Project description:Neurovascular dysfunction in penis is a fundamental reason of erectile dysfunction. Diabetes mellitus is one of the major causes of erectile dysfunction and leads to a poor response to oral phosphodiesterase-5 inhibitors. Heat shock protein 70 (Hsp70), a ubiquitous molecular chaperone exist in all living organisms, is known to play a role in cell survival and neuroprotection. Here, we report an effectiveness of Hsp70 in mediating neurovascular regeneration in diabetic conditions. Using Hsp70-Tg mice or Hsp70 protein administration, we demonstrate that overexpression of Hsp70 in diabetic mice restores erectile function through enhanced penile angiogenesis and neural regeneration. We found that cystathionine gamma-lyase (Cse) is a novel target of Hsp70-driven penile angiogenesis and neural regeneration. Hsp70-Cse triggered SDF1/HO-1/PI3K/Akt/eNOS/NF-κB p65 pathways involved in angiogenesis and neural regeneration. Coimmunoprecipitation and His-Tag pull down assay using mouse cavernous endothelial cells treated with Hsp70 showed the physical interaction between Hsp70 and Cse, and solid-phase binding assay revealed a high-affinity Hsp70-Cse binding with an apparent dissociation constant of 1.8 nmol/L. We provide a novel and solid evidence for Cse-dependent mechanism of Hsp70, which mediates Hsp70-induced neurovascular regeneration and the restoration of erectile function under diabetic conditions.
Project description:Round cell component is a resk factor associated with metastasis and poor prognosis in myxoid liposarcoma. To identify microRNAs which play a role in the malignancy of round cell component, we performed microRNA microarray analysis and compared the miRNA expression profilings between myxoid and round cell component.
Project description:Gene expression during spermatogenesis undergoes significant changes due to a demanding sequence of mitosis, meiosis and differentiation. Round spermatids have to prepare for DNA condensation as well as synthesize transcripts of genes required for the differentiation process. However, the mechanisms of gene activation and silencing in round spermatids remain largely unknown. We employed ChIP-seq to figure out the correlation of H3K4me3 and H3K9me3 marks with transcriptional activation and silencing in round spermatids. Out of about 2800 genes identified by H3K4me3 chipseq, transcriptome sequencing in purified round spermatids showed the presence of transcripts corresponding to about 64% of the genes. On the other hand, only about 25% of H3K9me3 enriched genes showed transcription in round spermatids, that too at very low levels. In conclusion, H3K4me3 enrichment in round spermatids correlates significantly with gene expression and H3K9me3 correlates with gene silencing, both of which are equally important for successful spermatogenesis.
Project description:We examined the RNA binding ability of the 70-kDa heat shock protein (HSP70) family member HSPA1A and identified the RNA species bound to HSP70 in vivo. We used a UV-crosslinkiung and immunoprecipitation based approach named FLASH-seq (Aktaş et al., 2017) using 3xFLAG-Histidine-Biotin tagged ectopically expressed protein in HEK293T cells.