Neurobiología de los péptidos opioides / Neurobiology of opioid peptides

En los últimos 25 años hemos asistido a la formación de nuevas e importantes líneas de investigación en el dominio de las neurociencias, producido por el estudio de los neuropéptidos, los cuales constituyen virtualmente una nueva clase de transmisores químicos. Muchos de ellos pueden funcionar como hormonas endócrinas, parácrinas y amacrínas, además de poseer la cualidad de influir en funciones como las cognocistivas y en los facores de crecimineto. La diversidad bioquímica de las células nerviosas ha revelado la coexistencia, en la misma célula, de neuropéptidos con neurotransmisores clásicos, propiedad que ofrece nuevas perspectivas en la transmisión sináptica. Posiblemente, el grupo de neuropéptidos más grande hasta ahora identificado y estudiado, cuya influencia es decisiva en la homeostasis celular, es el de los péptidos opioides. En la presente revisión hacemos una breve reseña histórica del descubrimiento de las encefalinas, y ofrecemos un panorama general y actualizado sobre los procesos implicados en el metabolismo de los opioides, su síntesis, liberación, tipos de receptores y catabolismo. Para concluir, pasamos revista a las diferentes funciones en las cuales pueden estar implicados. El conocimiento acumulado en dos décadas de estudio sobre las propiedades y funciones de los péptidos opioides aunado a las técnicas modernas en la biología molécular y de diagnóstico clínico, como la tomografía por emisión de positrones y la resonancia magnética nuclear, nos permiten avisorar un panorama prometedor de intervención terapéutica

Thiazide diuretic receptors: autoradiographic localization in rat kidney with [3H]metolazone.

The localization of binding sites for [3H]metolazone, a quinazolinesulfonamide diuretic with thiazide-like actions, was determined by in vitro autoradiography. [3H]Metolazone bound saturably to rat kidney sections incubated in vitro with a dissociation constant (Kd) = 3.4 nM and binding site density = 0.14 pmol/mg of protein. Incubation conditions were used that excluded binding to low affinity sites and carbonic anhydrase. Pharmacological specificity of binding was consistent with labeling of physiologically relevant thiazide diuretic receptors, as identified in previous studies of [3H]metolazone binding to renal membranes. Autoradiographs obtained with tritium-sensitive film demonstrated that binding sites were limited to the renal cortex and were relatively sparsely distributed. Higher resolution autoradiography indicated that [3H] metolazone binding sites were localized in a highly specific manner over short lengths of tubular segments, which by their morphology and distribution most likely represented distal convoluted tubules. In the short sections of tubule that contained receptors, labeling was very dense and appeared to be more prevalent over luminal than peritubular surfaces. The intrarenal distribution of [3H]metolazone binding sites provides further evidence for their identity as thiazide diuretic receptors. These results are consistent with physiological studies demonstrating that the early distal tubule is the location of thiazide-sensitive sodium chloride cotransport.

Receptor-mediated entry of peroxidases into the parasitophorous vacuoles of macrophages infected with Leishmania Mexicana amazonensis.

Leishmania amastigotes, obligatory parasites of macrophages, lodge and multiply within long-lived phagolysosomelike "parasitophorous vacuoles" (PV). The glycoprotein horseradish peroxidase (HRP) was shown, by light and electron microscopic cytochemistry, to enter the PVs of rat in vitro-derived bone marrow macrophages infected with Leishmania mexicana amazonensis. Uptake was obtained both in preinfected macrophages incubated with HRP and in macrophages pulsed with HRP, infected, and further incubated in ligand-free medium. Peroxidase positive and negative PVs could coexist in the same macrophages. Infected macrophages commonly displayed fewer labeled secondary lysosomes than noninfected cells. Lactoperoxidase (LP) was also shown, by light microscopy, to enter the PVs of rat macrophages. Uptake of HRP and of LP was blocked by mannan, supporting the mannose receptor mediated recognition of these ligands. Transfer of HRP to PVs was much less efficient in resident mouse peritoneal macrophages, even at 50 X higher ligand concentrations. Such macrophages expressed negligible mannose receptor function. The efficient mannan-inhibitable uptake of HRP by rat marrow macrophages was confirmed biochemically. Bulk HRP uptake in infected and noninfected cultures was found to be similar. Peroxidases should be useful in further studies of endocytosis by Leishmania-infected macrophages and in the development of lysosomotropic macrophage-targeted drug carriers.