Project description:Nitrogen starvation drives meiosis in yeasts1, while retinoic acid (RA) is required for mammalian meiosis through the activation of its germline-specific target, stimulated by retinoic acid gene 8 (Stra8).Here, by using single-cell transcriptome analysis of wild-type and Stra8-deficient testes, our data revealed that the expression of major metabolic genes are downregulated during germ cell entry into meiosis. Recently, we collected mouse testicular cells from wild-type and Stra8-deficient mice at postnatal day 12 (P12) and P21 for single cell RNA-sequencing (scRNA-seq). Upon scRNA-seq, we obtained a total of 23,470 cells from these samples. Using stringent quality control, 22,839 cells were selected for further analysis. All cells, including both wild-type and Stra8-deficient testicular cells, were pooled together to perform clustering analysis, which revealed 6 major cell clusters based on their distinct gene expression patterns. We annotated these six clusters by using known marker genes, including spermatogonia (SPG; Stra8; 3,941 cells), spermatocyte (SPC; Syce1; 4,924 cells), spermatid (STD; Acrv1; 710 cells), Sertoli cell (SC; Sox9; 9,488 cells), Myeloid (MYD; Acta2; 3,531 cells), and Leydig Cells (LY; Star; 245 cells). There is an increase in the percentage of spermatogonia in Stra8-deficient testes at both P12 (28% versus 14% in wild-type) and P21 (17% versus 10% in wild-type). At P21, while spermatocytes are abundant and spermatid are formed in wild-type testes, there are only a few spermatocytes (0.2% versus 73% in wild-type), and spermatids are absent in Stra8-deficient testes. Signature genes are enriched for GO terms for distinct biological function and metabolic pathways in different cell types.
Project description:Ip6k1 knockout male mice are infertile and display a delay in the first wave of spermatogenesis. To understand the underlying basis of this delay, we compared the gene expression profiles of whole testes from 17 days postpartum (17dpp) and 26 days postpartum (26dpp) Ip6k1 wild type (Ip6k1+/+) and Ip6k1 knockout (Ip6k1-/-) mice. We observed deregulation of several biological processes in Ip6k1-/- testes compared with Ip6k1+/+ testes.
Project description:Samples from wild-type C57BL/6 mice gavaged with 100 mg kg-1 of PheCA, SerCA, TCA, or mock control for 13 days and then sacrificed for sampling on day 14.
Project description:Comparative analysis of renal gene expression between wild type and mice deficient in IL-17 receptor signaling following disseminated candidiasis. The hypothesis to be tested in the present study was that there should be a differential renal gene expresion in the absence of IL-17 receptor signaling following disseminated candidiasis. Wild type and IL-17RA-/- mice were systemically infected with Candida albicans and 48 hour post infection renal gene expression was analyzed.
Project description:In order to understand whether sole Cre recombinase expression has an effect on stress signalling pathways and the peroxisomal or other subcellular compartments, we have analyzed total RNA isolated from Sertoli cell cultures of anti-Müllerian-hormone (AMH)-Cre/wild-type and wild-type/wild-type C57BL/6J mouse testes using microarrays. The Sertoli cells were isolated from individual 14 day-old mice and were cultivated for 7 days. Total RNA was isolated from the Sertoli cell culture (for AMH-Cre/WT 5 mice and WT/WT 3 mice) and the same genotypes were pooled together. For the microarray analysis two technical replicates for each genotype were generated.
Project description:Findings from gene expression studies can help identify relevant biological pathways. RNA from three pairs of testes from wild-type and pcd3J-/- mice at postnatal day 18 was subjected to a genome-scale gene expression analysis using the Affymetrix GeneChip Mouse Gene 1.0 ST Array, which includes approximately 28,000 genes. RNA from postnatal day 18 was used to minimize the changes in gene expression that might occur in response to pathological changes in the pcd3J-/- mice, because postnatal day 18 was the earliest time point at which structural differences between testes from wild-type and pcd3J-/- mice were observed via histopathology.Three pcd3J-/- mutant mice and three corresponding wild-type (pcd3J+/+) littermates at postnatal day 18 were used for the microarray analyses.
Project description:We report the effect of Morc2b inactivation on the transcriptome of testes at postnatal day 12 (PND12). We find 88 differentially expressed genes between wild type and Morc2b-/- testes
Project description:Male CD-1 mice (N=8) were intraperitoneally infected by culture-derived Trypanosoma cruzi trypomastigotes of the Brazil strain. One group (N=4) was treated with an antitrypanosomal drug (nifurtimox) on day 74 post-infection (chronic phase) for 20 days. A group of uninfected mice (N=3) was also included as control. All groups (N=11) were enthanized on day 110 post-infection, testes were collected and subjected to 2D-LC-MS/MS analysis. Label-free quantitation revealed that the testes of T. cruzi-infected mice had an altered proteome pattern relative to treated mice and uninfected controls. The analysis also demonstrated that the left and right testes had different proteomic profiles in infected animals.
Project description:The neocortex and striatum were harvested from conditional Met null male mice (Metfx/fx/nestincre) and wild type littermates at postnatal day 14. Synaptosomes were generated and prepared from each region for quantitative proteomic analyses using isobaric tags for relative and absolute quantitation (iTRAQ).