AIMTo characterize mouse capping protein alpha3 (CPalpha3) during spermatogenesis and sperm maturation.

METHODSWe produced rat anti-CPalpha3 antiserum and examined the expression of CPalpha3 in various mouse tissues using Western blot analysis and the localization of CPalpha3 in testicular and epididymal sperm using immunohistochemical analyses. We also examined how the localization of CPalpha3 and beta-actin (ACTB) in sperm changed after the acrosomal reaction by performing immunohistochemical analyses using anti-CPalpha3 antiserum and anti-actin antibody.

RESULTSWestern blot analysis using specific antiserum revealed that CPalpha3 was expressed specifically in testes. Interestingly, the molecular weight of CPalpha3 changed during sperm maturation in the epididymis. Furthermore, the subcellular localization of CPalpha3 in sperm changed dynamically from the flagellum to the post-acrosomal region of the head during epididymal maturation. The distribution of ACTB was in the post-acrosomal region of the head and the flagellum. After inducing the acrosomal reaction, the CPalpha3 and ACTB localization was virtually identical to the localization before the acrosomal reaction.

CONCLUSIONCPalpha3 might play an important role in sperm morphogenesis and/or sperm function.

Acrosome Reaction ; physiology ; Actins ; metabolism ; Animals ; Blotting, Western ; CapZ Actin Capping Protein ; metabolism ; Cells, Cultured ; Epididymis ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Sperm Head ; metabolism ; Sperm Tail ; metabolism ; Spermatogenesis ; physiology ; Spermatozoa ; cytology ; metabolism ; Testis ; cytology ; metabolism

AIMTo investigate the associations of autosomal and X-chromosome homologs of the RNA-binding-motif (RNA-binding-motif on the Y chromosome, RBMY) gene with non-obstructive azoospermia (NOA), as genetic factors for NOA may map to chromosomes other than the Y chromosome.

METHODSGenomic DNA was extracted using a salting-out procedure after treatment of peripheral blood leukocytes with proteinase K from Japanese patients with NOA (n=67) and normal fertile volunteers (n=105). The DNA were analyzed for RBMX by expressed sequence tag (EST) deletion and for the like sequence on chromosome 9 (RBMXL9) by microsatellite polymorphism.

RESULTSWe examined six ESTs in and around RBMX and found a deletion of SHGC31764 in one patient with NOA and a deletion of DXS7491 in one other patient with NOA. No deletions were detected in control subjects. The association study with nine microsatellite markers near RBMXL9 revealed that D9S319 was less prevalent in patients than in control subjects, whereas D9S1853 was detected more frequently in patients than that in control subjects.

CONCLUSIONWe provide evidence that deletions in or around RBMX may be involved in NOA. In addition, analyses of markers in the vicinity of RBMXL9 on chromosome 9 suggest the possibility that variants of this gene may be associated with NOA. Although further studies are necessary, this is the first report of the association between RBMX and RBMXL9 with NOA.

Adult ; Chromosomes, Human, Pair 9 ; genetics ; Chromosomes, Human, X ; genetics ; Expressed Sequence Tags ; Heterogeneous-Nuclear Ribonucleoproteins ; genetics ; Humans ; Male ; Microsatellite Repeats ; genetics ; Nuclear Proteins ; genetics ; Oligospermia ; genetics ; Polymorphism, Genetic ; RNA-Binding Proteins ; genetics