Rat hypodactyly (hd) mutation is characterized by abnormal spermatogenesis and sperm decapitation, limb malformation (missing digits II and III) and growth retardation. We have previously reported centrobin (centrosome BRCA2-interacting protein) truncation at the C-terminus in the hd mutant. Here, we report data from a transgenic rescue experiment carried out to determine a role of centrobin in pathogenesis of hd. The transgenic construct, consisting of full-length-coding cDNA linked to a ubiquitous strong promoter/enhancer combination, was inserted to chromosome 16 into a LINE repeat. No known gene is present in the vicinity of the insertion site. Transgenic centrobin was expressed in all tissues tested, including testis. Transgenic animals show normal body weight and limb morphology as well as average weight of testis and epididymis. Yet, abnormal spermatogenesis and sperm decapitation persisted in the transgenic animals. Western blotting showed the coexistence of full-length and truncated or partially degraded centrobin in sperm of the rescued transgenic animals. Immunocytochemistry showed a buildup of centrobin and ODF2 (outer dense fiber 2) at the sperm decapitation site in the hd mutant and rescued transgenic rats. Additional findings included bulge-like formations and thread-like focal dissociations along the sperm flagellum and the organization of multiple whorls of truncated sperm flagella in the epididymal lumen. We conclude that centrobin is essential for normal patterning of the limb autopod. Centrobin may be required for stabilizing the attachment of the sperm head to flagellum and for maintaining the structural integrity of the sperm flagellum. We postulate that the presence of truncated centrobin, coexisting with full-length centrobin, together with incorrect timing of transgenic centrobin expression may hamper the rescue of fertility in hd male rats.

• MeSH
• Epididymis pathology   MeSH
• Gene Expression MeSH
• Fertility genetics   MeSH
• Homeodomain Proteins genetics   MeSH
• Rats MeSH
• Mutation * MeSH
• Mice MeSH
• Rats, Transgenic MeSH
• Cell Cycle Proteins genetics   metabolism   MeSH
• Heat-Shock Proteins metabolism   MeSH
• Spermatozoa growth & development   metabolism   MeSH
• Testis pathology   MeSH
• Protein Transport MeSH
• Organ Size genetics   MeSH
• Limb Deformities, Congenital genetics   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• Cntrob protein, rat MeSH Browser
• homeobox protein HOXA13 MeSH Browser
• Homeodomain Proteins MeSH
• Odf2 protein, rat MeSH Browser
• Cell Cycle Proteins MeSH
• Heat-Shock Proteins MeSH