title: FK506 blocks activation of the intrinsic caspase cascade after optic nerve crush.
abstract: Retinal ganglion cells die by apoptosis after optic nerve crush. FK506 has been shown to be neuroprotective in this model but the mechanism(s) by which it exerts these actions remains unknown. We and others have shown that caspase 9 is cleaved in the retina in other injury models and we hypothesized that the neuroprotection observed with FK506 was mediated by interference with caspase 9 activation. The present study examined the cellular localization of caspase 9 cleavage after intraorbital optic nerve crush in rats, the time course of caspase 9 cleavage after optic nerve crush and the ability of orally administered FK506 to block caspase 9 cleavage after optic nerve crush. We show by immunohistochemistry that cleaved caspase 9 is present in retinal ganglion cells (identified by prior backlabelling) after optic nerve crush. Immunoblot analysis showed that caspase 9 cleavage is significantly elevated 5 and 8 days after optic nerve crush. We show that orally administered FK506 reaches the retina and is pharmacologically active in retinal tissue. Furthermore, the oral administration of FK506 5 mg kg(-1) day(-1) blocks the cleavage of caspase 9 at both time points. These data suggest that caspase 9 activation may play an important role in retinal ganglion cell death following optic nerve crush and that the neuroprotection seen with FK506 may be mediated by interfering with the activation of caspase 9.
authors.affiliation: Howe Laboratory of Ophthalmology, Massachusetts Eye and Ear Infirmary, Harvard Medical School, 243 Charles Street, Boston, MA 02114, USA. cynthia_grosskreutz@meei.harvard.edu
authors.forename: Cynthia L
authors.initials: CL
authors.lastname: Grosskreutz
authors.forename: Virve A
authors.initials: VA
authors.lastname: Hänninen
authors.forename: Mina B
authors.initials: MB
authors.lastname: Pantcheva
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Nathaniel R
authors.initials: NR
authors.lastname: Poulin
authors.forename: Adam P
authors.initials: AP
authors.lastname: Dobberfuhl
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D019703:Calcineurin / Q000378:metabolism; D053453:Caspase 9; D061945:Caspase Inhibitors; D020169:Caspases / Q000032:analysis / Q000378:metabolism; D003864:Depression, Chemical; D004789:Enzyme Activation / Q000187:drug effects; D007150:Immunohistochemistry / Q000379:methods; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D008297:Male; D009409:Nerve Crush; D020221:Optic Nerve Injuries / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D051381:Rats; D011914:Rats, Inbred BN; D012160:Retina / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000201:enzymology*; D016559:Tacrolimus / Q000627:therapeutic use*; D013997:Time Factors
chemical_list: D061945:Caspase Inhibitors; D007166:Immunosuppressive Agents; D019703:Calcineurin; C506435:Casp9 protein, rat; D053453:Caspase 9; D020169:Caspases; D016559:Tacrolimus