Project description:Cryptorchidism in the orl rat is associated with muscle patterning defects in the fetal gubernaculum and altered hormonal signaling
Project description:Insl3 is a testis-derived hormone that induces growth and differentiation of the fetal gubernaculum. The goal of this study was to identify genes showing altered expression in fetal gubernaculum following Insl3 exposure.
Project description:During male development, the testes move from a high intraabdominal position and descend into the scrotum. The gubernaculum, an inguinoscrotal ligament connecting the testis to the lower abdomen, is believed to play a critical role in this process. The first stage of testicular descent is controlled by INSL3, insulin like3 hormone, produced in testicular Leydig cells. Deletion of Insl3 or its receptor, Rxfp2, in mice causes cryptorchidism. We produced Cre/loxP regulated shRNA transgenic mice targeting RXFP2 expression. We have shown that the transgene was able to reduce Rxfp2 gene expression and thus behaved as a hypomorphic allele of Rxfp2. Variable degrees of uni-and bilateral cryptorchidism was detected in males with the activated shRNA transgene on an Rxfp2+/- background. Conditional suppression of Rxfp2 in the gubernaculum led to cryptorchidism. Gene expression analysis of a mutant cremasteric sac using Illumina microarrays indicated abnormal expression of a significant number of genes in Wnt/β-catenin and Notch pathways. We have demonstrated profound changes in the expression pattern of β-catenin, Notch1, desmin, and androgen receptor (AR) in Rxfp2-/- male embryos, indicating the role of INSL3 in proliferation, differentiation, and survival of specific cellular components of the gubernaculum. We have shown that INSL3/RXFP2 signaling is essential for myogenic differentiation and maintenance of AR-positive cells in the gubernaculum. Males with the deletion of β-catenin or Notch1 in the gubernacular ligament demonstrated abnormal development. Our data indicates that β-catenin and Notch pathways are potential targets of INSL3 signaling during gubernacular development.
Project description:This study was designed to provide additional insight into testicular hormone production and responsiveness in the orl strain and complement ongoing efforts to characterize the genetic basis of cryptorchidism in this isolated rat colony. Gubernacula were harvested from Long Evans and ORL rat embryos at gestational day 17 and 19. RNA was purified from litter-based organ pools. 26 RNA samples were processed and hybridized to GeneChips. Please note that a mix-up of the samples gubernaculum-GD19-a and -b appears to have occurred as data from these samples clustered with the opposite strain than expected in hierarchical clustering analysis. Thus the strain information was annotated as 'unknown'.
Project description:During male development, the testes move from a high intraabdominal position and descend into the scrotum. The gubernaculum, an inguinoscrotal ligament connecting the testis to the lower abdomen, is believed to play a critical role in this process. The first stage of testicular descent is controlled by INSL3, insulin like3 hormone, produced in testicular Leydig cells. Deletion of Insl3 or its receptor, Rxfp2, in mice causes cryptorchidism. We produced Cre/loxP regulated shRNA transgenic mice targeting RXFP2 expression. We have shown that the transgene was able to reduce Rxfp2 gene expression and thus behaved as a hypomorphic allele of Rxfp2. Variable degrees of uni-and bilateral cryptorchidism was detected in males with the activated shRNA transgene on an Rxfp2+/- background. Conditional suppression of Rxfp2 in the gubernaculum led to cryptorchidism. Gene expression analysis of a mutant cremasteric sac using Illumina microarrays indicated abnormal expression of a significant number of genes in Wnt/β-catenin and Notch pathways. We have demonstrated profound changes in the expression pattern of β-catenin, Notch1, desmin, and androgen receptor (AR) in Rxfp2-/- male embryos, indicating the role of INSL3 in proliferation, differentiation, and survival of specific cellular components of the gubernaculum. We have shown that INSL3/RXFP2 signaling is essential for myogenic differentiation and maintenance of AR-positive cells in the gubernaculum. Males with the deletion of β-catenin or Notch1 in the gubernacular ligament demonstrated abnormal development. Our data indicates that β-catenin and Notch pathways are potential targets of INSL3 signaling during gubernacular development. Total RNA obtained from the cremasteric sac of cryptorchid Tg(shRxfp1) males compared to the wild-type control cremasteric sac.
Project description:Insl3 is a testis-derived hormone that induces growth and differentiation of the fetal gubernaculum. The goal of this study was to identify genes showing altered expression in fetal gubernaculum following Insl3 exposure. The number of biological replicates processed initially were six controls, five 10 nM Insl3, and six 100 nM Insl3. Following data collection, two 10 nM Insl3 samples (biological replicates 4 and 5) were deemed to be outliers based upon Principal Component Analysis and a relatively low yield of biotin-labeled cRNA. The two outlier samples were not included in the statistical analysis.
Project description:This dataset contains high-throughput RNA sequencing (RNA-seq) data generated from gubernaculum and testis samples of wild-type (WT), cetacean INSL3 knock-in (KI), and INSL3 knockout (KO) mice at selected postnatal developmental stages. The purpose of the study was to investigate the molecular consequences of replacing the endogenous mouse Insl3 coding sequence with the cetacean INSL3 ortholog, and to compare transcriptomic profiles among WT, KI, and KO mice to elucidate the functional role of INSL3 in testicular descent and cryptorchidism. RNA-seq was performed on gubernaculum tissue at postnatal day 8 (P8) and testis tissue at P8, P23, and P40 (with some groups excluded due to degeneration or non-specific damage). The dataset supports integrative analyses of developmental stage–specific and genotype–specific gene expression changes in reproductive tissues.