TRANSCRIPTION OF CIRCULAR AND NONCIRCULAR FORMS OF SRY IN MOUSE TESTES

Although its expression in adult testis was immediately apparent, the role for Sry (sex determining region, Y) in testicular function remain s elusive. We have performed transcriptional studies in an effort to e lucidate potential roles of Sry by studying the time and location of i ts transcription in mouse testes. Northern analyses and more sensitive nuclease protection assays detected transcripts in 28-day-old testes and beyond. The highly sensitive technique of reverse transcription po lymerase chain reaction (RTPCR) could not detect Sry expression in 14- day testes when primers for the most conserved portion of the gene, th e high mobility group (HMG) box, were used, but primers for the circul ar form detected Sry transcription at all postnatal stages studied. Th e same HMG box primers were able to detect expression of Sry in XX, Sx r(a) or Sxr(b) testes. This suggested that Sry is expressed in cells o ther than germ cells, which was confirmed with studies on fractionated cells-RTPCR detected transcription of Sry in the highly pure intersti tial cell fraction. However, Leydig cells and a Leydig cell tumor were negative for Sry expression. We performed in situ studies in an attem pt to localize the expression of Sry in the testes. Abundant expressio n of an Sry cross-hybridizing transcript was found in spermatogonia, i n early spermatocytes, and in some interstitial cells with antisense p robes to the HMG box or a more specific, 3' region, whereas the sense probe gave little or no hybridization. It is probable that the circula r transcripts, which are seen in reverse transcriptase positive (RT+) and RT- reactions by PCR because of the RT activity of Taq polymerase, are responsible for the hybridization seen in spermatogonia and sperm atocytes, whereas linear and circular forms are detected later. Thus S ry is expressed in pre- and postmeiotic germ cells and in somatic cell s of the testes. (C) 1994 Wiley-Liss, Inc.