Molecular and biochemical analysis of ceruloplasmin expression in rabbit and rat ciliary body.
OBJECTIVE
To verify the capability of rabbit and rat ciliary body to synthesize and secrete ceruloplasmin.


METHODS
Isolated ciliary body (CB) was cultured in the presence of [35S]-methionine, and the incubation medium was processed for immunoprecipitation. Total RNA from CB was processed for RT-PCR, and the amplification products were sequenced. Also, sections of CB were immunostained for the localization of ceruloplasmin.


RESULTS
A labeled peptide, having a molecular weight of about 135 kDa, the expected size of ceruloplasmin, was immunopurified in the incubation media from both animal species. The RT-PCR and sequencing experiments detected the presence of ceruloplasmin mRNA in rat samples. Both layers of rabbit and rat ciliary epithelium (CE) exhibited ceruloplasmin reactivity after immunohistochemical processing.


CONCLUSIONS
Taken altogether, these results indicate the CB, particularly its epithelium, as one of the possible sources of the ocular intrinsic ceruloplasmin.
Bertazolli-Filho
Rubens
R
Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, USP, Ribeirão Preto, Brazil.
Laicine
Eduardo Miguel
EM
Haddad
Antonio
A
Rodrigues
Maria Laura Pinto
ML
D000818:Animals; D002570:Ceruloplasmin / Q000235:genetics* / Q000378:metabolism*; D002924:Ciliary Body / Q000201:enzymology*; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D008297:Male; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014168:Transferrin / Q000235:genetics
D012333:RNA, Messenger; D014168:Transferrin; D002570:Ceruloplasmin