A possibility for differentiating dopamine from noradrenaline in tissue sections by microspectrofluorometry.
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Animals↗
Biomedical subjects
B Ehinger
Publications and source records attributed to B Ehinger.
Histochemical studies on the metabolism of L-DOPA and some related substances in the exocrine pancreas.
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5-Hydroxytryptophan↗
Function and structure in retinal transplants.
Embryonic mammalian donor retina transplanted into the subretinal space of a mature host develops into a graft with well-organized, but atypical retinal structure. We tested the effect of this organization on rabbit-to-rabbit graft functional properties, isolating the graft to avoid contamination of graft responses by host retinal activity. Transient ON or ON-OFF spike-like responses and local electroretinograms (L-ERGs) were recorded simultaneously via a single electrode on the graft surface. These response components depended on stimulus diameter, sometimes in a way indicating antagonistic center-surround receptive field organization and spatial tuning (43%). Other times, the responses were an increasing function of stimulus diameter which saturated for large spots (57%). Response amplitudes were a monotonically-increasing function of light intensity over the narrow range tested. The L-ERGs were reminiscent of the proximal negative response or M-wave seen in normal retinas, which reflect light-induced amacrine cell activity. Thus, for the first time, we have shown that these subretinal grafts possess light-transduction and complex functional properties like those in normal retinas. They also possess the cellular complement and synaptic microcircuitry needed to form these physiological properties, Therefore, these results demonstrate a functional ability and capacity in transplants that is required if nerve cell transplantation surgery is to be done with therapeutic aims.
Action Potentials↗
Ultrastructural circuitry in retinal cell transplants to rat retina.
The development of five transplants of fetal retinal tissue to adult rat eyes was examined with the electron microscope. The transplants were of 9 to 10 weeks total age after conception in four cases and 20 weeks in one case. They were at stage E15 when transplanted. Transplants developed in both the epiretinal and subretinal spaces. The transplants were heterogeneously developed with some parts showing almost normal differentiation and others little. Subretinal transplants examined in this study were more developed than epiretinal grafts. Photoreceptor cells developed both inner and outer segments. Their synaptic terminals possessed output ribbon synapses with postsynaptic processes similar to those seen in normal retinas. In regions corresponding to the inner plexiform layer, the adult complement of synapses was seen, including advanced features such as serial synapses as well as reciprocal synapses at bipolar cell dyads. Incompletely differentiated synapses of both the amacrine and bipolar cell types were often observed, especially in the rat epiretinal transplants. Ganglion cell processes could not be identified with certainty. Although transplant cells were adjacent to host photoreceptor cells and pigment epithelium, obvious specializations or interactions were not observed. The experiments suggest that embryonic rat retinal cell transplants develop most or perhaps all of the structural components and neuronal circuitry necessary to transduce light and process some visual information.
Animals↗
Full-thickness retinal transplants: a review.
Embryonic full-thickness rabbit neuroretinal sheets were transplanted to the subretinal space of adult hosts. This was accomplished by using a new transplantation technique involving vitrectomy and retinotomy. The grafts were followed from 10 to 306 days after surgery and were then examined by different histological techniques. In the light microscope, the transplants were seen to develop the normal retinal lamination and fusion with the host retina, especially after long survival times. Ultrastructurally, normal photoreceptor outer segments, well integrated with the host retinal pigment epithelium, were found. Growth cones were present in the zone of fusion between graft and host retina. Immunohistochemical labeling revealed many of the normal retinal components not previously found in retinal transplants, and graft-host connections between neurons in the rod pathway were seen. The morphology of vibratome-sectioned neuroretinal sheets as well as adult full-thickness grafts was also examined. These transplantation types showed less of the normal morphology compared with embryonic full-thickness grafts. The immunogenicity of embryonic full-thickness and fragmented grafts was compared using major histocompatibility complex immunolabeling. Fragmented grafts elicited a response from the host immune system similar to a chronic transplant rejection. This reaction was absent in the full-thickness grafts which is in accordance with their good long-term survival.
Animals↗
Ultrastructure of retinal cells transplanted to the rabbit choroid.
PURPOSE: Allogenic rabbit-to-rabbit retinal cell transplants survive in the choroid, which is not as expected because it has not been shown that this is an immune-privileged site. We have therefore examined the ultrastructure of such transplants, looking for features that might explain the phenomenon. METHODS: Rabbit retinal tissue fragment transplants were produced with previously described methods. The donor age was 15 days and the transplants were examined by standard electron microscopy when the transplants were 1-2 months (3 transplants) or 3-4 months old, of postconception age (3 transplants). RESULTS: The transplants survived and developed as expected from previous observations. Rosettes were seen, but they were not as common as in transplants produced with the same technique in the subretinal space of rabbits. Photoreceptor outer segments were not seen in the transplants. At 1 month, there was an incomplete sheath of Müller cells around the transplants, and a complete one at 3-4 months. There was also a well-developed basement membrane around the transplant at 3-4 months, but less so at 1 month. Blood vessels did not enter the transplant. The fenestrations in the choriocapillaris were not affected as long as the pigment epithelium was normal. CONCLUSIONS: The enclosure of the transplants by Müller cells might help to insulate them from the immune system of the host, but it is a late phenomenon and it is not likely to have much effect for the first few weeks after the transplantation. We suspect that either the rabbit choroid is an immune-privileged site, even though there is no previous direct evidence for this, or that the retinal tissue itself is responsible for the prolonged survival at this site.
Animals↗
Expression of nitric oxide synthase during the development of RCS rat retinas.
Nitric oxide (NO) has been reported to be both neurodestructive and neuroprotective in the central nervous system and could possibly play an important role in neurodegenerative disorders. On the assumption that NO synthesis may influence degenerative processes in the retina, we have examined the development and distribution of nitric-oxide-synthase(NOS)-immunoreactive cells in developing Royal College of Surgeons (RCS) rat retinas, which is an animal model for retinal degeneration. An antibody against constitutive neuronal NOS was used for immunocytochemistry on RCS rat retinas from postnatal (PN) days 3, 7, 10, 14, 35, 70 and 281 and compared with that in the normal rats of PN days 3, 7, 10, 14, 54 and adults. Immunoreactive cells were not seen in PN 3 retinas but were distinctly seen in the PN 7 retina along with a plexus in the inner plexiform layer. In both groups (normal and RCS rats) a distinct sublayering of the plexus in the inner plexiform layer could be seen at PN 10, which became more distinct at PN 14. The immunoreactive cells were detected also in the oldest retina examined, which was PN 281 in the case of RCS rats. In both groups, certain amacrine cells, certain bipolar cells and certain horizontal cells were found to be immunoreactive. In conclusion, the developmental timetable of the NOS immunoreactivity was identical in the normal and the RCS rat retinas. The NOS-immunoreactive cells persisted in the RCS retinas even when the retina had degenerated extensively. Abnormalities with the inducible isoforms of NOS cannot be ruled out from this study. We conclude that the chronological and qualitative development of the constitutive neuronal NOS immunoreactivity is normal in RCS rat retinas.
Animals↗
Cellular organization in retinal transplants using cell suspensions or fragments of embryonic retinal tissue.
We have investigated the cellular organization in two different types of retinal transplants using cell type-specific monoclonal antibodies. Both fragments and cell suspensions of E17-E19 Sprague-Dawley rat retina were transplanted to a subretinal site in congenic adult rat hosts. After a survival time of 28 days, the transplants were stained by immunocytochemistry with antibodies against rhodopsin, which stained rods; with antibodies against HPC-1, which stained amacrine cells and outer and inner plexiform layers; and with antibodies against vimentin, which stained Müller cell fibers and horizontal cells. In the host retina, the distribution of immunocytochemical staining was similar, irrespective of transplantation technique. In the transplants, the anti-rhodopsin staining showed that fragment transplants developed photoreceptors in rosettes, whereas in cell suspension transplants, this staining showed a scattered distribution of photoreceptors. The HPC-1 staining showed that regions corresponding to the inner nuclear layer surrounded both types of transplants and made large invaginations into them. In one case, using the cell suspension technique, fibres were found to run from the inner plexiform layer of the transplant to the outer plexiform layer of the host. The vimentin staining revealed a disorganized array of Müller cell fibres in both types of transplants, but with some concentration to the regions corresponding to the inner plexiform layer.
Animals↗
Cell proliferation in retinal transplants.
The MIB-1 antibody against a nuclear protein Ki-67 was used to study the proliferation of cells in the rabbit retinal transplants. Fragmented pieces of embryonic day 15 rabbit retinas were transplanted into the subretinal space of adult rabbits and allowed to survive for different times. Fragmented donor tissue starts organizing in rosettes 1 day after transplantation. The transplanted cells continue to proliferate in the host eye and their pattern of proliferation resembles that of normal developing retina, suggesting that the factors responsible for the proliferation pattern are preserved after transplantation. The dividing cells in metaphase line up in the luminal layers of the rosettes. Certain cells become postmitotic in the regions corresponding to the inner retina first, followed by the cells in the luminal layers of rosettes. Cells in the regions between the rosettes, corresponding to the inner nuclear layer, presumably the Müller cells, proliferate significantly for the equivalent age of postnatal day 2. Few cells in these regions proliferate for at least the equivalent age of postnatal day 11 in transplants. There is a layer of nonproliferating, degenerating cells in the transplant situated close to the host retina. However, some cells in this layer, situated at the host-graft interface, proliferate. These cells proliferate for a long time possibly indicating gliosis.
Animals↗
Corticosteroid suppression of trauma-induced hyaluronan in rabbit cornea and aqueous.
BACKGROUND: Corneal and aqueous hyaluronan have recently been shown to react in response to several different types of trauma, including cataract surgery. In order to find ways to influence the reaction, we have evaluated the effect of topical dexamethasone (Isopto-Maxidex, Alcon Universal Ltd, Fort Worth, Tex) or indomethacin (Confortid, Dumex Ltd, Copenhagen, Denmark) on the postoperative hyaluronan concentration in rabbit cornea and aqueous after extracapsular lens extraction. METHODS: The drugs were administered as topical eye drops three times daily. The hyaluronan concentration in rabbit cornea and aqueous after extracapsular lens extraction was measured with a radioligand assay. RESULTS: Dexamethasone treatment (1 mg/mL) significantly suppressed the increase in corneal hyaluronan seen after extracapsular lens extraction, 2 (p < or = 0.0022) and 3 weeks (p < or = 0.0002) after surgery, while indomethacin did not induce any significant difference at 2 weeks. When the dexamethasone concentration was lowered to 0.1 and 0.2 mg/mL, there was still a significant decrease (p < or = 0.009) in hyaluronan concentration, but at lower concentrations of dexamethasone (0.01 and 0.02 mg/mL), no significant decrease was seen. The increase in aqueous hyaluronan concentration seen 2 days after surgery in untreated eyes was significantly lowered by both dexamethasone (p < or = 0.0076) and indomethacin (p < 0.036). CONCLUSIONS: Dexamethasone lowers reactive corneal and aqueous hyaluronan concentration in vivo after extracapsular lens extraction.
Administration, Topical↗
[Chronic-idiopathic enteropathy in cats--a case control study].
Aim of this study is to clinically, microbiologically and histopathologically characterize inflammatory bowel disease (IBD) in the cat. Nine cats with chronic persistent or intermitent vomitus and diarrhea were examined between 1998 and 2001. All cats had a thuorough diagnostic workup performed. Full thickness biopsies from stomach, duodenum, jejunum, ileum and colon were surgically obtained for histopathological examination. Duodenal juice was obtained by direct aspiration for microbiologic qualitative and semiquantitative examination. Seven cats euthanized for other medical reasons were used as controls. Six cats had a lymphoplasmacytic IBD and three an eosinophilic IBD. Four cats with IBD had additional diseases diagnosed. Three cats with IBD had elevated bacterial counts. Retrospectively no correlation could be found between clinical symptoms and histopathological results. Serum TLI was not able to differentiate chronic pancreatitis and IBD. Serum folic acid and cobalamin did not correlate with the distribution of lesions in the gastrointestinal tract. Finally, no correlation was found between bacterial counts in the proximal duodenum and IBD.
Animals↗
Influence of radial keratotomy on endogenous hyaluronan in cornea and aqueous humour.
BACKGROUND: Hyaluronan is known to appear in corneal wound tissue and is probably involved in the healing process. We measured the changes of endogenous hyaluronan in rabbit cornea and aqueous after radial keratotomy. METHODS: Corneal and aqueous hyaluronan concentrations were measured at different time intervals after radial keratotomies. Hyaluronan was extracted from corneal tissue with two methods employing either pronase or NaCl and measured with a radioimmunoassay. Histochemical staining for hyaluronan was done on the operated eyes. RESULTS: Changes in corneal hyaluronan showed a similar time course with either extraction method. Maximal concentration was reached after 7 to 14 days with a return to normal values in 60 days. Aqueous hyaluronan and proteins were not significantly altered after surgery. Histochemical staining of the corneas demonstrated hyaluronan around the incisions along their entire extent. Blood vessels were absent in the wounds. An increase in elongated fibroblast-like cells was seen in the wound area. CONCLUSIONS: Both quantitatively and histochemically, there is a pronounced increase of corneal hyaluronan in avascular, radial keratomy wounds in the rabbit. The findings suggest a role of hyaluronan in corneal wound healing.
Animals↗