Glucagon-related peptides in the mouse retina and the effects of deprivation of form vision.
BACKGROUND
In chickens, retinal glucagon amacrine cells play an important role in emmetropization, since they express the transcription factor ZENK (also known as NGFI-A, zif268, tis8, cef5, Krox24) in correlation with the sign of imposed image defocus. Pharmacological studies have shown that glucagon can act as a stop signal for axial eye growth, making it a promising target for pharmacological intervention of myopia. Unfortunately, in mammalian retina, glucagon itself has not yet been detected by immunohistochemical staining. To learn more about its possible role in emmetropization in mammals, we studied the expression of different members of the glucagon hormone family in mouse retina, and whether their abundance is regulated by visual experience.


METHODS
Black wildtype C57BL/6 mice, raised under a 12/12 h light/dark cycle, were studied at postnatal ages between P29 and P40. Frosted hemispherical thin plastic shells (diffusers) were placed in front of the right eyes to impose visual conditions that are known to induce myopia. The left eyes remained uncovered and served as controls. Transversal retinal cryostat sections were single- or double-labeled by indirect immunofluorescence for early growth response protein 1 (Egr-1, the mammalian ortholog of ZENK), glucagon, glucagon-like peptide-2 (GLP-2), glucose-dependent insulinotropic polypeptide (GIP), peptide histidine isoleucine (PHI), growth hormone-releasing hormone (GHRH), pituitary adenylate cyclase-activating polypeptide (PACAP), secretin, and vasoactive intestinal polypeptide (VIP). In total, retinas of 45 mice were studied, 28 treated with diffusers, and 17 serving as controls.


RESULTS
Glucagon itself was not detected in mouse retina. VIP, PHI, PACAP and GIP were localized. VIP was co-localized with PHI and Egr-1, which itself was strongly regulated by retinal illumination. Diffusers, applied for various durations (1, 2, 6, and 24 h) had no effect on the expression of VIP, PHI, PACAP, and GIP, at least at the protein level. Similarly, even if the analysis was confined to cells that also expressed Egr-1, no difference was found between VIP expression in eyes with diffusers and in eyes with normal vision.


CONCLUSIONS
Several members of the glucagon super family are expressed in mouse retina (although not glucagon itself), but their expression pattern does not seem to be regulated by visual experience.
Mathis
Ute
U
Section of Neurobiology of the Eye, University Eye Hospital Tübingen, Calwerstrasse 7/1, 72076 Tübingen, Germany.
Schaeffel
Frank
F
D000818:Animals; D004195:Disease Models, Animal; D051766:Early Growth Response Protein 1 / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005749:Gastric Inhibitory Polypeptide / Q000378:metabolism; D005934:Glucagon / Q000378:metabolism; D004763:Glucagon-Like Peptides / Q000378:metabolism*; D013007:Growth Hormone-Releasing Hormone / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D009216:Myopia / Q000378:metabolism*; D010451:Peptide PHI / Q000378:metabolism; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D012683:Sensory Deprivation; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
C495402:Adcyap1 protein, mouse; D051766:Early Growth Response Protein 1; C486109:Egr1 protein, mouse; D010451:Peptide PHI; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D014660:Vasoactive Intestinal Peptide; D005749:Gastric Inhibitory Polypeptide; D004763:Glucagon-Like Peptides; D005934:Glucagon; D013007:Growth Hormone-Releasing Hormone