We examined the expression of the protein and mRNA of the newly cloned isoform of human gonadotrophin-releasing hormone (GnRH-II) in the normal human endometrium during the menstrual cycle. Nested RT-PCR and sequence analysis revealed that two spliced variants of GnRH-II mRNA were expressed during the entire menstrual cycle, with the shorter transcript having a 21 nucleotide deletion in the region coding for GnRH-associated peptide. Using immunohistochemistry, we identified immunoreactive GnRH-II in both stromal and glandular epithelial cells during the entire menstrual phase. The immunostaining intensity was stronger during the early and mid-secretory phase compared with the proliferative and late-secretory phase. A large amount of immunoreactive GnRH-II was localized in the apical pole of the glandular lumen. Our results show that the second isoform of GnRH (GnRH-II) is expressed in the human endometrium during the entire menstrual phase. We also suggest that an increased expression of endometrial GnRH-II peptide, noted during the early and mid-secretory phase, may play an important role in human embryo implantation.