Menkes disease is an effect of ATP7A gene mutation in humans, coding the Cu-ATP-ase which is essential in intestinal copper absorption and its subsequent transfer to circulation. This mutation results in a deficiency of copper in all tissues except the epithelia of intestine and kidney tubules. Subcutaneous injection of copper ions is the main therapy for Menkes patients. Mosaic (Atp7a(mo-ms)) mice closely simulate the situation in Menkes disease. The aim of this study was to evaluate the changes in structure and element content in kidneys of mosaic mice after copper supplementation. Hematoxylin-eosin staining was used to analyze tissue morphology and atomic absorption spectrometry to estimate Cu and Zn content. X-ray microanalysis was performed to measure Na, Mg, P, Cl, and K content in the cells of the proximal and distal tubules. Copper administration lengthened the lifespan of the mutants but led to its high accumulation and results in severe kidney damage. Karyomegalia, necrosis of tubular and Bowman's capsule epithelium, lesions, and atrophy of glomeruli were observed in the treated mutants. Copper treatment afterwards led to sclerosis of glomeruli and tubules enhanced proliferation of epithelial cells and formation of both polycystic and papillary carcinoma patterns in kidney. We suggest that copper excess may impair the activity of Na(+)/K(+) ATP-ase in renal tubules of ms/- males. The content of Mg, P, and Cl in kidneys in mutants was also changed after copper administration.

Two congenic strains of mice (control, B10.BR/SgSn; mutant, B10.BR-Ydel/Ms with partial deletion of the Y chromosome) were examined. In control males, 22.6% of spermatozoa had abnormal heads; in mutant males, there were 64.2%, the most common being heads with flat acrosomes. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of mature sperm proteins, followed by acrosin assay and acrosome silver staining, revealed a reduced concentration of acrosin in acrosomal caps in 35.8% of the spermatozoa in mutant males. Electron microscope analysis showed that some of the round, early spermatids in the mutants had normally formed acrosomal caps but lacked the proacrosomal granule and had no, or only scarce, acrosomal material. These observations indicate that formation of the acrosomal cap is controlled separately from the synthesis of the acrosomal material and suggest that some factors linked on the Y chromosome are involved in the control of acrosome development.

Two congenic lines of mice, one with a partial deletion of the Y chromosome, differ in the percentage of spermatozoa with abnormal heads: B10.BR/SgSn males give 22-6% and B10.BR-Y del /Ms males give 64-2% abnormal sperm. The FjS resulting from crosses of B10.BR/SgSn males with females of five common inbred strains exhibited significantly lower levels of abnormal sperm than the parental strains, as opposed to F x hybrids sired by B10.BR-Y deI /Ms mutant males, where very high levels of abnormal spermatozoa were found. About 30% of abnormal spermatozoa, produced by males with deletion on the Y chromosome, were characterized by a flat acrosomal cap. This class of abnormality was never observed in non-mutant males, suggesting a mutant-specific defect. These results demonstrate the important role of the Y chromosome in spermatogenesis.

Males of the mouse strain B10.BR/SgSn and its congenic mutant strain B10.BR-Ydel, with a partial deletion of the Y chromosome, were used to examine factors related to poor sperm quality and quantity in the mutant strain. The testes of males from the two strains did not differ in their immunohistochemical reaction to androgen receptors or in the number of Sertoli and germ cells in tubules with normal morphology. However, mutants showed a greater frequency of degenerated tubules, a higher level of X-Y chromosome dissociation at meiosis (18% v. 10% in control males), and a lower content of resistant sperm heads in testis homogenates. In the cauda epididymidis, there was a higher percentage of spermatozoa with abnormal heads (88% v. 31%) and of spermatozoa with a cytoplasmic droplet still attached (74% v. 51%). Many sperm heads with flat acrosomes, occurring only in mutants (30% of sperm population), were deficient in proteolytic enzymes, as evidenced by the reaction on gelatine membranes. Most copulations of mutant males (11/18) were sterile in spite of the presence of spermatozoa in the uterus, but in the remaining copulations the fertilization rate was reasonably good (79%). Low numbers of spermatozoa were recovered from the oviducts, and those with the most severely deformed heads were less frequent there than in the uterus. The results show that a partial deletion of the Y chromosome affects efficiency of spermatogenesis, morphology of spermatozoa, their epididymal maturation and capacity to reach the ampulla and fertilize eggs.

In the present study, animals with a genetic defect in copper metabolism were used as a model organism to study the role of copper in reproduction and to determine whether the disturbances in copper and zinc metabolism affect the testicular tissue and gamete quality in males. Mice with an X-linked mosaic mutation (Atp7a(mo-ms)) exhibit pathological features characteristic of affected copper metabolism. This mutation usually leads to lethality of the mutant males which generally expire on about day 16. Only 4% of mutant animals survive the critical period, achieve maturity, and become fertile. To improve the mutants' viability they were treated with subcutaneous injections of cupric chloride. We measured copper and zinc concentration in the gonads of young (14-day-old) and adult (5-month-old) mutant and control males. Results indicate that copper content was increased but zinc was decreased in the mutant testes. Analysis of the morphology of the testis of the young animals indicate that apoptosis (characteristic for the gonads of young males) was increased in the gonads of the 14-day-old mutants. This process was less advanced in the group of 14-day-old copper treated control males. Apoptosis was also increased in the testes of the adult mutants. Moreover in adult mutants we observed pathological changes in testes morphology (atrophic and sclerotic tubules). Copper and zinc disorders also negatively influenced semen quality parameters, including sperm motility, head morphology, tail cytoplasmic membrane integrity, and number of viable spermatozoa. Poor semen quality of the mutant males seems to be responsible for affected in vivo fertilization efficiency. Treatment with cupric chloride did not influence semen quality except in maturation rate, which was even slower in both mutant and control males after treatment. Additionally, in mutants, copulatory plugs and fertile copulation outcome were decreased after copper treatment.