Calcium-binding tyrosine phosphorylation-regulated protein （CABYR） is a highly polymorphic calcium-binding tyrosine- and serine-/threonine-phosphorylated fibrous sheath （FS） protein involved in capacitation. A put...Calcium-binding tyrosine phosphorylation-regulated protein （CABYR） is a highly polymorphic calcium-binding tyrosine- and serine-/threonine-phosphorylated fibrous sheath （FS） protein involved in capacitation. A putative domain （amino acids 12-48） homologous to the regulatory subunit of type II cAMP-dependent protein kinase A （RII） dimerisation and A kinase-anchoring protein （AKAP）-binding domains of protein kinase A at the N-terminus suggests that CABYR may self-assemble and bind to AKAPs. Moreover, there is evidence that CABYR has limited interaction with AKAPs. However, further evidence and new relationships between CABYR and other FS proteins, including AKAPs, will be helpful in understanding the basic physiology of FS. In this study, a new strategy for co-immunoprecipitation of insoluble proteins, as well as the standard co-immunoprecipitation method in combination with mass spectrometry and western blot, was employed to explore the relationship between CABYR, AKAP3 and Ropperin. The results showed that AKAP3 was co.immunoprecipitated with CABYR by the anti-CABYR-A polyclonal antibody, and, conversely, CABYR was also co.immunoprecipitated with AKAP3 by the anti-AKAP3 polyclonal antibody. Another RIl-like domain containing protein, Ropporin, was also co-immunoprecipitated with CABYR, indicating that Ropporin is one of CABYR＇s binding partners. The interactions between CABYR, AKAP3 and Ropporin were confirmed by yeast two-hybrid assays. Further analysis showed that CABYR not only binds to AKAP3 by its RII domain but binds to Ropporin through other regions besides the RIl-like domain. This is the first demonstration that CABYR variants form a complex not only with the scaffolding protein AKAP3 but also with another Rll-like domain-containing protein in the human sperm FS.展开更多
[ Objective] To study the effects of clindamycin on germ cells of male mice. [Method] A total of 48 healthy adult male mice were randomly divided into group A, group B, group C and control group, 12 mice in each group...[ Objective] To study the effects of clindamycin on germ cells of male mice. [Method] A total of 48 healthy adult male mice were randomly divided into group A, group B, group C and control group, 12 mice in each group. The mice in the group A, group B and group C were respectively administrated with clindamycin at doses of 15, 30 and 60 mg/kg.d via intraperitoneai injection, and those in the control group were trea- ted with normal saline. On Day 29 after administration, the mice were killed by dislocation, and the sperm motility rate, sperm deformity rate and the percentage of sperm having swollen tail membrane were determined, respectively. [ Result] The sperm motility rate and the percentage of sperm having swollen tail membrane of the group B and group C were lower than those of the control group; and the sperm deformity rate of the group B and group C was higher than that of the control group. The sperm motility rate, sperm deformity rate and percentage of sperm having swollen tail membrane had no significant difference between the group A and control group. [ Conclusion] The clindamycin at a dose equal to or more than that used for severe infection affects the normal function of sperm in mice.展开更多
文摘Calcium-binding tyrosine phosphorylation-regulated protein （CABYR） is a highly polymorphic calcium-binding tyrosine- and serine-/threonine-phosphorylated fibrous sheath （FS） protein involved in capacitation. A putative domain （amino acids 12-48） homologous to the regulatory subunit of type II cAMP-dependent protein kinase A （RII） dimerisation and A kinase-anchoring protein （AKAP）-binding domains of protein kinase A at the N-terminus suggests that CABYR may self-assemble and bind to AKAPs. Moreover, there is evidence that CABYR has limited interaction with AKAPs. However, further evidence and new relationships between CABYR and other FS proteins, including AKAPs, will be helpful in understanding the basic physiology of FS. In this study, a new strategy for co-immunoprecipitation of insoluble proteins, as well as the standard co-immunoprecipitation method in combination with mass spectrometry and western blot, was employed to explore the relationship between CABYR, AKAP3 and Ropperin. The results showed that AKAP3 was co.immunoprecipitated with CABYR by the anti-CABYR-A polyclonal antibody, and, conversely, CABYR was also co.immunoprecipitated with AKAP3 by the anti-AKAP3 polyclonal antibody. Another RIl-like domain containing protein, Ropporin, was also co-immunoprecipitated with CABYR, indicating that Ropporin is one of CABYR＇s binding partners. The interactions between CABYR, AKAP3 and Ropporin were confirmed by yeast two-hybrid assays. Further analysis showed that CABYR not only binds to AKAP3 by its RII domain but binds to Ropporin through other regions besides the RIl-like domain. This is the first demonstration that CABYR variants form a complex not only with the scaffolding protein AKAP3 but also with another Rll-like domain-containing protein in the human sperm FS.
基金supported by the grants from the University Scientific Research Project of Inner Mongolia Autonomous Region (NJzc08159)
文摘[ Objective] To study the effects of clindamycin on germ cells of male mice. [Method] A total of 48 healthy adult male mice were randomly divided into group A, group B, group C and control group, 12 mice in each group. The mice in the group A, group B and group C were respectively administrated with clindamycin at doses of 15, 30 and 60 mg/kg.d via intraperitoneai injection, and those in the control group were trea- ted with normal saline. On Day 29 after administration, the mice were killed by dislocation, and the sperm motility rate, sperm deformity rate and the percentage of sperm having swollen tail membrane were determined, respectively. [ Result] The sperm motility rate and the percentage of sperm having swollen tail membrane of the group B and group C were lower than those of the control group; and the sperm deformity rate of the group B and group C was higher than that of the control group. The sperm motility rate, sperm deformity rate and percentage of sperm having swollen tail membrane had no significant difference between the group A and control group. [ Conclusion] The clindamycin at a dose equal to or more than that used for severe infection affects the normal function of sperm in mice.
