FIGURE SUMMARY
Title

Zebrafish androgen receptor is required for spermatogenesis and maintenance of ovarian function

Authors
Yu, G., Zhang, D., Liu, W., Wang, J., Liu, X., Zhou, C., Gui, J., Xiao, W.
Source
Full text @ Oncotarget

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Observed In:
Stage: Adult

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ar-null male zebrafish have female secondary sex characteristics. (A-C) Wildtype (ar+/+) and heterozygous (ar+/-) male adult zebrafish have deep yellow pigmentation of the anal fin and breeding tubercles (BT) in the pectoral fin (red arrows); ar-null male zebrafish (ar -/-; ar ihb1225/ihb1225) had light yellow pigmentation of the anal fin and no pectoral BTs; ar-null male zebrafish (ar-/-; ar ihb1225/ihb1225) also had a female-like appearance and a larger abdomen. Mpf, month post fertilization.

Targeted disruption of ar in zebrafish causes male infertility due to defective spermatogenesis. (A-C) Gross appearance of testes from wildtype (ar +/+), heterozygous (ar +/-) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish. Testes of ar -/- male zebrafish was smaller and more transparent compared with that in their ar +/+ or ar +/- male siblings. (D) Gonadosomatic index (GSI) in ar+/+ and ar-/- male zebrafish. (E) Sperm motility evaluation under a dark-phase microscope. (F, G) Histology of testes from wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 3 mpf. Compared with wildtype sibling testes, spermatogenesis in ar -/- testes was delayed as indicated by more spermatogonia (SG), fewer spermatocytes (SC) and fewer spermatozoa (SZ); the spermatogenetic cysts were smaller; and spermatogenesis was arrested at the second meiosis. (H, I) Toluidine blue staining testes from wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 3 mpf. Compared with wildtype sibling testes, the size of spermatogonia (SG) was bigger and the number of Sertoli cells was increased in ar -/- testes. Leydig cells (LC) and lumen are indicated. Dpf, days post fertilization; Mpf, months post fertilization.

Proliferating germ cells were reduced, but apoptotic germ cells increased in ar-null zebrafish testes. (A, B) Immunofluorescent staining with anti-vasa antibody identified different types of spermatogenic cells in ar -/- testes and wildtype sibling (ar +/+) testes at 70 dpf or 4.5 mpf, respectively. SG, spermatogonia; SZ, spermatozoa; PSP, primary spermatocyte; SSP, secondary spermatocyte; SPD, spermatid. (C) Immunohistochemistry with anti-cyclin D1 antibody indicated that proliferating germ cells were reduced in ar -/- testes compared with wildtype (ar+/+) siblings at 4.5 mpf. (D) TUNEL assay indicated increased apoptosis of germ cells in ar -/- testes compared with wildtype (ar +/+) siblings at 4.5 mpf. Mpf, months post fertilization. (E, F) Expression of ccnd2a (cyclin d2a) was down-regulated, but expression of gsdf was up-regulated in ar-/- (ar ihb1225/ihb1225) testes compared with wildtype (ar +/+) sibling testes (4.5 mpf, n=3, respectively).

