RESUMEN
The activity of angiotensin I-converting enzyme (ACE, EC 3.4.15.1), measured using Hip-His-Leu as substrate, was determined in the developing chick retina, and in monolayer and aggregate cultures of embryonic retinal cells. ACE specific activity in chick retinal homogenate increased 86-fold from embryonic day 13 until the 7th post-hatching day. The development of ACE activity occurred in parallel with that reported for synapse and photoreceptors. ACE activity expression in aggregates, but not in monolayer culture, was similar to that observed in the developing retina in ovo. At culture, day 13, ACE specific activity was 11.8-fold higher in the aggregate than in the dispersed cell culture, and was comparable to that in a 21-day-old embryonic intact retina. Our results suggest that histotypic association of retinal cells during development may be an important event controlling the expression of ACE activity in the CNS.
Asunto(s)
Desarrollo Embrionario y Fetal , Peptidil-Dipeptidasa A/metabolismo , Retina/enzimología , Animales , Células Cultivadas , Embrión de Pollo , Enalaprilato/farmacología , Inhibidores Enzimáticos/farmacología , Oligopéptidos/metabolismo , Peptidil-Dipeptidasa A/fisiología , Retina/embriologíaRESUMEN
The time course of dipeptidase activity and the effect of p-mercuribenzoate (PCMB) on the subestimation of the fluorometric determination of angiotensin-converting enzyme (ACE, EC 3.4.15.1) during development was studied. ACE and dipeptidase activities were measured fluorometrically in homogenates of the developing chick retina using Hip-His-Leu and His-Leu as substrates, respectively, both either in the presence or in the absence of 1 mM PCMB. ACE activity was inhibited by captopril (IC50 1.7 nM), MK 422 (IC50 4.8 nM), BPP9a (IC50 0.25 microM) and BPP5a (IC50 1.2 microM), thus suggesting that avian retinal ACE catalytically resembles the mammalian enzyme. Dipeptidase activity varied 3.4-fold throughout development, leading to a large and variable (28-83%) subestimation of ACE activity during chick retina ontogenesis. PCMB (1 mM) inhibited 67-94% dipeptidase activity during development, thus greatly reducing any subestimation of ACE activity determination during the development of the chick retina.