Dietary glycine does not affect physiological angiogenesis and reproductive function, but inhibits apoptosis in endometrial and ovarian tissue by down-regulation of nuclear factor-κB  Matthias W. Laschke, M.D., Christine Schwender, Claudia Scheuer, Ph.D., Brigitte Vollmar, M.D., Michael D. Menger, M.D.  Fertility and Sterility  Volume 90, Issue 4, Pages 1460-1469 (October 2008) DOI: 10.1016/j.fertnstert.2007.08.047 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Intravital fluorescence microscopy of endometrial fragments (A, B; borders marked by arrows) and ovarian follicles (C, D; borders marked by arrows) at day 10 after transplantation into the dorsal skinfold chambers of glycine-treated (A, C) and control Syrian golden hamsters (B, D). Note that all grafts exhibit complete, newly developed microvascular networks with a glomerulum-like angioarchitecture. Blue light epi-illumination with contrast enhancement by 5% fluorescein isothiocyanate (FITC)-labeled IV dextran 150,000. Scale bars, 280 μm. Vascularized area (%) (E, F) and microvessel density (cm/cm2) (G, H) of endometrial fragments (E, G) and ovarian follicles (F, H) after transplantation into the dorsal skinfold chambers of control (open circles) and glycine-treated (closed circles) Syrian golden hamsters, as assessed by intravital fluorescence microscopy and computer-assisted image analysis. Means ± SEM. aP<.05 vs. day 0 within each individual group; bP<.05 vs. days 0 and 2 within each individual group; cP<.05 vs. days 0, 2, and 4 within each individual group. Fertility and Sterility 2008 90, 1460-1469DOI: (10.1016/j.fertnstert.2007.08.047) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Hematoxylin and eosin-stained cross-sections of endometrial fragments (A, B) and ovarian follicles (C, D) at day 14 after transplantation onto the striated muscle tissue (arrows) within the dorsal skinfold chamber of glycine-treated (A, C) and control Syrian golden hamsters (B, D). In both groups, endometrial grafts are characterized by cyst-like dilated endometrial glands (asterisks) with an intact glandular epithelium surrounded by a vascularized endometrial stroma comparable to endometriotic lesions (A, B). Transplanted ovarian follicles of both groups are characterized by several layers of granulosa cells (GC) surrounding the oocyte (asterisks) and a well-developed vascularized theca interna, reflecting the stage of late secondary follicles (C, D). Scale bars, 120 μm. Atretic follicles (%) (E) in eutopic ovaries of control and glycine-treated Syrian golden hamsters, as assessed by immunohistochemical detection of caspase-3 in GC undergoing apoptotic cell death. Means ± SEM. Representative images of an atretic (F) and a normal follicle (G) within the ovary of a control hamster. Immunohistochemical staining of caspase-3 displays many apoptotic GC (arrows) surrounding the oocyte (asterisk) of the atretic follicle, whereas the normal follicle is characterized by a lack of apoptotic cell death. Scale bars, 40 μm. Fertility and Sterility 2008 90, 1460-1469DOI: (10.1016/j.fertnstert.2007.08.047) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Western blot analysis of proliferating cell nuclear antigen (PCNA), cleaved caspase-3, p53, and NF-κB protein expression (optical density × mm2) of eutopic endometrium (A) and ovary (B), which were isolated from glycine-treated (black bars) and control Syrian golden hamsters (white bars). Means ± SEM. ∗P<.05 vs. control. Fertility and Sterility 2008 90, 1460-1469DOI: (10.1016/j.fertnstert.2007.08.047) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions