Fig. 4.

Aspm mutations cause major defects in the male and female germlines. (A) Testes from newborn (P0.5), juvenile (P21), and adult (10 wk) WT and 1-7 hom mice. (Scale bar, 2 mm.) (B) Boxplot of testis weight (sum of left and right testis) of adult (8–12 wk) WT, 1-25 and 1-7 het and hom, 1-7 hom +Hs, and +Hs mice. Data from (left to right) 34, 13, 13, 12, 16, 7, and 10 mice. (C and D) Double immunofluorescence for Aspm (green) with acetylated α-tubulin (acTub, red) in C, and with Aurora B (red) in D, combined with DAPI staining (blue), of 10-μm cryosections (epifluorescence) of an adult testis from a WT mouse. A spermatogenic cell in metaphase is shown in C, and in telophase in D. (Scale bar, 5 μm.) (E) Immunofluorescence for MVH (red) with DAPI staining (blue) of 10-μm cryosections (epifluorescence) of testes from P0.5 WT (Left) and 1-7 hom (Right) mice. In WT, numerous MVH+ cells are found in the seminiferous tubules, whereas in 1-7 hom, most tubules lack germ cells (white arrowheads), and only a few tubules contain MVH+ gonocytes (red arrowhead). (Scale bar, 50 μm.) (F and G) Quantification of seminiferous tubules containing MVH+ cells, expressed as a percentage of the total number of tubules contained in testis sections (F), and the average number of MVH+ cells per seminiferous tubule in testis sections (G), from P0.5 WT and 1-7 hom mice. Data are the mean of testes from three mice (sum of five sections per testis). (H) Immunofluorescence for MVH (red) combined with DAPI staining (blue) of 10-μm cryosections (epifluorescence) of testes, cut orthogonally to the longitudinal axis, from 10-wk-old WT (Left) and 1-7 hom (Right) mice. In WT, essentially every seminiferous tubule contains MVH+ spermatogenic cells, whereas in 1-7 hom, only approximately half of the tubules contain MVH+ cells. (Scale bar, 1 mm.) (I) DAPI staining of a 10-μm cryosection (epifluoresence) of a testis from a 10-wk-old 1-7 hom mouse, showing a tubule containing spermatogenic cells (single asterisk), and empty tubules (double asterisk) containing only few Sertoli cells, identified by the characteristic centromeric heterochromatin condensed in two chromocenters revealed by DAPI staining (Inset). (Scale bar, 100 μm, 10 μm in Inset.) (J) Immunofluorescence for Nobox (red) combined with DAPI staining (blue) of 10-μm cryosections (epifluorescence) of ovaries from adult WT (Left) and 1-7 hom (Right) mice. All follicular stages—primordial (PF), primary (1F), secondary (2F), and antral follicles—were observed in WT and 1-7 hom ovaries. (Scale bar, 50 μm.) (K) Boxplot of ovary weight (sum of left and right ovary) of adult (10–12 wk) WT and 1-25 and 1-7 hom mice. Data from (left to right) 7, 4, and 9 mice. (L) Quantification of Nobox+ cells per total ovary section area from adult (10–12 wk) WT and 1-7 hom mice. Data are the mean of ovaries from three mice (sum of five sections per ovary). In boxplots (B and K), the line within the box indicates the median value, the box spans the interquartile range, and whiskers extend to data extremes. **P < 0.01; ***P < 0.001.