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Retinal pigment epithelial (RPE) cells are among the most actively phagocytic cells in nature. Primary RPE and stable RPE cell lines provide experimental model systems that possess the same phagocytic machinery as RPE in situ. Upon experimental challenge with isolated photoreceptor ...

The identification of stem/progenitor cells within the retinal neural environment has opened up the possibility of therapy via cellular replacement and/or reprogramming of resident cell populations (1–4). Within the neuro-retinal niche, following injury or in disease states (i ...

Calcium acts as a prominent second messenger in virtually every cell type and modulates a plethora of cell functions. Thus, Ca2+ microfluorimetry became a valuable tool to assess information about mechanisms involved in the regulation of the intracellular calcium level in research on li ...

Ca2+ is an important regulator of many cell functions including proliferation, apoptosis, movements, secretion, contraction, excitation, and differentiation. The regulation of these different cell functions is encoded by the specific temporal and spatial distribution of Ca2+ ...

Combinatorial binding of transcription factors (TFs) and cofactors to specific regulatory regions of target genes in vivo is an important mechanism of transcriptional regulation. Chromatin immunoprecipitation (ChIP) is a powerful technique to detect protein binding to spec ...

Transcription factors control gene expression by binding to noncoding regions of DNA known as �cis-regulatory elements (CREs; i.e., enhancer/promoters). Traditionally, cis-regulatory analysis has been carried out via mouse transgenesis which is time-consuming and nonquan ...

Subretinal injections in mice become increasingly important. Currently, the most prominent application is in gene therapy of inherited eye diseases by means of viral vector delivery to photoreceptors or the retinal pigment epithelium (RPE). Since there are no large animal models for mo ...

Adeno-associated virus (AAV) vectors are in wide use for in vivo gene transfer for the treatment of inherited retinal disease. AAV vectors have been tested in many animal models and have demonstrated efficacy with low toxicity. In this chapter we describe some of the recent methods for small-sca ...

The inner blood–retina barrier (iBRB) is essential in restricting the movement of systemic components such as enzymes, anaphylatoxins, or pathogens that could otherwise enter the neural retina and cause extensive damage. The barrier has evolved to confer protection to the delicate mi ...

Genetic lineage tracing is an invaluable tool to demonstrate and measure neogenesis of beta cells from putative precursor cells. Cre-Lox recombination technology can be used for indelible labeling of a cohort of cells and following the fate of these cells and their progeny in animal models. H ...

Restoring a functional β cell mass in diabetes patients by β cell transplantation or stimulation of β cell regeneration are promising approaches. It requires knowledge on the mechanisms of β cell neogenesis, an issue that is still quite controversial. Postnatal islet regeneration may or m ...

Retinal digestion is a commonly used method for studying experimental diabetic retinopathy in animal models. The method allows to assess qualitatively and quantitatively the morphology of the retinal vasculature, including characteristics of endothelial cells and pericyt ...

Several mouse strains are diabetic already at the juvenile age or develop diabetes mellitus during their life. Before these strains become diabetic, they often show several or all features of the metabolic syndrome, which is very similar to the etiology of diabetes in humans. Under the assumpt ...

Beta cell destruction in autoimmune diabetes is accompanied by the presence of autoantibodies and autoreactive T cells against beta cell antigens. Autoantibodies to insulin are predictive of future diabetes in man and in the non-obese diabetic mouse model. Furthermore, the detection ...

Glucose disposal in skeletal muscle is a major target for insulin action and assessment of insulin-regulated glucose uptake under in vitro conditions allows the direct determination of insulin sensitivity in this organ. For this purpose, a variety of muscle preparations from differe ...

A tracer technique referred to as “pancreatic-blood glucose clamp” allows assessment in response to a change in blood glucose, insulin, and/or glucagon of whole body glucose disposal, endogenous glucose production, specific tissue/organ glucose uptake and storage, and insulin sec ...

The feasibility of investigating glucose tolerance and insulin action and secretion in vivo in mouse models has provided major insights into both type 2 diabetes pathogenesis and the identification of novel strategies to treat this common disorder. When initial studies provide evid ...

For the regulation of beta-cell function ion channels are of outstanding importance. Beta cells are specialized to convert changes in blood glucose concentration to an adequate secretory response. To achieve this, nutrient-induced alterations of electrical activity are direct ...

The kinetics of insulin secretion, not just the total amount, is of decisive relevance for the physiological regulation of glucose homeostasis. Thus to characterize the relevant features of the secretory response to an insulinotropic stimulus a method is needed which is able to resolve the ...

Nonhuman primates provide the ideal animal model for discovering and examining further the mechanisms underlying human type 2 diabetes mellitus. In all aspects studied to date the nonhuman primate has been shown to develop the same disease with the same features that develop in overweight ...