RESUMEN
In our previous study (Kawashima et al., Biol Reprod 2009; 80: 1293-1304), we suggested that the first cycle of spermatogenesis recovered from busulfan-induced temporary arrest was a good model system to analyze the proteins expressed at the specific stages of spermatogenesis in the mouse, and this has been confirmed in the present paper. Namely, six-week-old mice were injected with busulfan at 20 mg/kg body weight. The germ cells except for the undifferentiated spermatogonia disappeared by 32 days after injection. The surviving spermatogonia started to proliferate, and spermatogenesis was entirely recovered about 77 days after injection. By proteome analysis of the busulfan-treated testis during the process of recovery of spermatogenesis, we identified a protein that was expected to be expressed in the spermatogenic cells as Superkiller viralicidic activity-2-like-2 (SKIV2L2). Skiv2l2 mRNA was found in the kidney, epididymis and heart as well as the testis. In the testis, Skiv2l2 mRNA was shown to be highly expressed in the spermatocytes at stages I to VI. On the other hand, SKIV2L2 protein was found to be predominantly localized in the nuclei of round spermatids. In accordance with the fact that SKIV2L2 belongs to the Ski2 family within the superfamily 2 of RNA helicases, it has been shown that SKIV2L2 has both the RNA-binding and ATPase activities.
Asunto(s)
Núcleo Celular/enzimología , Proteínas Nucleares/biosíntesis , ARN Helicasas/metabolismo , Proteínas de Unión al ARN/biosíntesis , Espermátides/enzimología , Animales , Peso Corporal , Epidídimo/enzimología , Hibridación in Situ , Riñón/enzimología , Masculino , Ratones , Miocardio/enzimología , Proteínas Nucleares/genética , Proteoma , Proteómica/métodos , Proteínas de Unión al ARN/genética , Espermatogonias/enzimología , Testículo/enzimologíaRESUMEN
The occurrence of dreaming during rapid eye movement (REM) sleep prompts interest in the role of REM sleep in hippocampal-dependent episodic memory. Within the mammalian hippocampus, the dentate gyrus (DG) has the unique characteristic of exhibiting neurogenesis persisting into adulthood. Despite their small numbers and sparse activity, adult-born neurons (ABNs) in the DG play critical roles in memory; however, their memory function during sleep is unknown. Here, we investigate whether young ABN activity contributes to memory consolidation during sleep using Ca2+ imaging in freely moving mice. We found that contextual fear learning recruits a population of young ABNs that are reactivated during subsequent REM sleep against a backdrop of overall reduced ABN activity. Optogenetic silencing of this sparse ABN activity during REM sleep alters the structural remodeling of spines on ABN dendrites and impairs memory consolidation. These findings provide a causal link between ABN activity during REM sleep and memory consolidation.
Asunto(s)
Condicionamiento Psicológico , Giro Dentado/fisiología , Consolidación de la Memoria/fisiología , Neuronas/fisiología , Sueño REM/fisiología , Animales , Calcio/metabolismo , Giro Dentado/citología , Electroencefalografía , Electromiografía , Miedo , Hipocampo , Aprendizaje , Ratones , Neurogénesis , Optogenética , Ritmo TetaRESUMEN
Single intraperitoneal injection of busulfan at 20 mg/kg body weight to mature male mice induced the deletion of the spermatogenic cells, followed by the restoration of the spermatogenesis by the surviving undifferentiated spermatogonia. The changes of the protein contents in testis during these processes were analyzed by two-dimensional gel electrophoresis in order to identify the proteins expressed at the specific stages of spermatogenesis. An acidic protein that disappeared and recovered in the same time course as spermatids after the busulfan treatment was identified as CABS1 by mass spectrometry. It was found that CABS1 was specifically expressed in the elongate spermatids at steps 13 to 16 in stages I to VIII of the seminiferous epithelium cycle of the mouse, and then it localized to the principal piece of flagellum of the mature sperm in the cauda epididymis. We have found for the first time that CABS1 is a calcium-binding protein that binds calcium during the maturation in the epididymis.