ABSTRACT
The process of folliculogenesis requires a tightly regulated series of gene expression that are a pre-requisite to the development of ovarian follicle. Among these genes, follicle-stimulating hormone (FSH) is notable for its dual role in development of follicles as well as proliferation and differentiation of granulosa cells. The post-transcriptional expression of these genes is under the control of microRNAs (miRNAs), a class of small, endogenous RNAs that negatively impact gene expression. This study was carried out to determine the role of several miRNAs including mir-143, let-7a, mir-125b, let-7b, let-7c, mir-21 in follicular development in the mouse. The expression of these RNAs was very low in primordial follicles but these became readily detectable in the granulosa cells of primary, secondary and antral follicles. We show that this expression of some miRNAs (mir-143, let-7a, mir-15b) is under negative control of FSH. Together, these findings suggest that FSH regulates folliculogenesis by a novel pathway of miRNAs.
Subject(s)
Follicle Stimulating Hormone/physiology , MicroRNAs/genetics , Ovary/metabolism , Animals , Base Sequence , Blotting, Northern , DNA Probes , Female , In Situ Hybridization , Mice , Oligonucleotides/genetics , Polymerase Chain ReactionABSTRACT
The aim of this study is to screen the novel gene related to the spermatogenesis. A novel rat testis-specific gene LM23 was identified and characterized by differential display PCR with total RNA from rat type A spermatogonia, pachytene spermatocytes, and round spermatids. LM23 cDNA consists of 1896 base pairs (bp) with a complete open reading frame of 936 bp, and encodes a putative protein including 312 amino acids, which shares no significant homology with any known gene. The sequence of LM23 was submitted to GenBank and the Accession No. was AF492385. Multitissue Northern blot and RT-PCR analysis showed LM23 was specific expression in testis, while its expression was not detected in other tissues. Real-time PCR analysis showed that the expression level of LM23 was highest in spermatocytes and very low in spermatogonia. In situ hybridization revealed strong cytoplasmic positive signal in spermatocytes and weak signal in spermatids and spermatogonia. These results indicated LM23 possessed the testis-specific and stage-specific expression characteristics, and possibly involved in rat spermatogenesis.
Subject(s)
Proteins/isolation & purification , Spermatogenesis/genetics , Testis/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cell Cycle Proteins , Cloning, Molecular , In Situ Hybridization , Male , Molecular Sequence Data , Polymerase Chain Reaction , Proteins/chemistry , Proteins/metabolism , RatsABSTRACT
To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells (BMSCs) at defined stages, subtractive cDNA library was established by means of suppression subtractive hybridization. The BMSCs cultured for 12 and 21 d were used as driver and tester, respectively. A subtract library was successfully constructed and five positive clones were selected from the library. Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank. It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1. RT-PCR was performed to confirm the differentially expressed genes. The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day, while the expression of decorin was only detected in the cells of 21-day.