Targeted disruption of the mouse ADAMTS-1 gene. (a) Genomic locus, targeting vector, and predicted targeted locus. Black boxes represent exons 1–9. The probe for Southern analysis is indicated. B, BamHI; E, EcoRI; G, BglII. (b and c) Southern analysis of genomic DNA from targeted ES cells (b) and the offspring from a heterozygote intercross (c). The 7.0-kb EcoRI fragment and the 11.5-kb BamHI fragment denote the homologous recombinant allele. (d) RT-PCR of total RNA extracted from the kidney of each genotype. No ADAMTS-1 expression is detected in ADAMTS-1^–/– mice.

Growth retardation in ADAMTS-1^–/– mice. (a) Appearance of 1-week-old ADAMTS-1^+/+ (left) and ADAMTS-1^–/– (right) littermates. (b) Comparison of growth after birth between ADAMTS-1^+/+ (n = 10) and ADAMTS-1^–/– (n = 9) littermates. ^AP < 0.05 vs. wild-type littermates. (c) Comparison of body weight among 9-month-old ADAMTS-1^+/+, ADAMTS-1^+/–, and ADAMTS-1^–/– littermates (n = 13 in each genotype). ^BP < 0.0001. (d and e) Epididymal fat pads and testes from 12-week-old ADAMTS-1^+/+ (d) and ADAMTS-1^–/– (e) littermates. B, bladder; E, epididymis; F, fat pad; T, testis.

Comparison of kidneys between ADAMTS-1^+/+ (a, c, e, g) and ADAMTS-1^–/– (b, d, f, h, i) mice at 4 weeks. (a and b) Inner aspects of the kidneys. Arrow, renal papilla; arrowhead, fibrous tissue surrounding the ureteropelvic junction. (c and d) Hematoxylin and eosin–stained sections of the whole kidney. CS, caliceal space. (e and f) Mallory-azan–stained sections through the renal hilum; the ureteropelvic junction of ADAMTS-1^–/– mice is rich in fibrous tissue (arrows). P, renal papilla. (g–i) Electron microscopy findings at the ureteropelvic junction in ADAMTS-1^–/– mice and controls. (h) Excessive collagen fibers (arrows) between smooth muscle cells (Sm) at the ureteropelvic junction of ADAMTS-1^–/– mice. (i) Thickening of the collagenous segment between the epithelial cell (Epi) layer and smooth muscle cell layer (12–30 μm), and proliferation of fibroblasts at the ureteropelvic junction of ADAMTS-1^–/– mice. (g) Collagenous segment between the epithelial cell layer and smooth muscle cell layer (6–12 μm) from the ureteropelvic junction of normal mice. (j–m) Electron microscopy ureteral findings in ADAMTS-1^–/– mice (k–m) and controls (j). (k) Thickening of the collagenous segment (arrows) between the epithelial cell layer and smooth muscle cell layer. (l) High magnification showing the accumulation of excessive collagen fibers in ADAMTS-1^–/– mice. (m) Deformity and fissures (arrowheads) in smooth muscle cells in ADAMTS-1^–/– mice. Bars in e and f represent 20 μm; bars in g–m represent 2 μm.

Intravenous pyelography (a–d). X-ray films were taken 5 minutes (a and c) and 30 minutes (b and d) after injection of contrast medium into 6-week-old ADAMTS-1^+/+ (a and b) and ADAMTS-1^–/– (c and d) littermates. In ADAMTS-1^+/+ mice, normal nephrograms were detected at 5 minutes; these disappeared within 30 minutes. In ADAMTS-1^–/– mice, the enlarged and distorted calyces still retained contrast medium after 30 minutes. Arrowheads and arrows indicate nephrograms and cystograms, respectively.

Immunohistochemistry of a mouse kidney section using an anti–ADAMTS-1 antibody (a) and a nonimmune control (b). J, urinary epithelium at the ureteropelvic junction. The pink staining shows the location of ADAMTS-1 expression in the urinary epithelium at the ureteropelvic junction.

Comparison of the adrenal glands between ADAMTS-1^+/+ (a and c) and ADAMTS-1^–/– (b and d) mice at 4 weeks of age. (a and b) Hematoxylin and eosin–stained sections of the adrenal glands. C, adrenal cortex; M, adrenal medulla. (c, d) High-magnification light micrographs of the adrenal medulla adjacent to the corticomedullary junction (J). Bars in a and b represent 20 μm; bars in c and d represent 200 μm.

Comparison of uterus and ovary between ADAMTS-1^+/+ (a, c, e) and ADAMTS-1^–/– (b, d, f) female mice at 12 weeks. (a and b) Exterior appearance of uterus and ovary. Hematoxylin and eosin–stained sections of uterus (c and d) and ovary (e and f). Arrows in d represent abnormal cystic formation in the uterus of ADAMTS-1^–/– mice. Bars in c–f represent 100 μm.