Localization of lipocalin-type prostaglandin D synthase (beta-trace) in iris, ciliary body, and eye fluids.
OBJECTIVE
Prostaglandin (PG) D synthase is present in neural tissues and cerebrospinal fluid (beta-trace). This enzyme belongs to the lipocalin family which consists of transporter proteins for lipophilic substances in the extracellular space. PGD synthase is found in retinal pigment epithelium, from where it is secreted into the interphotoreceptor matrix. The authors have undertaken the localization of this unique enzyme within the tissues and spaces of the anterior segment of the eye.


METHODS
Iris, ciliary body, lens, and aqueous and vitreous humors were collected from adult rats and mice. PGD synthase activity was determined, and the protein was quantified by Western blot analysis and localized immunohistochemically. Finally, in situ hybridization was performed to localize PGD synthase mRNA.


RESULTS
PGD synthase was most abundant in the aqueous and vitreous humors. It was less abundant in tissue cytosolic fractions; these fractions had almost 10-fold as much as their corresponding membrane-bound fractions. Lens tissue had the lowest amount observed. PGD synthase was localized to the epithelial cells of the iris and the ciliary body and to the adjacent extracellular chambers, but PGD synthase mRNA was found only within the epithelial cells. Several glycosylated forms of PGD synthase were also detected.


CONCLUSIONS
PGD synthase was synthesized within the epithelial cells of the iris and the ciliary body and was then secreted into the aqueous and vitreous humors, where it accumulated as an active enzyme.
Gerashchenko
D Y
DY
Department of Molecular Behavioral Biology, Osaka Bioscience Institute, Japan.
Beuckmann
C T
CT
Marcheselli
V L
VL
Gordon
W C
WC
Kanaoka
Y
Y
Eguchi
N
N
Urade
Y
Y
Hayaishi
O
O
Bazan
N G
NG
D000818:Animals; D001082:Aqueous Humor / Q000201:enzymology*; D015153:Blotting, Western; D002924:Ciliary Body / Q000201:enzymology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D017403:In Situ Hybridization; D019746:Intramolecular Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D007498:Iris / Q000201:enzymology*; D007908:Lens, Crystalline / Q000201:enzymology; D054834:Lipocalins; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010857:Pigment Epithelium of Eye / Q000201:enzymology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014822:Vitreous Body / Q000201:enzymology*
D054834:Lipocalins; D012333:RNA, Messenger; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase