Carbonic anhydrases (CAs) plays a critical functional role in the ciliary body and retina for maintenance of microenvironment. With immunohistochemistry using orbital contents from 8 human fetuses (12-16 weeks of gestation), we examined expressions of CAs isozymes-1, 2, 3, 6, 7 9 and 12 and found strong reactivity of CA9 in extra-ocular fibrous tissues in the anterior and posterior eyes. CA9 is known to express in the fetal joint cartilage to maintain pH against hypoxia: actually, in the present specimens, the SO pulley and its tendon was strongly positive for CA9. The CA9-positive anterior fibrous tissues were positive for smooth muscle actin and connected the orbital aspect of the 4 rectus muscle with the palpebral conjunctiva, whereas the posterior tissue was negative for smooth muscle actin and corresponded to the lateral insertion tendon of the orbitalis muscle. The anterior CA9-positve tissues seemed to correspond to the primitive form of the sleeve and pulley system. Any of matrix substances (collagen types I and II, aggrecan, versican, fibronectin, tenascin and hyaluronan) displayed a distribution pattern specific for the CA9-positive fibrous tissues. Therefore, whether or not CA9 was positive in the fibrous tissue seemed not to depend on the tissue components such as the extracellular matrix and intermediate filaments but to suggest a stressful condition such as hypoxia, unsuitable base balance and/or under mechanical stress.