ABSTRACT
Skeletal myogenesis is precisely regulated by multiple transcription factors. Previously, we demonstrated that enhancer of polycomb 1 (Epc1) induces skeletal muscle differentiation by potentiating serum response factor (SRF)-dependent muscle gene activation. Here, we report that an interacting partner of Epc1, ret finger protein (RFP), blocks skeletal muscle differentiation. Our findings show that RFP was highly expressed in skeletal muscles and was downregulated during myoblast differentiation. Forced expression of RFP delayed myoblast differentiation, whereas knockdown enhanced it. Epc1-induced enhancements of SRF-dependent multinucleation, transactivation of the skeletal α-actin promoter, binding of SRF to the serum response element, and muscle-specific gene induction were blocked by RFP. RFP interfered with the physical interaction between Epc1 and SRF. Muscles from rfp knockout mice (Rfp(-/-)) mice were bigger than those from wild-type mice, and the expression of SRF-dependent muscle-specific genes was upregulated. Myotube formation and myoblast differentiation were enhanced in Rfp(-/-) mice. Taken together, our findings highlight RFP as a novel regulator of muscle differentiation that acts by modulating the expression of SRF-dependent skeletal muscle-specific genes.
Subject(s)
DNA-Binding Proteins/metabolism , Muscle Cells/metabolism , Muscle Development/genetics , Nuclear Proteins/metabolism , Repressor Proteins/metabolism , Serum Response Factor/metabolism , Actins/genetics , Actins/metabolism , Animals , Binding Sites , Cell Differentiation , Cell Line , DNA-Binding Proteins/genetics , Gene Expression Regulation, Developmental , Mice , Mice, Knockout , Muscle Cells/cytology , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/metabolism , Myoblasts/cytology , Myoblasts/metabolism , Nuclear Proteins/genetics , Promoter Regions, Genetic , Repressor Proteins/genetics , Serum Response Factor/genetics , Transcription Factors , Transcriptional Activation , Ubiquitin-Protein LigasesABSTRACT
OBJECTIVE: To determine the incidence and characteristics of hyperventilation-induced nystagmus (HIN) in cerebellopontine angle (CPA) tumors and unilateral peripheral vestibulopathy (UPV), and to elucidate differential contribution of hyperventilation to bring out vestibular asymmetry between acute and chronic phases of UPV. METHODS: We recorded horizontal HIN in 33 patients with CPA tumors and 145 with UPV. The UPV included patients of either acute (7 days or less from symptom onset, n = 47) or chronic (more than 7 days from symptom onset, n = 98) phases. RESULTS: The incidence of HIN was higher in the CPA tumor than in the UPV group (82 vs 34%, p < 0.01) and was also higher in the acute than in the chronic UPV group (60 vs 21%, p < 0.01). Furthermore, HIN was more commonly ipsilesional (i-HIN) in the CPA tumor than in the UPV group (52 vs 8%, p < 0.01) and more commonly ipsilesional in the acute than in the chronic UPV group (21 vs 1%, p < 0.01). The patients with i-HIN and acoustic neuroma had a tendency to harbor smaller tumors and to have less severe caloric asymmetry. CONCLUSIONS: The contribution of hyperventilation on vestibular nystagmus differs depending on the disease phase or underlying pathologies. Our study demonstrates that hyperventilation-induced nystagmus (HIN) beating to the side of reduced caloric response, hearing impairment, or abnormal auditory brainstem response responses may be a valuable sign for bedside detection of cerebellopontine angle (CPA) tumors. CPA tumor should be a prime suspicion in patients with acute vertigo and ipsilesional HIN, especially when the vertigo accompanies hearing impairments.
Subject(s)
Cerebellar Neoplasms/physiopathology , Cerebellopontine Angle/physiopathology , Hyperventilation/physiopathology , Nystagmus, Pathologic/physiopathology , Vestibular Neuronitis/physiopathology , Adult , Aged , Cerebellar Neoplasms/diagnosis , Cerebellopontine Angle/pathology , Evoked Potentials, Auditory, Brain Stem/physiology , Female , Humans , Hyperventilation/complications , Male , Middle Aged , Nystagmus, Pathologic/etiology , Vestibular Neuronitis/diagnosisABSTRACT
Previously, PAHX-AP1 (PAHX-associated protein 1) was isolated as a novel protein to interact with Refsum disease gene product (phytanoyl-CoA alpha-hydroxylase, PAHX) and specifically expressed in mouse brain. PAHX-AP1 is also suggested to be involved in the development of the central neurologic deficits of Refsum disease. To clarify its function, we have searched for proteins that associate with PAHX-AP1 via yeast two-hybrid system. We found that PAHX-AP1 interacts with the cytoplasmic region of human brain-specific angiogenesis inhibitor 1 (hBAI1), and isolated murine homolog of hBAI1. Structural analysis of the PAHX-AP1 with three reported hBAI-associated proteins (BAP) revealed no homology among them, and we designated PAHX-AP1 as BAP4. The ability of BAP4 to interact with BAI1 was confirmed by pulling-down BAI1 with GST-BAP4 protein and immunoprecipitation study using brain lysate. Northern and Western blot analyses demonstrated a unique pattern of BAI1 expression in the brain. The peak level of BAI1 was observed 10 days after birth. In situ hybridization analyses of the brain showed the same localization of BAI1 as BAP4, such as most neurons of cerebral cortex, hippocampus, and V, VI, VII, VIII, and XII nuclei. Because BAI1 possessed thrombospondin-type 1 repeats in its extracellular region, changes of BAI1 expression were examined in the focal cerebral ischemia model. The BAI1 expression decreased on the ischemic side after 24 h but BAP4 was not changed after the time-course of ischemia. Our results indicate that expression and localization of BAI1 in the brain is correlated with BAP4, and that BAI1 is involved in inhibition of angiogenesis and neuronal differentiation.
