Relaxin-like factor (RLF) mRNA expression in the fallow deer

Employing RT- and RACE-PCR on RNA isolated from testicular tissue, we have cloned the coding cDNA sequence for the RLF, also known as Ins13, of the fa llow deer. The RLF coding sequence consisted of 396 bp encoding a peptide o f 131 amino acids and shared highest homology with bovine, sheep and goat R LF. Northern analysis revealed a single 0.9 kb transcript in the deer testi s. There is only one RLF gene in the deer genome. Nonradioactive in situ hy bridization revealed the Leydig cells to be the sole source for RLF mRNA in the deer testis. In the non-pregnant uterus, RLF transcripts were located in the luminal and glandular epithelium of the endometrium. Within the ovar y of the pregnant doe, follicular theca interns cells and the corpus luteum expressed RLF transcripts. In uteroplacental tissues, luminal and glandula r epithelium, fetal uninucleate and binucleate trophoblast cells (BNC) of t he basic villous trophoblast layer expressed RLF mRNA. BNC located at the a pical trophoblast layer or the tip of the fetal villus were devoid of RLF t ranscripts. Pseudostratified trophoblast cells at the base of fetal villi c oexpressed RLF mRNA and immunoreactive MHC class Ib molecules. (C) 2000 Els evier Science Ireland Ltd. All rights reserved.