| First Author | McCarrey JR | Year | 1992 |
| Journal | Genet Anal Tech Appl | Volume | 9 |
| Issue | 4 | Pages | 117-23 |
| PubMed ID | 1282026 | Mgi Jnum | J:3810 |
| Mgi Id | MGI:52318 | Doi | 10.1016/1050-3862(92)90051-6 |
| Citation | McCarrey JR, et al. (1992) Semiquantitative analysis of X-linked gene expression during spermatogenesis in the mouse: ethidium-bromide staining of RT-PCR products. Genet Anal Tech Appl 9(4):117-23 |
| abstractText | We have used analysis of ethidium-bromide-stained reverse transcriptase-polymerase chain reaction (RT-PCR) products to assess the effects of X-chromosome inactivation during spermatogenesis in the mouse. RT-PCR was performed on total RNA from eight different spermatogenic cell types, including premeiotic spermatogonia, meiotic spermatocytes, and postmeiotic spermatids, to detect transcripts from five different X-linked structural genes (Pgk-1, Zfx, Pdha-1, Hprt, and Phka) and two autosomal genes (Pgk-2 and beta-actin). Relative intensities of ethidium-bromide-stained RT-PCR products representing transcripts from each gene in each cell type were analyzed by densitometry using the Image program (version 1.4, NIH), and normalized against beta-actin values. These results suggest a coordinate inactivation of the X-linked loci at the onset of meiosis, followed by variable rates of decline of corresponding transcript levels reflecting differential mRNA stabilities and/or leaky expression after inactivation. Technically, these results indicate that analysis of ethidium-bromide-stained RT-PCR products can be used to provide a semiquantitative indication of relative levels of specific transcripts in a developing cell lineage without using radioactive probes to quantitate these products. |
