Expression and localization of the deubiquitinating enzyme mUBPy in wobbler mouse testis during spermiogenesis.
Mouse ubiquitin-specific processing protease (mUBPy) is a deubiquitinating enzyme highly expressed in both brain and testis. In testis, it interacts with the DnaJ protein, MSJ-1; both mUBPy and MSJ-1 are located on the cytoplasmic surface of the developing acrosome and in the centrosomal region during spemiogenesis. Present data show the first appearance in testis of mUbpy mRNA and protein at 10 days post-partum (d.p.p.). In addition, to investigate on a possible role of mUBPy in sperm formation, we took advantage of mutant wr/wr (wobbler) mice characterized by male infertility, which is likely due to the lack of a real, functional acrosome. RT-PCR and Northern blot analyses show that mUbpy is up-regulated in adult wobbler testis. Furthermore, in wild-type testis mUBPy protein is primarily detected by Western blot in the soluble (cytosolic/nuclear) fraction during the first round of spermatogenesis and in the adult. By contrast, mUBPy is primarily detected in membranous/insoluble protein fraction when wobbler phenotype is clearly shown (30 d.p.p.) and in adult wobbler testis. By immunohistochemistry, whereas in wild-type animals mUBPy marks the profile of the acrosomic vesicle in differentiating spermatids, in wobbler mice only a detergent pre-treatment procedure allows to detect mUBPy immunoreactivity, which results in diffuse spotted granules inside the cytoplasm and around the nuclear shape. In conclusion, in wobbler testis expression of mUbpy is up-regulated, while a differential sorting of the protein characterizes wobbler spermatids where acrosome formation is impaired.
Chianese
R
R
Dipartimento di Medicina Sperimentale, Seconda Università di Napoli, Italy.
Scarpa
D
D
Berruti
G
G
Cobellis
G
G
Pierantoni
R
R
Fasano
S
S
Meccariello
R
R
D000177:Acrosome / Q000201:enzymology / Q000502:physiology; D000818:Animals; D010450:Endopeptidases / Q000032:analysis* / Q000235:genetics* / Q000502:physiology; D056827:Endosomal Sorting Complexes Required for Transport / Q000032:analysis* / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008818:Mice, Neurologic Mutants; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis; D013087:Spermatids / Q000201:enzymology; D013091:Spermatogenesis / Q000502:physiology*; D013737:Testis / Q000201:enzymology* / Q000254:growth & development; D043222:Ubiquitin Thiolesterase / Q000032:analysis* / Q000235:genetics* / Q000502:physiology
D056827:Endosomal Sorting Complexes Required for Transport; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D010450:Endopeptidases; D043222:Ubiquitin Thiolesterase; C541536:Usp8 protein, mouse