caa a22 4500 445292334 CHVBK 20180317142532.0 cr unu---uuuuu 170323e20100101xx s 000 0 eng 10.1007/s11626-009-9242-1 doi (NATIONALLICENCE)springer-10.1007/s11626-009-9242-1 In vitro culture of precision-cut testicular tissue as a novel tool for the study of responses to LH [Elektronische Daten] [Andy Laughlin, Thomas Welsh Jr., Charles Love, Dickson Varner, Alan Parrish, David Forrest, Nancy Ing] In vitro culture systems are valuable tools for investigating reproductive mechanisms in the testis. Here, we report the use of the precision-cut in vitro system using equine testicular slices. Testes were collected from immature light breed stallions (n = 3) and cut into slices (mean slice weight = 13.85 ± 0.20mg; mean slice thickness = 515.00 ± 2.33µm) using the precision-cut tissue-slicing method. Four tissue slices were placed on a grid floating on medium in individual vials. After a 1-h preincubation, they were exposed to medium containing ovine luteinizing hormone (oLH) at concentrations of 0, 5, 50, and 500ng/ml for 6h at 32°C. Viability of the tissue was maintained based on histological integrity and lack of appreciable lactate dehydrogenase in the medium. The production and release of testosterone (T) and estradiol-17β (E2) into the medium was measured following in vitro culture. The addition of oLH increased T and E2 at least 400% and 120%, respectively, over the 0-ng oLH control cultures. Testicular gene expression was assessed with in situ hybridization methodology for steroidogenic acute regulatory protein (StAR protein), phosphodiesterase 3B (PDE3B), and outer dense fiber of sperm tails 2 (ODF2) mRNAs. In situ hybridization revealed an oLH concentration-dependent increase in the concentration of StAR protein mRNA in Leydig cells. No differences were observed for the expression of PDE3B or ODF2 genes in seminiferous tubules among treatment groups as expected. These results demonstrate the value of in vitro culture of the precision-cut tissue slices for studies of the regulation of steroidogenesis and gene expression in the stallion testes. The Society for In Vitro Biology, 2009 Organ culture system nationallicence Stallion nationallicence Testis nationallicence StAR protein nationallicence Luteinizing hormone nationallicence Laughlin Andy Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut Welsh Jr. Thomas Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut Love Charles Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 77843, College Station, TX, USA aut Varner Dickson Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 77843, College Station, TX, USA aut Parrish Alan Department of Systems Biology and Translational Medicine, College of Medicine, Texas A&M Health Science Center, 77843, College Station, TX, USA aut Forrest David Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut Ing Nancy Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/1(2010-01-01), 45-53 1071-2690 46:1<45 2010 46 11626 https://doi.org/10.1007/s11626-009-9242-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-009-9242-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Laughlin Andy Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Welsh Jr Thomas Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Love Charles Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Varner Dickson Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Parrish Alan Department of Systems Biology and Translational Medicine, College of Medicine, Texas A&M Health Science Center, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Forrest David Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 700 1- Ing Nancy Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, 77843, College Station, TX, USA aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/1(2010-01-01), 45-53 1071-2690 46:1<45 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer