Adult and embryonic rabbit retinal sheets were transplanted into the subretinal space of adult rabbits. The transplants were either full-thickness with intact layering, or gelatin embedded and vibratome sectioned with the inner retina removed. The full-thickness grafts were positioned subretinally by means of a glass capillary in which they were partially folded. The vibratome sectioned ones were placed using a plastic injector in which the gelatin embedded graft was flat. The embryonic full-thickness grafts were followed clinically up to 3 months, and the other 3 transplant types up to 1 month postoperatively, after which the retina was sectioned and stained for light microscopy. Surgical complications were more common in eyes receiving vibratome sectioned grafts with 10 out of 34 eyes displaying blood in the vitreous. Four of these eyes also developed total retinal detachment. Out of 17 eyes receiving full-thickness grafts, only one displayed these complications. Histologically, 11 out of 13 embryonic full-thickness transplants revealed straight, laminated transplants with correct polarity, and with all normal retinal layers present. In these transplants, fusion with the host increased in time. Of the adult full-thickness transplants, only 1 out of 4 survived, and this graft showed signs of degeneration. The vibratome sectioned adult transplants in a few cases survived the first two postoperative weeks. In these grafts, both inner and outer retina were present, indicating an incomplete vibratome sectioning. With longer postoperative times, the number of surviving transplants in this group diminished considerably. All vibratome sectioned embryonic transplants developed into rosettes and sometimes also into laminated sections with reversed polarity. It can be concluded that in rabbits, the surgical technique used for vibratome sectioned transplants requires a larger sclerotomy and retinotomy, since they have to be kept flat in the transplanting instrument due to the surrounding gelatin. This technique is associated with a higher frequency of complications than the one used for full-thickness grafts which are more flexible and can be transplanted with a smaller instrument. Vibratome sectioning of embryonic grafts results in abnormal morphology and their adult counterparts only survive if the sectioning is incomplete. Adult full-thickness grafts show poor survival. Embryonic full-thickness transplants in the majority of cases develop into laminated retinas with layers parallel to the host retinal pigment epithelium. They also survive and integrate well with the host retina.
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http://dx.doi.org/10.1006/exer.1998.0582 | DOI Listing |
HardwareX
December 2024
Molecular and Cellular Biology Graduate Program, University of Massachusetts Amherst, Amherst, MA 01003, United States.
Slicing tissue samples into thin pieces is commonly used in histology analysis and more recently for organotypic culture when tissue samples are sliced alive. Currently available devices for slicing tissue samples are either designed for fixed tissue samples at low cryogenic temperatures (, Cryostats), or bulky and expensive (, vibratome), preventing them from routine lab usage. Here we report a cost-effective device designed to section live tissues for subsequent culture.
View Article and Find Full Text PDFCell Rep Methods
December 2024
Department of Biology, Georgetown University, Washington, DC 20057, USA. Electronic address:
Protein retention expansion microscopy (ExM) retains fluorescent signals in fixed tissue and isotropically expands the tissue to allow nanoscale (<70 nm) resolution on diffraction-limited confocal microscopes. Despite the numerous advantages of ExM, the protocol is time-consuming. Here, we adapted an ExM protocol to vibratome-sectioned brain tissue of Xenopus laevis tadpoles and implemented a microwave (M/W)-assisted protocol (ExM) to reduce the workflow from days to hours.
View Article and Find Full Text PDFMethods Mol Biol
October 2024
Bruker Daltonics GmbH & Co. KG, Bremen, Germany.
Mass spectrometry imaging (MSI) allows for label-free spatial molecular interrogation of tissues. With advances in the field over recent years, the spatial resolution at which MSI data can be recorded has reached the single-cell level. This makes MSI complementary to other single-cell omics technologies.
View Article and Find Full Text PDFCancer Med
September 2024
Department of Biomedical Sciences, Graduate School of Health Sciences, Hirosaki University, Hirosaki, Japan.
Background: Cancer initiation has long been "unknowable" in biology and medicine. In 1987, however, Moore and our research group observed single hepatocytes and minifoci that were strongly positive for glutathione S-transferase P-form (GST-P) in the rat liver as early as 2 to 3 days after initiation by diethylnitrosamine prior to the induction of GST-P foci and nodules. The induction of GST-P single hepatocytes, precursors of GST-P foci and nodules, was considered genetic.
View Article and Find Full Text PDFCold Spring Harb Protoc
August 2024
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) OT Gatersleben, Seeland 06466, Germany
Root anatomy plays a critical structural and functional role in the maize root system, and regulates edaphic stress tolerance. The function and genetic basis of several maize root anatomical traits for stress tolerance have been demonstrated. Leveraging root anatomical traits in maize thus holds great potential for developing cultivars with greater nutrient and water efficiency.
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