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T Isayama

Publications and source records attributed to T Isayama.

45 records · Page 3Linked to original sources

Synaptic analysis of amacrine cells with neuropeptide Y-like immunoreactivity in turtle retina.

Neuropeptide Y-like immunoreactivity has been localized previously within three classes of amacrine cells in the turtle retina. We have used the avidin-biotin with horseradish peroxidase technique to label these neurons for examination at the ultrastructural level to answer the following questions. Where are the synaptic contacts of these neurons made? What types of neurons are involved pre- and postsynaptically? What is the intracellular distribution of the immunoreactivity? Processes with neuropeptide Y-like immunoreactivity were located primarily within three regions of the inner plexiform layer: stratum 1, stratum 3, and at the border between strata 4 and 5. In all three regions the processes with neuropeptide Y-like immunoreactivity received synaptic contacts from both unlabeled amacrine and bipolar cells, but the majority of the synaptic input in all three regions was from unlabeled amacrine cells. Processes with neuropeptide Y-like immunoreactivity were presynaptic to unlabeled amacrine cells in all three regions, but also formed contacts onto unlabeled bipolar cells in the region between strata 4 and 5. The immunoreactivity within these cells gave rise to a diffuse reaction product that was distributed throughout the cytoplasm and within large vesicles. This localization of neuropeptide Y-like immunoreactivity within large vesicles suggests that this peptide may play a neuromodulatory role. Such a role would be consistent with previous studies of neuropeptides in the turtle retina.

Animals↗

Neuropeptide Y-immunoreactive amacrine cells in the retina of the turtle Pseudemys scripta elegans.

Antiserum directed against neuropeptide Y selectively labeled certain amacrine cells in the turtle retina. The cell types, sizes, dendritic stratification, regional distribution, and degrees of immunolabeling were examined. The results indicated that three morphologically distinct cell types were labeled: types A, B, and C. Computer rotation of digitized data from camera lucida drawings was used to study dendritic stratification. The type A somata were large (11.5 micron in diameter), well-stained, and located in the third tier of the inner nuclear layer. Type A somata gave rise to well-stained processes which arborized within the inner plexiform layer in strata 1 and 3 and at the border between strata 4 and 5. Processes in stratum 1 were sparse and delicate with small boutons. Processes in stratum 3 were numerous and often coarse, with many large and small boutons. At the border between strata 4 and 5 the processes were frequently numerous but slender, with many small boutons. Occasional immunolabeled processes were found in the ganglion cell layer. The somata of the type B cells were smaller (9.0 micron in diameter) and gave rise to single labeled processes which descended into the inner plexiform layer and divided quickly into two secondary processes. These secondary processes gave rise to lightly labeled dendritic fields which arborized primarily in strata 2 and 4. The type C cells were usually observed at the periphery of the retina and had large somata (12.0 micron in diameter) with simple, but very elongated, dendritic arborizations in strata 1, 4, and 5. Observations also showed that type A and B cells were often found in close proximity to each other and suggested that dendrites of these cells made contact with each other. The labeled neurons were distributed relatively evenly throughout the retina except for the visual streak where they were sparse. This study indicates that neuropeptide Y-like immunoreactivity is found in more than one anatomically distinct class of amacrine cells in the turtle retina.

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Ganglion cells in the turtle retina contain the neuropeptide LANT-6.

This study investigated the presence of the neurotensin-related hexapeptide, LANT-6, in retinal ganglion cells and their central projections in the turtle Pseudemys scripta elegans. Immunocytochemical techniques demonstrated that many of the cells in the ganglion cell layer of the turtle retina could be labeled with an antiserum specific for LANT-6. Radioimmunoassay and chromatographic analysis confirmed the presence of LANT-6-related peptides in retina, as well as brain. Several molecular forms of LANT-6 were observed, some larger than LANT-6. Characterization of the cells labeled in the ganglion cell layer in terms of their cell body size and their dendritic arborization patterns revealed that at least 6 specific LANT-6-positive cell types were present in the ganglion cell layer. Morphologically, the LANT-6-positive cells strongly resembled turtle ganglion cells, as previously described. In addition, two other lines of evidence supported this interpretation. First, double-label studies were performed in which retinal ganglion cells projecting to the tectum were retrogradely labeled by HRP injected into the tectum (using a cobalt chloride color-modified DAB reaction product) and immunocytochemically labeled with DAB using the antiserum against LANT-6. These double-label studies revealed that many of the LANT-6-positive cells in the ganglion cell layer in the portion of the retina labeled retrogradely by the HRP injection did project to the tectum. Within the retrogradely labeled portion of the retina, LANT-6-positive cells that were not labeled retrogradely, as well as neurons labeled retrogradely that did not contain LANT-6 were also observed. Second, the central projections of LANT-6-positive cells of the ganglion cell layer were examined by studying the effects of monocular enucleation on the distribution of LANT-6-positive fibers in the central projection targets of the turtle retina. Two to 8 weeks after enucleation, a substantial reduction in LANT-6-positive fibers was observed in all retinal target areas contralateral to the enucleated eye. Radioimmunoassay and chromatographic studies confirmed the presence of LANT-6-related peptides in the turtle brain and corroborated the reduction of LANT-6 observed in the contralateral tectum following monocular enucleation. Previous studies have demonstrated that LANT-6-related material is present in cells of the ganglion cell layer in a variety of vertebrates. The present results indicate that LANT-6 is in ganglion cells and that it may play a role in neurotransmission between retinal ganglion cells and their central target areas.

