Cell-Free Seminal mRNA and MicroRNA Exist in Different FormsReportar como inadecuado




Cell-Free Seminal mRNA and MicroRNA Exist in Different Forms - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Background

The great interest in cell-free mRNA, microRNA miRNA as molecular biomarkers for clinical applications, and as ‘signaling’ molecules for intercellular communication highlights the need to reveal their physical nature. Here this issue was explored in human cell-free seminal mRNA cfs-mRNA and miRNA cfs-miRNA.

Methodology-Principal Findings

Selected male reproductive organ-specific mRNAs, miRNAs, and piRNAs were quantified by quantitative real-time PCR in all experiments. While the stability of cfs-miRNA assessed by time-course analysis up to 24 h at room temperature was similar with cfs-mRNA, the reductive changes between cfs-miRNA and cfs-mRNA after filtration and Triton X-100 treatment on seminal plasma were very different, implying their different physical nature. Seminal microvesicles SMVs were then recovered and proportions of cfs-mRNA and cfs-miRNA within SMVs were quantified. The amounts of SMVs- sequestered cfs-mRNAs almost were the same as total cfs-mRNA, and were highly variable depending on the different sizes of SMVs. But most of cfs-miRNA was independent of SMVs and existed in the supernatant. The possible form of cfs-miRNA in the supernatant was further explored by filtration and protease K digestion. It passed through the 0.10-µm pore, but was degraded dramatically after intense protease K digestion.

Conclusions-Significance

The predominant cfs-mRNA is contained in SMVs, while most cfs-miRNA is bound with protein complexes. Our data explained the stability of extracellular RNAs in human semen, and shed light on their origins and potential functions in male reproduction, and strategy of developing them as biomarkers of male reproductive system.



Autor: Honggang Li, Shiyun Huang, Cuicui Guo, Huangtao Guan, Chengliang Xiong

Fuente: http://plos.srce.hr/



DESCARGAR PDF




Documentos relacionados