Dynamics of nitric oxide, altered follicular microenvironment, and oocyte quality in women with endometriosis Pravin T. Goud, M.D., Ph.D., Anuradha P.

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Presentation transcript:

Dynamics of nitric oxide, altered follicular microenvironment, and oocyte quality in women with endometriosis Pravin T. Goud, M.D., Ph.D., Anuradha P. Goud, M.Sc., Ph.D., Narendra Joshi, Ph.D., Elizabeth Puscheck, M.S., M.D., Michael P. Diamond, M.D., Husam M. Abu-Soud, Ph.D. Fertility and Sterility Volume 102, Issue 1, Pages 151-159.e5 (July 2014) DOI: 10.1016/j.fertnstert.2014.03.053 Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Bar charts depicting follicular fluid nitrate levels. (A) Follicular fluid nitrate levels were similar between women with (group A) and without (group B) endometriosis. (B) Among women with endometriosis (group A), significantly higher nitrate levels of nitrate were noted among those unable to achieve pregnancy compared with those that did (∗P=.022). (C) On the other hand, the nitrate levels between women without endometriosis were similar among women regardless of success in achieving a pregnancy. (D) Women from groups A and B that conceived with assisted reproductive technology had significantly lower FF nitrate levels compared with those that did not conceive (∗∗P=.0412). Fertility and Sterility 2014 102, 151-159.e5DOI: (10.1016/j.fertnstert.2014.03.053) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Photomicrographs depicting (A–D) follicular granulosa cells and (E–H) in vitro matured metaphase II oocytes from women with and without endometriosis. Granulosa cells were processed for fluorescent staining for chromatin (blue), terminal deoxynucleotide transferase–mediated dUTP nick-end labeling (TUNEL) assay (green), and nitrotyrosine (red). Oocytes were processed for staining for chromatin (blue), microtubules (green) and cortical granules (red). Images represent optical sections obtained with the use of confocal laser scanning microscope with original magnification (×600). Bars = 50 μm; A, B, E, and F at same scale; C, D, G, and H at same scale. (A) Granulosa cells negative for apoptosis in women without endometriosis (group B); (B) granulosa cells with positive TUNEL assay for apoptosis seen in green (arrows). (C) Granulosa cells from women without endometriosis (group B) were negative for nitrotyrosine; (D) whereas those in women with endometriosis were positive for nitrotyrosine (arrows). (E) Intact cortical granules (arrows); (F) premature cortical granule exocytosis in oocytes from women with endometriosis (arrows); the same optical section also depicts disrupted microtubular spindle apparatus (arrowhead). (G) An intact metaphase plate (arrow); (H) a disrupted chromosome metaphase plate (arrow). Fertility and Sterility 2014 102, 151-159.e5DOI: (10.1016/j.fertnstert.2014.03.053) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Design of the present study in the form of a flow chart. Oocytes, follicular fluid, and follicular granulosa cells were collected from women with (group A) and without (group B) endometriosis. Demographics and cycle characteristics were recorded, and mature oocytes were used in the clinical program, whereas their sibling immature oocytes as well as follicular fluid and follicular granulosa cells were subjected to specific tests. ART = assisted reproductive technology; ICSI = intracytoplasmic sperm injection; ZPDT = zona pellucida dissolution time. Fertility and Sterility 2014 102, 151-159.e5DOI: (10.1016/j.fertnstert.2014.03.053) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 2 (A) Bar chart indicates significantly higher zona pellucida dissolution time (ZPDT) among in vitro matured oocytes from group A compared with group B (P=.0038). (B) Scatterplot depicts linear regression analysis. X-axis represents follicular fluid nitrate levels and Y-axis represents ZPDT among in vitro matured oocytes from women in both groups A and B (R2 = 0.5382; P<.0001). (C) Bar chart indicates significantly higher apoptosis among granulosa cells from group A compared with group B (∗P=.013). Fertility and Sterility 2014 102, 151-159.e5DOI: (10.1016/j.fertnstert.2014.03.053) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 3 Bar chart depicting follicular fluid nitrite levels. The levels are noted to be very low with no difference between groups A and B. Fertility and Sterility 2014 102, 151-159.e5DOI: (10.1016/j.fertnstert.2014.03.053) Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions