Project description:Hezuo pigs are known in China for early sexual maturity. In this study, we obtained small RNA-Seq data from testicular tissues of 30-day-old and 120-day-old Hezuo and Landrace pigs. Through comparative analysis of their differences, we searched for some miRNAs related to the regulation of precocious sexual traits in male cooperating pigs and revealed their molecular regulatory mechanisms, which can then provide references for the diagnosis and treatment of precocious sexual diseases in humans and for accelerating the breeding of high-fertility animals.
Project description:Postnatal human male gonad development and function are known to involve many genes and pathways but our understanding of genome-wide developmental stage-specific and cell type-specific gene expression is far from complete. Integration of testicular and somatic data could elucidate regulatory mechanisms specifically controlling spermatogenesis and may yield insight into certain reproductive pathologies. Please note: AdMinus means that there is no Ad spermatogonia in the corresponding testicular biopsies of cryptorchid children. AdPlus means that Ad spermatogonia are present in the corresponding testicular biopsies of cryptorchid children. JS refers to Johnsen score.
Project description:Hezuo pigs are known in China for early sexual maturity. In this study, we obtained transcriptome data from testicular tissues of 30-day-old and 120-day-old Hezuo and Landrace pigs. Through comparative analysis of their differences, we searched for some genes related to the regulation of precocious sexual traits in male cooperating pigs and revealed their molecular regulatory mechanisms, which can then provide references for the diagnosis and treatment of precocious sexual diseases in humans and for accelerating the breeding of high-fertility animals.
Project description:Pro-spermatogonia (SG) serve as the gateway to spermatogenesis. Using single-cell RNA sequencing (RNAseq), we studied the development of ProSG, their SG descendants, and testicular somatic cells, during the perinatal period in mice. We identified both gene and protein markers for 3 temporally distinct ProSG cell subsets, including a migratory cell population with a distinct transcriptome from the previously defined T1- and T2-ProSG stages. This intermediate (I)-ProSG subset translocates from the center of seminiferous tubules to the spermatogonial stem cell (SSC) “niche” in its periphery soon after birth. We identified 3 undifferentiated SG subsets at postnatal day 7, each of which express distinct genes, including transcription factor and signaling genes. Two of these subsets have the characteristics of newly emergent SSCs. We also molecularly defined the development of Sertoli, Leydig, and peritubular myoid cells during the perinatal period, allowing us to identify candidate signaling pathways acting between somatic and germ cells in a stage-specific manner during the perinatal period. Our study provides a rich resource for those investigating testicular germ and somatic cell developmental during the perinatal period.
Project description:Smooth-muscle-like peritubular cells make up the wall of semniferous tubules of men. These human testicular peritubular cells (HTPC) have been shown to fulfill different roles. They transport sperm, secrete various factors like GDNF (glial cell line derived neurotrophic factor) and are immunologically active. They might contribute to spermatogonial stem cell niche altering and testicular ageing. Previous studies were limited regarding heterogeneity (due to lifestyle, age, medical history etc.) and accessibility of HTPCs. To circumvent this problem a cellular primate model should be established. The proteome of 6 MKTPCs of individual healthy and young (2 or 3 years) donors was assessed to investigate the suitability as a model.
Project description:The aim of this study was to identify differentially expressed genes and pathways in longissimus dorsi (LD) of pigs at 40 and 70 d of gestation (stages encompassing the transition from primary to secondary fiber formation) in U.S. commercial crossbred pigs (Yorkshire x Landrace) and Brazilian native Piau pigs. We confirmed the expression patterns for a subset of genes by qRT-PCR. Pathway analysis revealed functionally related genes, and indicated commonalities and differences between the breed types and developmental ages evaluated. Results from qRT-PCR analysis confirmed the expression patterns observed on the array for most of the genes tested (85%). This study reveals transcriptional profiles in LD at 40 and 70 d gestation for commercial and Piau pigs, which helps elucidate phenotypic differences between these breed types.
Project description:The aim of this study was to identify differentially expressed genes and pathways in longissimus dorsi (LD) of pigs at 40 and 70 d of gestation (stages encompassing the transition from primary to secondary fiber formation) in U.S. commercial crossbred pigs (Yorkshire x Landrace) and Brazilian native Piau pigs. We confirmed the expression patterns for a subset of genes by qRT-PCR. Pathway analysis revealed functionally related genes, and indicated commonalities and differences between the breed types and developmental ages evaluated. Results from qRT-PCR analysis confirmed the expression patterns observed on the array for most of the genes tested (85%). This study reveals transcriptional profiles in LD at 40 and 70 d gestation for commercial and Piau pigs, which helps elucidate phenotypic differences between these breed types. This study utilized the Swine Protein-Annotated Oligonucleotide Microarray which contains 20,400 70-mer oligonucleotides (http://www.pigoligoarray.org). Total RNA was isolated from fetuses obtained from gilts at each gestational age (n=3 crossbred gilts; n=4 Piau gilts) and RNA from 3 fetuses per litter was pooled. Samples were evaluated with a connected loop design using 13 slides such that six breed comparisons and seven age comparisons were performed. Fluorescence intensity data was LOESS normalized and analyzed with a mixed model.
Project description:The aim of the study was to identify in vivo spermatogonial gene expression within the context of their biological niche. Identification of spermatogonial genes was done by t-testing on the replicates of Score 3 (Spermatogonia) and Score 2 (SCO) testicular biopsies. 3 biological replicates with spermatogonial presence in the tubuli seminiferi, 5 biological replicates with Sertoli-cell-only syndrome.
Project description:PIWI-interacting RNAs (piRNAs) have been identified in mammalian germline initially and later also found to be aberrantly expressed in various cancers. However, their functions in cancer is not very clear. Here, we studied the role of piR-36249, a member of the piRNA family, in testicular cancer. We show that piR-36249 is significantly downregulated in testicular cancer tissues comparing to tumor-adjacent tissues. Functional studies demonstrate that piR-36249 inhibits testicular cancer cell proliferation, migration and invasion. Mass Spectrometry (MS) analysis of piR-36249 pull-downed proteins reveals that piR-36249 interacts with ATP-dependent DNA/RNA helicase (DHX36). Dual-luciferase assays shows that piR-36249 binds to the 3'UTR of 2'-5'-oligoadenylate synthetase 2 (OAS2) mRNA. OAS2 has been shown in literature as a tumor suppressor modulating the occurrence and development of some tumors, such as colorectal cancer and breast cancer. In this study, OAS2 knockdown also promotes testicular cancer cell proliferation and migration. Furthermore, RNA immunoprecipitation assay reveals that DHX36 can also bind to OAS2 mRNA, and knockdown of DHX36 increases OAS2 mRNA yet downregulates its protein, indicating the enhancing OAS2 protein expression level effect of DHX36. All these data suggest that piR-36249, together with DHX36, functions in inhibiting testicular cancer cells proliferation, migration and invasion by upregulating OAS2 protein, and piR-36249 may serve as a potential predictor for the prognosis of testicular cancer.