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Intermediate filaments of myofibroblasts. Immunochemical and immunocytochemical analyses.

We generated a monoclonal anti-vimentin antibody, VIM-1, by mouse hybridoma technique, using an established myofibroblast line as a whole cell immunogen. The presence of vimentin polypeptides in the cultured myofibroblasts was confirmed by SDS-polyacrylamide gel electrophoresis and immunoblotting. By light microscopic immunocytochemistry, myofibroblasts in cultures as well as in frozen tissue sections showed a strong reaction with the anti-vimentin antibody, whereas these cells lacked either detectable desmin or cytokeratin. Our results support the fibroblastic origin of myofibroblasts. Immunoelectron microscopic study with ferritin-ABC technique demonstrated that VIM-1 reacted exclusively with 10-nm intermediate filaments, while other cellular structures revealed uniformly negative reaction against the antibody.

Antibodies, Monoclonal↗

Malignant fibrous histiocytoma. Evidence of perivascular mesenchymal cell origin immunocytochemical studies with monoclonal anti-MFH antibodies.

Using whole cell antigens prepared from the established lines of human malignant fibrous histiocytoma (MFH), the authors have generated two different monoclonal antibodies (FU3 and FU4) by a mouse hybridoma technique. By indirect immunoperoxidase in frozen tissue sections, FU3 and FU4 revealed strong staining of perivascular mesenchymal cells and fibroblasts. In the spleen FU3 stained perivascular cells of the ellipsoids and the marginal zone of the lymph follicles. Macrophages in granulation tissues as well as monocytes and other blood cells in normal peripheral blood were uniformly negative for the antigen detected by FU3. Among various soft-tissue tumors, MFH and liposarcoma reacted strongly with FU3 and FU4, but synovial sarcoma revealed no reaction with either of the antibodies. Immunoelectron-microscopic studies demonstrated positive reactions with FU3 and FU4 on the surface of MFH cell membrane, which suggests that these antibodies recognized cell surface antigens. In conclusion, MFH shares antigenicity with perivascular mesenchymal cells and fibroblasts as well as liposarcoma. MFH and liposarcoma may have a common origin from the perivascular mesenchymal cells that are supposed to have a potential for multidirectional differentiation.

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Juvenile xanthogranuloma of the hand.

A case of juvenile xanthogranuloma in the hand is reported. The lesion was atypical clinically, being sited in the hand and the tumour was so large that it extended from the palmar to the dorsal surface. Curettage was performed at seven weeks after birth and histological examination established a diagnosis of juvenile xanthogranuloma. The post-operative course was uneventful, and the residual lesion had disappeared spontaneously four years after operation.

Female↗

Ontogeny of preproenkephalin mRNA expression in the rat retina.

Endogenous opioid systems (i.e. opioid peptides and opioid receptors) modulate developmental events in the neonatal mammalian retina. In the present study, the mRNA encoding preproenkephalin A (PPE), the prohormone for the opioid growth factor (OGF), [Met5]-enkephalin, was studied in the developing and the adult retinas of rats. Northern analysis indicated the presence of a 1.4-kb message in the developing and adult retinas corresponding to rat PPE mRNA. Quantitation showed that PPE message was present on postnatal day 1 at 5% of the adult level, and increased during development until the adult quantity was reached by postnatal day 27. In situ hybridization experiments first detected the presence of PPE mRNA in retinal tissues during late gestation. In late prenatal and neonatal retinas, PPE message was associated with areas of the developing retina containing proliferating neuroblasts and postmitotic cells. Later in development, message appeared to be located primarily within the inner retina, with abundant PPE mRNA associated with putative horizontal cells of the inner nuclear layer (INL). The adult retina showed a similar pattern of PPE gene expression in the cells of the INL. These findings document that the gene expression in the retina for PPE begins in the fetus, continues during retinal development, and coincides with the presence of a PPE mRNA derivative ([Met5]-enkephalin) that regulates DNA synthesis during retinal ontogeny. Our results are also the first to show the presence of PPE message in the adult mammalian retina, suggesting transcription of an opioid gene in the mature visual system.

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Ontogeny of the opioid growth factor, [Met5]-enkephalin, and its binding activity in the rat retina.

The endogenous opioid peptide [Met5]-enkephalin is a tonically active opioid growth factor (OGF) with an inhibitory action on DNA synthesis in the developing rat retina. In this study, the ontogeny of the spatial and temporal expression of OGF and its binding activity was examined. OGF-like immunoreactivity was detected in the retina at gestation day (E) 20, but not at E18, and was localized to ganglion cell and neuroblast layers; immunochemical reaction was no longer seen in the retina by postnatal day 6. Native OGF was further identified and characterized by high-performance liquid chromatography (HPLC) studies and immunodot assays, which revealed that [Met5]-enkephalin was present in the neonatal, but not adult, rat retina. OGF binding activity was detected as early as E18 using [125I]-[Met5]-enkephalin and in vitro receptor autoradiography. Little OGF binding activity was noted for prenatal retinas, but appreciable activity was observed from birth to postnatal day 4; no OGF binding could be detected after postnatal day 5 or in the adult. These results reveal the transient appearance of the OGF, [Met5]-enkephalin, and its receptor binding activity in the developing mammalian retina, and show that their ontogeny coincides with the timetable of DNA synthesis of retinal neuroblasts.

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