Types A and B Niemann-Pick disease (NPD) are lysosomal storage disorders resulting from the deficient activity of acid sphingomyelinase (ASM). In this manuscript we report the pathobiology of male gonadal tissue and sperm in a knockout mouse model of NPD and demonstrate the importance of ASM for normal sperm maturation and function. Characteristic lipid-filled vacuoles were evident in light micrographs of testis' seminiferous tubules and epithelial cells lining the epididymis of ؊/؊ mice. Electron micrographs extended these findings and revealed storage vesicles within Sertoli cells of the seminiferous tubules. Mature spermatozoa from ؊/؊ mice showed marked ASM deficiency and elevated levels of sphingomyelin and cholesterol. Flow cytometric analysis revealed that affected spermatozoa had disrupted plasma and acrosome membranes, and mitochondrial membrane depolarization. They also did not undergo proper capacitation. Morphological abnormalities such as kinks and bends at the midpieceprinciple piece junction were evident in spermatozoa from affected mice, with consequent deficits in motility. Notably, the mutant sperm regained normal morphology on incubation in mild detergent, demonstrating that the bending defects were a direct consequence of membrane lipid accumulation. A mechanism for these abnormalities is proposed that suggests lipid accumulation in the gonads results in regulatory volume decrease defects within the developing sperm, and that regulatory volume decrease defects, in turn, lead to the observed abnormalities in sperm morphology and function. These results provide in vivo evidence that ASM activity plays a critical role in sperm maturation and function, and a basis for similar studies in sexually mature, male NPD patients. 1 Type A NPD is a severe, neurodegenerative form of the disorder that generally leads to death by ϳ3 years of age. In contrast, patients with type B NPD have little or no neurological involvement and may survive into adolescence or adulthood. ASM belongs to a family of sphingomyelinases that catabolize sphingomyelin (SPM) to ceramide and phosphorylcholine. 2,3 It is presumed that the pathophysiology in NPD is primarily because of the accumulation of SPM and other metabolically related lipids (eg, cholesterol) within the cells and tissues of affected patients, although signaling through the ceramide pathway may also be disrupted.

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Acid sphingomyelinase knockout mice are a model of the inherited human disorder types A and B NiemannPick disease. Herein, we show that heterozygous (ASMKO ؉/؊ ) mice have two distinct sperm populations resembling those found in normal and mutant animals, respectively, and that these two populations could be distinguished by their morphology, ability to undergo capacitation or the acrosome reaction, and/or mitochondrial membrane potential (MMP). The abnormal morphology of the mutant sperm could be normalized by demembranation with detergents or by the addition of recombinant acid sphingomyelinase to the culture media, and the corrected sperm also had an enhanced fertilization capacity. Methods were then explored to enrich for normal sperm from the mixed ASMKO ؉/؊ population, and flow cytometric sorting based on MMP provided the best results. In vitro fertilization was performed using ASMKO ؉/؊ oocytes and sperm before and after MMP sorting, and it was found that the sorted sperm produced significantly more wild-type pups than nonsorted sperm. Sperm sorting is much less invasive and more cost-effective than egg isolation, and offers several advantages over the existing assisted reproduction options for Niemann-Pick disease carrier couples. It therefore could have a major impact on the prevention of this and perhaps other genetic diseases. (Am J

PROBLEM: The purpose of this study was to investigate the levels of interleukin‐6 and −8 (IL‐6 and IL‐8) in sera and cervical mucus of infertile (idiopathic and immunoinfertile) women and to compare the levels with those in age‐matched normally cycling fertile women.METHODS: Levels of IL‐6 and IL‐8 were measured in the sera and cervical mucus of fertile and infertile women by the enzyme‐linked immunosorbent assay (ELISA). A non‐parametric Mann‐Whitney test was used to evaluate significance between the means. Linear regression analysis of IL‐6 and IL‐8 concentrations in serum versus cervical mucus and with antisperm antibody titers was performed by condition as well as all groups together. P < 0.05 was considered significant.RESULTS: Both IL‐6 and IL‐8 were detected in sera and cervical mucus of fertile as well as infertile groups. Although serum levels of IL‐6 and IL‐8 were significantly different between fertile and infertile groups, the differences were more pronounced in the cervical mucus samples. Cervical mucus of idiopathic and immunoinfertile women demonstrated significantly (P = 0.002 to < 0.0001) greater concentrations of IL‐6 and IL‐8 compared to fertile controls. In general, there was no significant correlation between the serum and cervical mucus IL‐6 and IL‐8 levels, whether analyzed by condition or all groups together.CONCLUSIONS: Elevated levels of IL‐6 and IL‐8 in cervical mucus of infertile groups may play a role in etiology of idiopathic and immunologic infertility. These findings suggest that the measurements of cytokines (especially IL‐6 and IL‐8) in cervical mucus may provide a tool for specific diagnosis and possibly treatment of infertility, especially idiopathic infertility.

Genetic diagnosis of preimplantation embryos (PGD) can substantially reduce the chance that at-risk couples have children afflicted with inherited diseases. However, PGD requires DNA,which is usually obtained from single cells following embryo biopsy. In addition, PGD requires that the genetic defect(s) causing the disorder be known. We have therefore developed an alternative to PGD, which we term preimplantation enzymatic diagnosis (PED). PED has several advantages over PGD, including the facts that it does not require embryo biopsy and that the gene defect(s) causing the disorder need not be known. We have demonstrated 'proof of principle' for this approach using embryos obtained from a mouse model (ASMKO mice) of acid sphingomyelinase (ASM)-deficient Niemann-Pick disease, an inherited lysosomal storage disorder. For this technique, fluorescently (BODIPY)-conjugated sphingomyelin was used to detect ASM activity in situ. Wild-type, preimplantation embryos degraded the substrate following a short 'pulse-chase' period, resulting in markedly reduced fluorescence compared to ASMKO embryos, which retained the fluorescent substrate. Thus, the two embryo types could be easily distinguished by fluorescent microscopy. The fluorescent sphingomyelin was not toxic to the embryos, and the entire procedure could be accomplished within 48 h without embryo biopsy. We suggest that PED may be useful for the preimplantation diagnosis of lysosomal storage disorders, and perhaps other enzymatic defects where similar in situ assay methods are available.