Graefe's Arch Clin Exp Ophthalmol (1999) 237:231±240 Springer-Verlag 1999 CLINICAL INVESTIGATION Keith G. Duncan Michele D. Jumper Ralff C. J. Ribeiro Kathy R. Bailey Paul M. Yen Akira Sugawara Ashmi Patel Robert Stern William W. Chin John D. Baxter Daniel M. Schwartz Human trabecular meshwork cells as a thyroid hormone target tissue: presence of functional thyroid hormone receptors Received: 5 February 1998 Revised version received: 2 June 1998 Accepted: 6 July 1998 K.G. Duncan ´ R.C.J. Ribeiro K.R. Bailey ´ J.D. Baxter Metabolic Research Unit, University of California, San Francisco, California, USA K.G. Duncan ´ M.D. Jumper A. Patel ´ D.M. Schwartz ( ) ) Department of Ophthalmology, University of California, San Francisco, 10 Kirkham Street, Box 0730, San Francisco, CA 94143, USA Tel. 001-415-476-1887 Fax 001-415-476-0336 P.M. Yen ´ A. Sugawara ´ W.W. Chin Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA R. Stern Department of Pathology, University of California, San Francisco, California, USA Abstract. l Purpose: To deter- mine whether human trabecular meshwork cells (HTM) are a po- tential target tissue for thyroid hor- mone (3,3©,5-triiodothyronine, T 3 ). l Methods: Cultured HTM were as- sayed for the presence of thyroid hormone receptors (TRs) and retinoid X receptors (RXRs) by reverse-tran- scriptase polymerase chain reaction (RT-PCR) to detected TR and RXR mRNA, and by immunohistochemis- try to detect nuclear TR and RXR proteins. Functionality of the TR was determined by analysis of 125 I-T 3 binding affinity and capacity in HTM nuclear extracts. Effects of T 3 on modulation of hyaluronic acid (HA) levels in HTM were measured as a function of dose and duration of T 3 administration. l Results: Analysis of RT-PCR and immunohistochem- istry demonstrated that cultured HTM expressed TRa1, TRa2, and TRb1 but not TRb2; and RXRa but not RXRb and RXRg isoforms. Satura- tion binding and analysis of 125 I-T 3 to HTM nuclear extracts revealed K d of 57 pM. The number of T 3 binding sites extrapolated from a Scatchard plot was 7.310 10 /mg of HTM nuclear protein extract. T 3 supplementation reduced the concentration of HA in the cell medium by 32±43% com- pared to cells grown in the absence of T 3 . l Conclusions: Cultured HTM express three TR isoforms and one RXR isoform, bind T 3 with an affin- ity similar to that of TR in responsive cells, and modulate their HA pro- duction in response to T 3 . These findings indicate that the human tra- becular meshwork tissue has the ca- pacity to respond to thyroid hor- mones. Introduction There has been interest in the association between prima- ry open-angle glaucoma (POAG) and hypothyroidism since the report by Hertel in 1920 of two patients with both of these disorders [17] in whom systemic thyroid hormone replacement resulted in normalization of the in- traocular pressure (IOP). Numerous subsequent studies addressed this association [4, 7, 8, 14, 16, 27, 35, 38, 45, 47, 51±53], with conflicting conclusions. Supporting an association between hypothyroidism and POAG, Smith et al. used currently accepted methods of evaluat- ing thyroid function and found that 23.4% or 64 patients with POAG were hypothyroid compared to 4.7% of age- matched controls [53]. In patients with newly diagnosed hypothyroidism, Smith et al. observed that 14/25 (56%) had abnormal outflow facility [52]. Systemic replace- ment with thyroid hormone resulted in euthyroid status in all patients and normalization of outflow facility in all but one patient. McDaniel and Besada have also de- scribed one hypothyroid patient whose IOP was reduced with systemic thyroid replacement therapy [34]. Gillow et al., using similar methods of evaluating thyroid func-