B). Early EUT growth and perfusion was monitored by imaging immediately after bolus injections of Gd-DTPA on the day of EUT transplantation (day 0) (C) and at 3 days after transplantation (D). Lenhard. MRI of implanted uterine tissue in rat. Fertil Steril 2007. Fertility and Sterility 2007 88, 1058-1064DOI: (10.1016/j.fertnstert.2006.11.164) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions