Project description:Total RNAs were extracted from the purified cauda epididymal spermatozoa of mouse (C57BL/6, 2 months old) and rats (Sprague Dawley, 450g young adult), and the 18 - 45 nt fraction small RNAs were subjected to library construction and deep sequencing, using Illumina GAIIx.
Project description:Total RNAs were extracted from the purified caput epididymal spermatozoa of mouse (C57BL/6, 2 months old) and rats (Sprague Dawley, 450g young adult), and the 18 - 45 nt fraction small RNAs were subjected to library construction and deep sequencing, using Illumina HiSeq 2000.
Project description:Total RNAs were extracted from the Testis and Epididymal Caput, Corpus and Cauda tissues of 2-month and 13-month-old WT and Gpx5 KO mice (C57BL/6). The 18 - 40 nt fraction small RNAs and transcriptomes were subjected to library construction and deep sequencing, using Illumina GAIIx or Hiseq 2000.
Project description:While assisted reproductive technologies (ARTs) are widely used in domestic animals, successful implementation of ARTs to conserve wildlife species remains challenging. In macropods, crucial aspects of fundamental reproductive biology, including changes induced by epididymal maturation, remain unknown, limiting the development of ARTs. In this context we performed a proteomic analysis of spermatozoa from the caput, corpus, and cauda epididymis of Eastern Grey Kangaroos (n = 6) to profile changes over epididymal maturation. Samples prepared by FASP digestion were analysed by LC-MS/MS with SWATH acquisition. A total of 4,304 proteins were identified, with significant overlap across epididymal regions. Highly abundant proteins in common across caput, corpus and cauda spermatozoa had strong enrichment for tubulins and included 4 histone proteins. The most significant proteomic remodelling was observed in the corpus to cauda transition, late in epididymal transit (728 differentially abundant proteins). Overall proteomic changes across epididymal maturation (1,131 differentially abundant proteins) suggested a loss of sperm glycosidases and an increase in flagellar proteins, including tubulins and dyneins. These findings serve to highlight both consistencies with eutherian sperm epididymal maturation (e.g. bias towards protein loss over transit, transfer of proteins via extracellular vesicles) and elements which are likely unique to marsupials (e.g. reduced chromatin stability, potential use of β-oxidation as a major metabolic pathway). This critical information can now be leveraged to further develop ARTs in marsupials.
Project description:Purpose: The goal of this study is to detect differentially expressed genes, among Wild type caput, corpus, and cauda epididymis by RNA sequencing Methods: Caput, corpus, and cauda epidiymal mRNA profiles of 9-month-old wild-type mice were generated by deep sequencing, in triplicate, using Illumina GAIIx. Results: RNA-seq data identified transcripts differentially expressed in caput, corpus, and cauda epididymis. Conclusions: Our results show that the expression of many genes were differentially regulated in caput, corpus, and cauda epididymis.
Project description:Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate with the loss of sperm function during cryopreservation remains unsolved. The present study attempted to clarify this issue evaluating differential changes in the proteome of pig spermatozoa retrieved from the cauda epididymis and the ejaculate, with clear differences in cryotolerance, comparing fresh and frozen-thawed cells. Sperm samples were collected from 10 healthy, sexually mature and fertile boars, and cryopreserved using a standard 0.5 mL straw protocol. Total and progressive motility, viability and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) cauda epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT sperm samples were analyzed using a LC-ESI-MS/MS-based SWATH approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins directly involved in mitochondrial functionality among those quantitatively altered in ejaculated spermatozoa, which would explain the worse post-thaw quality of ejaculated pig spermatozoa.
Project description:Normal epididymis development is the basis of male reproduction, and epididymis is a crucial site where sperm maturation occurs. Although epididymis has been studied through multiple omics methods, combined omics has not been used to present the differential genes in the epididymal cauda of yaks before and after sexual maturity. We here provided the landscape maps of differential genes and proteins obtained through RNA-seq and proteomics technology, and searched for potential key genes, including TGFBI, COL1A1, COL1A2, COL3A1, COL12A1, SULT2B1, KRT19, and NPC2, through integration analysis. In separate and combined enrichment analyses, genes mainly related to cell growth, differentiation and adhesion, sperm maturation, and immune defense were differentially expressed. The involved pathways included extracellular matrix (ECM)–receptor interaction, protein differentiation and absorption, lysosomes, and estrogen signaling pathway. When the sequencing results were verified through qRT-PCR and 4D-PRM, they were found to be consistent. In addition, after the key genes were identified, abnormal expression of these genes was found to possibly cause the retardation of cauda epididymis development and abnormal sperm function in yaks. In conclusion, we provide useful clues for the development of epididymal cauda of yak, sperm maturation, and screening of key genes involved in reproductive regulation of male yak through separate and combined analyses.
Project description:It has been demonstrated that males exposed to adversity prior to conception sire offspring exhibiting abnormal behaviour and neuroendocrine function. Epigenetic factors such as microRNA (miRNA) within sperm may be responsible for driving these effects. Synthetic glucocorticoids (sGC) are a class of drug that are commonly prescribed and may have implications for intergenerational transmission. Therefore, we hypothesized that caput and cauda sperm miRNA profiles will be altered following sGC exposure in guinea pigs. We used miRNA microarray to evaluate the miRNA levels of caput and cauda sperm isolated from guinea pigs exposed to control and water treated with sGC. We identified a subset of miRNAs with low levels in cauda sperm of guinea pigs exposed to sGC.