Gender-related factors have been attributed to observed differences in the rate of injury to ligaments (e.g., anterior cruciate ligament) between male and female subjects. These differences may be a result of unique regulatory mechanisms within the tissue in response to the sex hormones estrogen and progesterone. These hormones, when bound to specific intracellular receptors (estrogen receptor and progesterone receptor, respectively), modulate gene expression within hormone-responsive tissue. The purpose of this study was to evaluate the expression of the estrogen and progesterone receptors in ligament tissue from male and female rabbits and humans by the sensitive molecular technique of reverse transcription-polymerase chain reaction. Total RNA was extracted from human anterior cruciate ligament tissue and from medial cruciate ligament, anterior cruciate ligament, patellar tendon, and synovium tissue of the New Zealand White rabbit by the newly developed TRIspin method. The total RNA was reverse transcribed and analyzed by polymerase chain reaction to assess the expression of estrogen and progesterone receptors. Our results demonstrate that estrogen and progesterone receptor transcripts are expressed in ligament tissue of male and female rabbits and humans and that alterations in receptor expression occur in ligaments during pregnancy. In the human samples, only a small percentage of the estrogen receptor appeared to be a nonfunctional mRNA splice variant, and the predominant form contained the estrogen-binding domain.