Testosterone Levels and Development of the Penile Spines and Testicular Tissue during the Postnatal Growth in Wistar Rats

Aim of Study: Gonadal hormones exert a profound influence on the development, structure and function of the sexual organs. The testosterone is one of the androgens that plays an essential role in the development of sexual organs in male mammals. Therefore, the present study was undertaken to evaluate the testosterone levels and developmental pattern of the penile spines and seminiferous tubules during early postnatal life of Wistar rats. Methods and Materials: At 7, 14, 21, 28, 35, 42, 49, 56, 63 and 70 days after birth, penile and testicular tissues of male rats were dissected out and fixed for histological study and plasma testosterone levels were determined using high resolution chromatography. Results: An increase in the number of penile follicles, primarily in the distal region of the penis, was observed from postnatal days 14 to 42, followed by a gradual decrease. Penile spines were absent from birth until the first growth peak, which was observed at 42 postnatal days. Both testicular weight and the area of seminiferous tubules showed gradual increases before achieving their highest values at 42 postnatal days. Similarly, a gradual increase in testosterone levels was detected from day 28, with a peak at 42 postnatal days. Conclusions: These data show a temporal association between the development of the penile spines and testicular tissue with gradual increases in testosterone levels. These results may contribute to a better understanding of the behavioral, hormonal and morphological changes underlying sexual maturation in male rats.

Cloning and Analysis of the Promoter Region of Rat uPA Gene

Objective To clone and analyze the promoter sequence of rat urokinase plasminogen activator protein gene. Methods The genomic DNA was extracted from rat testicular tissue. According to urokinase plasminogen activator, the gene sense primer and antisense primer of uPA gene were designed and synthesized, then Touch-Down PCR were performed. After proper purification, the PCR product was sequenced, analyzed with the promoter prediction software and compared with the DNA sequence of rattuas urokinase plasminogen activator. Results The cloned uPA gene was about 1 572 bp in length, which contained a full open-reading frame with 21 bp in length exons, and the upper region of transcriptional start was 1 551 bp in length which was eucaryon transcriptional control area. The 5＇ UTR had a promoter region including a non-responsive TATA-box. Not only the GC-box binding region was found in this gene, but also active protein I （AP1） and SP1 were seen in other regions. Conclusion A 1 572 bp uPA gene fragment （GenBank accession No.X65651） was obtained from rat genomic DNA library, containing eucaryon transcriptional control area with a promoter region, non-conspicuous TATA-box, GC-box and an extron. A non-responsive TATA-box is located at the upper -30 region.

RNAs in the testicular tissue of patients with non-obstructive azoospermia

Circular RNAs(circRNAs)are highly conserved and ubiquitously expressed noncoding RNAs that participate in multiple reproductionrelated diseases.However,the expression pattern and potential functions of circRNAs in the testes of patients with non-obstructive azoospermia(NOA)remain elusive.In this study,according to a circRNA array,a total of 37881 circRNAs were identified that were differentially expressed in the testes of NOA patients compared with normal controls,including 19874 upregulated circRNAs and 18007 downregulated circRNAs.Using quantitative real-time polymerase chain reaction(qRT-PCR)analysis,we confirmed that the change tendency of some specific circRNAs,including hsa_circ_0137890,hsa_circ_0136298,and hsa_circ_0007273,was consistent with the microarray data in another larger sample.The structures and characteristics of these circRNAs were confirmed by Sanger sequencing,and fluorescence in situ hybridization revealed that these circRNAs were primarily expressed in the cytoplasm.Bioinformatics analysis was used to construct the competing endogenous RNA(ceRNA)network,and numerous miRNAs that could be paired with circRNAs validated in this study were reported to be vital for spermatogenesis regulation.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses indicated that genes involved in axoneme assembly,microtubule-based processes,and cell proliferation were significantly enriched.Our data suggest that there are aberrantly expressed circRNA profiles in patients with NOA and that these circRNAs may help identify key diagnostic and therapeutic molecular biomarkers forNoA patients.