TY - JOUR ID - 3446 TI - Localization of Vasa mRNA in testicular tissue of Caspian trout (Salmo caspius) using in situ hybridization JO - Aquatic Physiology and Biotechnology JA - JAPB LA - en SN - 2345-3966 AU - Poursaeed, Samaneh AU - Kalbassi, Mohammad Reza AU - Hassani, Seyedeh Nafiseh AU - Yushizaki, Goro AU - Baharvand, Hossein AD - PhD in Fisheries Science, Department of Fisheries, Faculty of Marine Sciences, Tarbiat Modares University, Noor, Iran AD - Professor in Department of Fisheries, Faculty of Marine Sciences, Tarbiat Modares University, Noor, Iran AD - Assistant Professor in Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran AD - Professor in Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Tokyo, Japan AD - Professor in Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran Y1 - 1970 PY - 1970 VL - 7 IS - 1 SP - 23 EP - 39 KW - Germ Cells KW - In Situ Hybridization KW - Vasa KW - Salmo caspius DO - 10.22124/japb.2019.8176.1184 N2 - In situ hybridization is a powerful tool to localize a specific sequence of nucleic acids in heterogeneous cell population. Therefore, the aim of this study was to apply this technique for localization of Vasa mRNA expression in testicular tissue of Caspian trout (Salmo caspius). In this study, a digoxigenin-labeled RNA probe was synthesized using the plasmid pGEM-T carrying a Caspian trout Vasa cDNA fragment as a template. The RNA labeling was performed through in vitro transcription reaction with DIG labeling mix. For in situ hybridization of tissue sections, Caspian trout testes were fixed and dehydrated. After rehydration and treatment with proteinase K, sections were incubated with hybridization buffer containing Vasa antisense riboprobe for 16h. After washing with SSC solution and incubation with anti-Dig-digoxigenin Antibody, the sections were treated with NBT/BCIP. The results of in situ hybridization using a Vasa antisense riboprobe showed that all developmental stages of male germ cells (spermatogonial stem cells to spermatozoa) expressed this gene, but the expression level was different among germ cells. Therefore, Vasa gene is introduced as a specific marker to identify germ cells and in situ hybridization can be used as a technique to evaluate the localization and expression level of gene mRNA in testicular tissue. These findings may provide important information to help understand the formation and development of germ cells in this valuable species of Caspian Sea. UR - https://japb.guilan.ac.ir/article_3446.html L1 - https://japb.guilan.ac.ir/article_3446_e4e49714f099cf1e87eb91f5d9639732.pdf ER -