Control of the actomyosin network plays a role in regulating the movement of aqueous humor through the anterior segment of the eye. Receptors that could control its activity are unknown. In this study, we show that all four members of the syndecan family, which can regulate the actomyosin network, are present within the anterior segment. In both sections of human anterior segments and cultures of human trabecular meshwork (HTM), Schlemm's canal (HSC) and the ciliary muscle (HCM) cells from the anterior segment, syndecans-3 and -4 were the predominant family members. They were widely distributed throughout the anterior segment. Syndecan-3 within the anterior segment was a novel, recently described variant 55 kDa form. Low levels of syndecans-1 and -2 were also observed in situ and in all three cultures. Their expression was weaker and more localized than that observed for syndecans-3 and -4. Staining for syndecan-1 in HCM cultures was variable. In HTM and HSC cultures, syndecan-2 also co-distributed with fibronectin, laminin and type IV collagen suggesting that it was shed and associated with the extracellular matrix. Western blots supported this idea and showed syndecan-2 ectodomains in lysates from anterior segments.