Granular changes in Sertoli cells in children and pubertal patients Manuel Nistal, M.D., Ph.D., Javier Regadera, M.D., Ph.D., Pablo Winitzky, M.Sc., Eva Tejerina, M.D., Hector Chemes, M.D., Ph.D.  Fertility and Sterility  Volume 83, Issue 5, Pages 1489-1499 (May 2005) DOI: 10.1016/j.fertnstert.2004.12.032 Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Granular Sertoli cell (GSC) changes in infantile cryptorchid testes. (A) Numerous seminiferous tubules with complete granular transformation of Sertoli cells (arrows), associated with isolated tubules without granular changes. (B) Decrease of vimentin expression in GSC (arrows) compared with nongranulated cells. (C) Only GSCs express the CD-68 epitope. (D) Alpha-1-antitrypsin is evident in the large lysosomal granules of Sertoli cells. Original magnification: A, B, ×40; C, D, ×200. A, hematoxylin-eosin; B, anti-vimentin; C, anti-CD-68; D, anti-α-1-antitrypsin. Fertility and Sterility 2005 83, 1489-1499DOI: (10.1016/j.fertnstert.2004.12.032) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Granular Sertoli cell (GSC) changes in pubertal patients. (A) Ecstatic tubules contain GSCs (asterisks). Other tubules are normal (star). (B) Granules vary in size, and occupy the entire cytoplasm. (C) Intensely periodic acid-Schiff (PAS) positivity in GSC (asterisk). (D) Lack of inhibin α-subunit expresion in GSC (asterisk). Compare with the intense signal in tubules with spermatogenesis. (E) Sertoli cell–only tubule. Note negativity of inhibin expression in GSC (arrows) in contrast with intense expression in neighboring non-GSCs. (F) Anti-AMH serial section of the same tubule depicted in E. Granular Sertoli cells have a patchy expression of AMH (arrows), some of them with different degrees of positivity and others negative. (G) Granules of Sertoli cells show an intense expression of CD-68 epitope. Original magnification: A, G, ×40; B–D, ×400; E, F, ×600. A, Masson's trichrome stain; B, hematoxylin-eosin; C, PAS stain; D, E, anti-inhibin B; F, anti-AMH; G, anti-CD-68. Fertility and Sterility 2005 83, 1489-1499DOI: (10.1016/j.fertnstert.2004.12.032) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Ultrastructure of human Sertoli cells. (A) Semithin section of a seminiferous tubule in patient 16. Three granular Sertoli cells can be seen. The basal cytoplasm is loaded with dense granules of different shapes and sizes. Sertoli cell nuclei are rounded with prominent nucleoli. (B) Numerous dense heterogeneous granules can be seen surrounding the round nucleus of a Sertoli cell. (C, D) At higher magnification, the heterogeneous nature of Sertoli cell lysosomes is documented. Original magnification: A, ×2,600; B, ×5,200; C, ×11,500; D, ×14,500. Fertility and Sterility 2005 83, 1489-1499DOI: (10.1016/j.fertnstert.2004.12.032) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Ultrastructure of rat Sertoli cells. (A) Basal cytoplasm shows a group of lysosomes depicting various degrees of acid phosphatase activity at stage VIII (before spermiation). (B) After spermiation (stage X), a partially digested residual body inside the Sertoli cell cytoplasm surrounded by lysosomes and a lipid droplet (L). Note the granular heterogeneous structure of the residual body, similar to that seen inside human Sertoli cell granules. Inter-Sertoli junctional complex (arrowheads). (C) At stage XI, a lipid droplet is surrounded by lysosomes with acid phosphatase–positive granules. A partially fused lysosome with acid phosphatase activity can be seen to the right (arrowhead). A, C, acid phosphatase histochemistry, slightly contrasted. Original magnification: A, ×25,600; B, ×8,850; C, ×26,600. Fertility and Sterility 2005 83, 1489-1499DOI: (10.1016/j.fertnstert.2004.12.032) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions