Abstract: 331

CONSTRUCTION AND PRELIMINARY CHARACTERIZATION OF A SERIES OF MOUSE AND RAT TESTIS cDNA LIBRARIES.

John R. McCarrey^{1 *}, Deborah A. O'Brien^{2 *}, Michael K. Skinner^{3 *}
Dept. Genetics; Southwest Foundation for Biomedical Research; San Antonio, TX 78245 ¹
Labs. Repr. Biol. & Depts. Cell Biol. and Anat. & Peds.; University of North Carolina, Chapel Hill, NC 27599 ²
Ctr. Repr. Biol. & Dept. Genetics & Cell Biol.; Washington State University, Pullman, WA 99164 ³

We have constructed a series of 23 cDNA libraries from mouse and rat testicular cells. These include libraries made from whole intact adult testes, seminiferous tubule cells from adult testes, combined populations of primary spermatocytes from 18-day old mouse testes, and isolated populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene spermatocytes, leptotene plus zygotene spermatocytes, juvenile pachytene spermatocytes, adult pachytene spermatocytes, round spermatids, Sertoli cells from 6-, 8-, 17-, and 18-20 day old mice, and peritubular cells from 18-20 day old mice, all recovered from outbred white Swiss (CD-1) mice. We also constructed libraries from whole adult testes from five other lines of mice, C57Bl/6J, C³HeB, BDF-1, Balb/c, and 129/SvJ. Finally, there are two libraries made from populations of Sertoli cells and peritubular cells isolated from testes of 20-day old Sprague Dawley rats. Enzymatic dissociation, followed by gradient separation or plating/lysing techniques were used to prepare populations of specific cell types in purities of 85-98%. cDNAs were synthesized from poly A⁺ mRNA primed with oligo dT and unidirectionally cloned into the lambda Uni-Zap XR expression vector from Stratagene. Primary titers ranged from 2.1 x 10⁵ to 2.9 x 10⁸ plaque forming units, and insert sizes averaged 1.0-1.2 kb. These libraries have been amplified once and submitted to the American Type Culture Collection (ATCC) for distribution to interested investigators. ATCC accession numbers are provided.

    This abstract is being presented on Monday, August 2 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.