Identification of epididymis-specific transcripts in the mouse and rat by transcriptional profiling.
As part of our efforts to identify novel contraceptive targets in the epididymis we performed transcriptional profiling on each of the 10 and 19 segments of the mouse and rat epididymidis, respectively, using Affymetrix whole genome microarrays. A total of 17 096 and 16 360 probe sets representing transcripts were identified as being expressed in the segmented mouse and rat epididymal transcriptomes, respectively. Comparison of the expressed murine transcripts against a mouse transcriptional profiling database derived from 22 other mouse tissues identified 77 transcripts that were expressed uniquely in the epididymis. The expression of these genes was further evaluated by reverse transcription polymerase chain reaction (RT-PCR) analysis of RNA from 21 mouse tissues. RT-PCR analysis confirmed epididymis-specific expression of Defensin Beta 13 and identified two additional genes with expression restricted only to the epididymis and testis. Comparison of the 16 360 expressed transcripts in the rat epididymis with data of 21 other tissues from a rat transcriptional profiling database identified 110 transcripts specific for the epididymis. Sixty-two of these transcripts were further investigated by qPCR analysis. Only Defensin 22 (E3 epididymal protein) was shown to be completely specific for the epididymis. In addition, 14 transcripts showed more than 100-fold selective expression in the epididymis. The products of these genes might play important roles in epididymal and/or sperm function and further investigation and validation as contraceptive targets are warranted. The results of the studies described in this report are available at the Mammalian Reproductive Genetics (MRG) Database (http://mrg.genetics.washington.edu/).
Johnston
Daniel S
DS
Contraception, Women's Health and Musculoskeletal Biology, Wyeth Research, 500 Arcola Rd, N2312 Collegeville, PA 19426, USA. johnstd2@wyeth.com
Turner
Terry T
TT
Finger
Joshua N
JN
Owtscharuk
Tracy L
TL
Kopf
Gregory S
GS
Jelinsky
Scott A
SA
D000818:Animals; D004822:Epididymis / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods*; D008297:Male; D051379:Mice; D009928:Organ Specificity; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D012313:RNA