The alpha 1, alpha 2, and alpha 3 chains of collagen VI and mRNAs for these chains were localized in the female mouse reproductive tract by immunofluorescence and in situ hybridization. High levels of collagen VI protein and mRNAs were present in the endometrium and myometrium of the uterus up to Day 4.5 of pregnancy. After embryo implantation, reduction in collagen VI protein within the decidualizing endometrium correlated with significantly reduced steady-state levels of alpha 1(VI), alpha 2(VI), and alpha 3(VI) mRNAs, indicating either transcriptional down-regulation of collagen VI gene expression or decreased stability of transcripts. High levels of alpha 1(V1) and alpha 2(VI) mRNAs, but not alpha 3(VI) mRNA, in cells surrounding the uterine epithelium in the mesometrial region did not correlate with deposition of collagen VI protein in this region. These data are consistent with an important role of alpha 3(VI) in assembly of collagen VI heterotrimers. However, distinct immunostaining with antiserum to alpha 2(VI) chains in the extracellular matrix immediately beneath the uterine epithelium may indicate that alpha 2(VI) chains are deposited without the alpha 1(VI) or alpha 3(VI) collagen chains. No collagen VI protein or mRNAs were detected in any tissue layers of the embryo on Days 5.5 or 6.5 of gestation.