Targeted disruption of ar in zebrafish causes premature ovarian failure during growth. (A-C) Gross appearance and ovaries from wildtype (ar +/+), heterozygous (ar +/-) and homozygous (ar -/-) (ar ihb1225/ihb1225) female zebrafish. At 4.5 mpf, the gross appearance of ar -/- (ar ihb1225/ihb1225) female zebrafish was indistinguishable from wildtype siblings (ar +/+) or their heterozygous sibling (ar +/-), but the ovaries of ar -/- female were smaller with fewer eggs and became transparent compared with wildtype sibling (ar +/+) or heterozygous siblings (ar +/-). (D) Gonadosomatic index (GSI) in ar +/+ and ar -/- female zebrafish. (E) Fertilization rate of eggs from wildtype (ar +/+) and homozygous (ar -/-) female zebrafish at 4mpf. (F, G) Histology of ovaries from wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 1.5 mpf. No obvious difference between ar -/- and ar +/+ ovaries. OC, Oocyte. (H, I) Histology of ovary from the wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 70 dpf. Ovaries of wildtype (ar +/+), most oocytes developed to stage III and IV; but more ovaries of homozygous (ar -/-) (M1) developed to stage II or I, some developed to stage III and IV. (J, K) Histology of ovaries from wildtype (ar +/+) and homozygous (ar -/-) zebrafish at 4 mpf. In ovaries of wildtype (ar +/+), most oocytes developed to stage IV, but for ovaries of homozygous (ar -/-), most oocytes developed to stage II and partial degeneration occurred. (L, M) Histology of ovaries from wildtype (ar+/+) and homozygous (ar -/-) zebrafish at 4 mpf. In ovaries of homozygous (ar -/-), follicles atresia and degeneration occurred. (N, O) Histology of ovaries from wildtype (ar +/+) and homozygous (ar -/-) zebrafish at 5 mpf. In ovaries of homozygous (ar -/-), no follicles were observed and severe degeneration occurred (30%, n = 30). (P) lhcgr (luteinizing homone/choriogonadotropin receptor) expression in ovaries of wildtype (ar+/+) and homozygous (ar-/-) zebrafish at 4.5 mpf. (Q) foxl2 (forkhead box L2) expression in ovaries of wildtype (ar +/+) and homozygous (ar -/-) zebrafish at 4.5 mpf. (R) cyp11a1 (p450 side chain cleavage enzyme) expression in ovaries of wildtype (ar+/+) and homozygous (ar -/-) zebrafish at 4.5 mpf. (S) kitlga expression in ovaries of wildtype (ar+/+) and homozygous (ar -/-) zebrafish at 4.5 mpf injected with or without DHT (100 nM, 10μl for each). Dpf, days post fertilization; Mpf, months post fertilization.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Observed In:
Stage: Adult

General appearance of testis from wildtype (ar +/+), heterozygous (ar +/-) and homozygous (ar -/-) zebrafish at 4.5 mpf. (A) ar +/+. (B) ar +/-. (C) ar -/-. Mpf, months post fertilization.

PHENOTYPE:
Fish:
Observed In:
Stage: Adult

(A, B) At 3 or 4.5 mpf, ovary eggs were fewer in ar -/- females compared to wildtype siblings (ar +/+) (n=5 groups, which were measured everyday for 2 weeks). (C, D) Immunofluorescent staining using anti-vasa antibody identified different stages of oogenesis in ovaries from wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 4.5 mpf. I, stage I oocyte; II, stage II; III, stage III; IV, stage IV. Mpf, months post fertilization.

Histology of testis from wildtype (ar +/+), and homozygous (ar -/-) (the mutant 1) zebrafish at 1.5 mpf, 70dpf and 5 mpf. (A, B) Histology of testes from the wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 1.5 mpf. No obvious difference between ar -/- and wildtype sibling (ar +/+) testes. SGC, spermatogonial cysts (indicated with red arrows). (C, D) Histology of testes from the wildtype (ar +/+) and homozygous (ar -/-) (ar ihb1225/ihb1225) zebrafish at 70 dpf. Compared with wildtype sibling testes, spermatogenesis of ar-/- testes was delayed as indicated by more spermatogonia (SG) and fewer spermatocytes (SC), but no spermatozoa (SZ). (E, F) Histology of testes from the wildtype (ar +/+) and homozygous (ar -/-) zebrafish at 6 mpf. ar -/- testes were degenerated and loose; but spermatozoa were filled in wildtype sibling (ar +/+) testes. Dpf, days post fertilization; Mpf, months post fertilization.

Histology of testis and ovary from wildtype (ar +/+), heterozygous (ar +/-) and homozygous (ar -/-) (the mutant 2) zebrafish at 3 mpf. (A) Testis. (B) Ovary. Mpf, months post fertilization.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Oncotarget