Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Feb 4;111(5):1843-8.
doi: 10.1073/pnas.1323416111. Epub 2014 Jan 21.

TAp73 is required for spermatogenesis and the maintenance of male fertility

Satoshi Inoue  1 , Richard Tomasini, Alessandro Rufini, Andrew J Elia, Massimiliano Agostini, Ivano Amelio, Dave Cescon, David Dinsdale, Lily Zhou, Isaac S Harris, Sophie Lac, Jennifer Silvester, Wanda Y Li, Masato Sasaki, Jillian Haight, Anne Brüstle, Andrew Wakeham, Colin McKerlie, Andrea Jurisicova, Gerry Melino, Tak W Mak
Affiliations

TAp73 is required for spermatogenesis and the maintenance of male fertility

Satoshi Inoue et al. Proc Natl Acad Sci U S A. .

Abstract

The generation of viable sperm proceeds through a series of coordinated steps, including germ cell self-renewal, meiotic recombination, and terminal differentiation into functional spermatozoa. The p53 family of transcription factors, including p53, p63, and p73, are critical for many physiological processes, including female fertility, but little is known about their functions in spermatogenesis. Here, we report that deficiency of the TAp73 isoform, but not p53 or ΔNp73, results in male infertility because of severe impairment of spermatogenesis. Mice lacking TAp73 exhibited increased DNA damage and cell death in spermatogonia, disorganized apical ectoplasmic specialization, malformed spermatids, and marked hyperspermia. We demonstrated that TAp73 regulates the mRNA levels of crucial genes involved in germ stem/progenitor cells (CDKN2B), spermatid maturation/spermiogenesis (metalloproteinase and serine proteinase inhibitors), and steroidogenesis (CYP21A2 and progesterone receptor). These alterations of testicular histology and gene expression patterns were specific to TAp73 null mice and not features of mice lacking p53. Our work provides previously unidentified in vivo evidence that TAp73 has a unique role in spermatogenesis that ensures the maintenance of mitotic cells and normal spermiogenesis. These results may have implications for the diagnosis and management of human male infertility.

Keywords: ADAM17; MMP13; Serpin; Timp.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Testes of TAp73 KO mice exhibit altered germ cell numbers, germ cell localization, testicular histology, and ultrastructure. (A and B) The average weights of testes (A) and the number of mature sperm isolated from epididymides (B) were measured in 36-wk-old WT (n = 10) and TAp73 KO (n = 10) mice of the 129Ola background and in 16-wk-old WT (n = 8) and TAp73 KO (n = 8) mice of the C57BL6 (F9) background. Data points are values for individual mice. The horizontal line is the group mean ± SD (*P < 0.02; unpaired Student t test). (C and D) H&E-stained histological sections of epididymides (C) and testes (D) of WT, TAp73 KO [C57BL6 (F9)], and ΔNp73 KO (control) mice of 7 or 11 wk of age. Arrowheads show Leydig cells. (E and F) Electron micrographs of testes from 36-wk-old WT and TAp73 KO mice. (E) Normal basal ES (Basal) of BTB (arrows) surrounding the lower lateral membranes of Sertoli cells in WT and TAp73 KO mice. (F) In contrast to orderly apical ES (apical) in WT testes, TAp73 KO testes show disorganized apical ES populated by malformed spermatids (arrows).
Fig. 2.
Fig. 2.
Fewer spermatids, reduced spermatogonial proliferation, and increased apoptosis and DNA damage in TAp73 KO testes. (A–C) Representative immunohistochemical analysis to detect TAp73 (A), protamine 1 (B), and Ki67+ (C) in testes of 36-wk-old WT and TAp73 KO mice. Arrowheads in A show positive staining in cytoplasm of spermatogonia. (D) Quantitation of the data in C expressed as percentage of Ki67+ cells. Data shown are the means ± SD (n = 5). (E) Representative TUNEL assay detecting apoptosis in testes of 36-wk-old WT (n = 4) and TAp73 KO (n = 5) mice. (F) Quantitation of the data in E expressed as percentage of TUNEL+ cells. Data shown are the means ± SD. (G) Representative immunohistochemical analysis to detect the DNA damage marker γH2AX in testes of 16-wk-old WT (n = 5) and TAp73 KO (n = 5) mice. (H) Quantitation of the data in G expressed as percentage of γH2AX+ cells. Data shown are the mean ± SD. P values were determined according to unpaired Student t test.
Fig. 3.
Fig. 3.
TAp73 plays a unique role in regulating the expression of genes involved in spermatogenesis. (A–C) qPCR determinations of levels of the indicated mRNAs related to markers of differentiated germ cells (A), serine proteinase inhibitors (B), and ADAM, MPPs, and MPP inhibitors (C) in testes of 36-wk-old WT and TAp73 KO mice. Data were analyzed as in A. (D) qPCR determinations of levels of the indicated mRNAs in testes of 18-wk-old WT (n = 3) and p53 KO (n = 3) mice. Data were analyzed as in A.
Fig. 4.
Fig. 4.
MMP13 and ADAM17 are previously unidentified TAp73 target genes. (A) HA tag TAp73β, ΔNp73, and p53 Saos-2 tet-on cells were exposed to 2 μM doxycycline for 16 h, followed by qPCR determinations of the indicated mRNAs. Data shown are the means ± SD (n = 3). P values were determined according to unpaired Student t test. (B) Bioinformatics analysis of human ADAM17 and MMP13 promoters using the MatInspector program has been analyzed for putative p53 binding sites (p53BS). (C) ChIP assay was performed using nuclear extracts from HA-TAp73β–overexpressing Saos-2 cells. Protein–chromatin complexes were immunoprecipitated with anti-HA antibody or control IgG. PCR was performed with primers designed against promoter region predicted or validated p53-binding sites of indicated genes. MDM2- and p21-responsive elements were used as positive controls.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Hess RA, Renato de Franca L. Spermatogenesis and cycle of the seminiferous epithelium. Adv Exp Med Biol. 2008;636:1–15. - PubMed
    1. Cheng CY, Mruk DD. A local autocrine axis in the testes that regulates spermatogenesis. Nat Rev Endocrinol. 2010;6(7):380–395. - PMC - PubMed
    1. Verhoeven G, Willems A, Denolet E, Swinnen JV, De Gendt K. Androgens and spermatogenesis: Lessons from transgenic mouse models. Philos Trans R Soc Lond B Biol Sci. 2010;365(1546):1537–1556. - PMC - PubMed
    1. Melino G, De Laurenzi V, Vousden KH. p73: Friend or foe in tumorigenesis. Nat Rev Cancer. 2002;2(8):605–615. - PubMed
    1. Tomasini R, et al. TAp73 knockout shows genomic instability with infertility and tumor suppressor functions. Genes Dev. 2008;22(19):2677–2691. - PMC - PubMed

Publication types

MeSH terms