These findings provide evidence that in murine ophthalmic arteries the muscarinic M3 receptor subtype mediates cholinergic endothelium-dependent vasodilation and endothelium-independent vasoconstriction.

We tested the specificity of three commercially available antibodies (AB) against individual α1-adrenergic receptor subtypes (α1-ARST). We used these ABs to localize the α1-ARST proteins by immunohistochemistry in paraffin-embedded murine tissues. The specificity of the ABs was tested by comparing staining patterns in tissues from wild-type mice with those in corresponding tissues from mice with gene-targeted disruption of the respective α1-ARST, one of the most rigorous negative controls. None of the tested ABs proved to be specific for the indicated target antigen. We conclude that the tested ABs are unsuitable for immunohistochemistry in paraffin-embedded murine tissues.

BACKGROUND AND PURPOSEThe α1-adrenoceptor family plays a critical role in regulating ocular perfusion by mediating responses to catecholamines. The purpose of the present study was to determine the contribution of individual α1-adrenoceptor subtypes to adrenergic vasoconstriction of retinal arterioles using gene-targeted mice deficient in one of the three adrenoceptor subtypes (α1A-AR −/− , α1B-AR −/− and α1D-AR −/− respectively). EXPERIMENTAL APPROACHUsing real-time PCR, mRNA expression for individual α1-adrenoceptor subtypes was determined in murine retinal arterioles. To assess the functional relevance of the three α1-adrenoceptor subtypes for mediating vascular responses, retinal vascular preparations from wild-type mice and mice deficient in individual α1-adrenoceptor subtypes were studied in vitro using video microscopy. KEY RESULTSRetinal arterioles expressed mRNA for all three α1-adrenoceptor subtypes. In functional studies, arterioles from wild-type mice with intact endothelium responded only negligibly to the α1-adrenoceptor agonist phenylephrine. In endothelium-damaged arterioles from wild-type mice, phenylephrine evoked concentration-dependent constriction that was attenuated by the α1-adrenoceptor blocker prazosin. Strikingly, phenylephrine only minimally constricted endothelium-damaged retinal arterioles from α1B-AR −/− mice, whereas arterioles from α1A-AR −/− and α1D-AR −/− mice constricted similarly to arterioles from wild-type mice. Constriction to U46619 was similar in endothelium-damaged retinal arterioles from all four mouse genotypes. CONCLUSIONS AND IMPLICATIONSThe present study is the first to demonstrate that α1-adrenoceptor-mediated vasoconstriction in murine retinal arterioles is buffered by the endothelium. When the endothelium is damaged, a vasoconstricting role of the α1B-adrenoceptor subtype is unveiled. Hence, the α1B-adrenoceptor may represent a target to selectively modulate retinal blood flow in ocular diseases associated with endothelial dysfunction. Abbreviations α1A-AR−/− mouse, α1A-adrenoceptor knockout mouse; α1B-AR −/− mouse, α1B-adrenoceptor knockout mouse; α1D-AR −/− mouse, α1D-adrenoceptor knockout mouse; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; U46619, 9,11-dideoxy-9α,11α-methanoepoxy PGF2α

The retina is a part of the central nervous system, a thin multilayer with neuronal lamination, responsible for detecting, preprocessing, and sending visual information to the brain. Many retinal diseases are characterized by hemodynamic perturbations and neurodegeneration leading to vision loss and reduced quality of life. Since catecholamines and respective bindings sites have been characterized in the retina, we systematically reviewed the literature with regard to retinal expression, distribution and function of alpha1 (α1)-, alpha2 (α2)-, and beta (β)-adrenoceptors (ARs). Moreover, we discuss the role of the individual adrenoceptors as targets for the treatment of retinal diseases.