Fig 1.
Expression of PKnox1 in postnatal testes.
(A) Expression of PKnox1 and β-actin (control) mRNA transcripts in testes at postnatal (P) days 6, 14, 35 and 6 months obtained from wild-type mice and 12-week-old W/Wv mice. (B-E) Immunohistochemical analysis to localize PKnox1-expressing cells in the adult testis. Serial tissue sections of the testes from 8-week-old wild-type mice were stained with a PKnox1-specific antibody, in combination with anti-c-Kit (B, C) or PLZF antibodies (D, E). Inserts indicate cells co-expressing both Prep1 and c-Kit or PLZF. Asterisks indicate haploid cells positive for PKnox1. Data are representative of 3 independent experiments. Scale bar, 50 μm.
Fig 2.
Loss of PKnox1 in the adult testis causes defective spermatogenesis.
The size (A) and weight (B) of adult testes from 12-week-old PKnox1-CKO and control mice 3 weeks after induction of PKnox1 deletion. Bars are the mean and standard deviation of the weight of testes. (n = 4 for each genotype). Tissue sections from control and CKO epididymis stained with H&E (C, D) and testes stained with H&E (E, F), TUNEL (G, H) and anti-SCP3 antibody with DAPI (I, J). Data are representative of 4 independent experiments. Scale bars, 50 μm.
Fig 3.
Germ cell-specific PKnox1 loss causes defective spermatogenesis.
The size (A) and weight (B) of testes from 12-week-old PKnox1-GKO and control mice. Bars are the mean and standard deviation of the weight of testes. (n = 4 for each genotype). Tissue sections from control and GKO epididymis stained with H&E (C, D) and testes stained with H&E (E, F), TUNEL (G, H) and anti-SCP3 antibody with DAPI (I, J). Data are representative of 4 independent experiments. Scale bars, 50 μm.
Fig 4.
Loss of PKnox1 causes accumulation of GFRα1+ cells and differentiation arrest of c-Kit+ spermatogonia.
Whole-mount immunodetection of cells expressing GFRα1 (red) and c-Kit (green) in seminiferous tubules of 12-week-old littermate controls (A, C, C’, E, G and G’), PKnox1-GKO (B, D), and -CKO mice (F, H). As; single cell, Apr; paired cells, Aal; aligned cells. Data are representative of 3 independent experiments. Scale bars, 50μm.
Fig 5.
PLZF and c-Kit are expressed in a distinct subset of spermatogonia in PKnox1-deficient testes.
Immunohistochemical analysis of PLZF (A, D) and c-Kit (B, E) expression in the testis from 12-week-old PKnox1-GKO and control mice with merged images (C, F). Tissue sections were double stained with the indicated combinations of antibodies and counterstained with DAPI. Arrowheads and arrows indicate PLZF+ and c-Kit+ cells, respectively. Broken lines indicate the basement membrane of seminiferous tubules. Data are representative of 3 independent experiments. Scale bars, 50 μm.
Fig 6.
Loss of PKnox1 affects PCNA expression in c-Kit+ spermatogonia.
Tissue sections from the testis from 12-week-old PKnox1-GKO and control mice were double stained with the indicated antibody combinations and counterstained with DAPI. Arrows indicate GFRα1+ and c-Kit+ cells with or without PCNA expression. Broken lines indicate the basement membrane of seminiferous tubules. Data are representative of 3 independent experiments. Scale bars, 25 μm.