Endothelin A receptor-like immunoreactivity on the basal infoldings of rat renal tubules and collecting ducts.

We investigated the distribution of endothelin A (ET(A)) receptor-like immunoreactivity in the rat kidney using affinity-purified antibodies against amino acid residues 403-417 of the rat ET(A) receptor modified by the multiple antigen peptide complex system. Western blot analysis using the affinity-purified anti-ET(A) antibody detected bands of approximately 47.3 and 64.5 kDa in the rat kidney. By light microscopy, ET(A) receptor-like immunoreactivity was seen in the basal side of the renal tubules and collecting ducts. The most intense immunoreactivity was present in the distal renal tubules and inner medullary collecting ducts. In addition to the basal infoldings, immunoreactive puncta were scattered in the epithelial cells of the renal tubules and collecting ducts. Specimens prepared using the pre-embedding method were examined by electron microscopy, and some immunopositive signals were seen on the basal infodings of the renal tubules and collecting ducts. The lengths of immunopositive cytoplasmic membrane were far longer in the distal tubules and inner medullary collecting ducts than in the proximal tubules and outer medullary collecting ducts. Immunopositive signals were also sometimes observed in the thick portion of Henle's loop, but never in the thin portion. We have not previously detected immunopositive signals on the renal vascular systems with the antibody used here. These results suggest that endothelin acts on the basal infoldings through the ET(A) receptor, particularly in the distal tubules and inner medullary collecting ducts, although involvement of the ET(B) receptor cannot be excluded.

Localization of orexin-A-like immunoreactivity in prolactin cells in the bullfrog (Rana catesbeiana) pituitary.

Immunohistochemical techniques were employed to investigate the distribution of orexin-A-like immunoreactivity in the bullfrog (Rana catesbeiana) pituitary. Orexin-A-immunoreactive cells were scattered throughout the pars distalis. We found that these cells corresponded to the cells immunostained with antiserum against bullfrog prolactin (fPRL). Immunoelectron microscopic analysis indicated that an orexin-A-like substance coexisted with fPRL within secretory granules. Western blot analysis of bullfrog pituitary extract revealed that anti-human orexin-A antiserum labeled two separate bands which were not labeled with anti-fPRL antiserum. The present study has, for the first time, provided evidence of the intragranular colocalization of orexin-A-like and PRL immunoreactivities in the bullfrog pituitary.

Endothelin B receptor-like immunoreactivity in podocytes of the rat kidney.

The distribution of endothelin B receptor (ETBR)-like immunoreactivity in the rat renal glomerulus was investigated using an affinity-purified antibody against a synthetic peptide corresponding to the amino acid residues 425-439 of the rat ETBR. Light microscopy showed ETBR-like immunoreactivity to be localized predominantly near the glomerular blood capillaries. By immunoelectron microscopy using the pre-embedding method, intense immunodeposits indicating ETBR were detected in podocytes, particularly in their foot processes, in contrast with the weak immunoreaction in endothelial cells of the glomerular blood capillaries and in the mesangial cells. In sections stained with the post-embedding method using immunogold particles, positive signals were also found on the plasma membrane of podocyte foot processes as well as the cytoplasm just beneath the cell membrane. These findings suggest that endothelin stimulates ETBR mainly on podocytes, thus resulting in a decrease of the glomerular blood flow and glomerular filtration rates.

Neuropeptide Y-immunoreactive neuronal system and colocalization with FMRFamide in the optic lobe and peduncle complex of the octopus (Octopus vulgaris).

The distribution of neuropeptide Y (NPY)-like immunoreactivity and its colocalization with FMRFamide were investigated in the optic lobe and peduncle complex of the octopus ( Octopus vulgaris) by using immunohistochemical techniques. In the optic lobe cortex, NPY-immunoreactive (NPY-IR) fibers were observed in the plexiform layer, although no NPY-IR somata were observed in the outer or inner granular cell layers. In the optic lobe medulla, NPY-IR somata were seen in the cell islands, and abundant NPY-IR varicose fibers were observed in the neuropil. Most of the NPY-IR structures in the medulla showed FMRFamide-like immunoreactivity. In the peduncle lobe, abundant NPY-IR and FMRFamide-IR (NPY/FMRF-IR) varicose fibers were seen in the basal zone neuropil of the peduncle lobe. In the olfactory lobe, NPY/FMRF-IR varicose fibers were also abundant in the neuropil of the three lobules. NPY/FMRF-IR somata, with processes running to various neuropils, were scattered in the median and posterior lobules. In the optic gland, many NPY/FMRF-IR varicose fibers formed a honeycomb pattern. These observations suggest that NPY/FMRF-IR neurons in the optic lobes participate in the modulation of visual information and that those in the optic gland are involved in the regulation of endocrine function.

Ultrastructure and Atrial Natriuretic Peptide(ANP)-like Immunoreactivity of Cardiocytes in the Larval, Metamorphosing and Adult Specimens of the Japanese Toad, Bufo japonicus formosus: toad/atrial natriuretic peptide(ANP)-like material/immunohistochemistry/immunocytochemistry/RIA.

The distribution of an atrial natriuretic peptide (ANP)-like material in the cardiocytes of larval, metamorphosing, and adult specimens (both breeding and non-breeding) of the toad, Bufo japonicus formosus, was studied immunohistochemically, ultrastructurally and immunocytochemically. Histochemically, ANP-immunoreaction was positive in the atrium and ventricle in stage 33 larvae, while negative in the ventricle in stage 40 larvae. In adult toads, the reaction was stronger in the right than in the left atrium but quite weak in the ventricles, particularly those of non-breeding specimens. Electron microscopy indicated a very small number of secretory granules in the atrial and ventricular cardiocytes of embryos as early as the limb-bud stage (stage 28), and as development proceeded, the number of these granules increased rapidly in atrial but not in ventricular cardiocytes. In metamorphosing animals, a small population of larger granules (200-250 nm) was noted next to those of ordinary size (the median, 110 nm) in the same cell. In adult toads, granules of about 120 nm and 200 nm in median size were found in the same cell. Postembedding immunogold staining consistently indicated ANP-immunoreactivity in these granules in atrial and ventricular cardiocytes. The plasma content of immunoreactive ANP was considerably higher in breeding (20.5 ± 5.9 pg/ml) than in non-breeding toads (5.4 ± 1.7 pg/ml). These results are discussed in relation to presently available data on the physiological role of ANP.