Effects of positive and negative lens treatment on retinal and choroidal glucagon and glucagon receptor mRNA levels in the chicken.
OBJECTIVE
It has been found in the chicken that the amount of retinal glucagon mRNA increases during treatment with positive lenses. Pharmacological studies support the idea that glucagon may act as a stop signal for visually induced eye growth. To gain more insight into the functional role of glucagon, the changes of glucagon and glucagon receptor mRNA concentrations in retina and choroid over time were studied. Furthermore, the abundance of glucagon and the glucagon receptor was studied in different fundal layers (retina, retinal pigment epithelium[RPE], choroid) and the blood.


METHODS
Semiquantitative real-time RT-PCR was used to measure glucagon and glucagon receptor mRNA levels in retina and choroid after positive and negative lens treatment for 2, 6, or 24 hours, by unilateral -7- or +7-D lenses. Contralateral eyes served as the control, and completely untreated animals provided further reference data. Intravitreal colchicine injections (which are known to reduce the number of glucagon cells sharply) were used to verify that the related decline in glucagon mRNA could be measured by real-time RT-PCR.


RESULTS
In the retina, treatment with -7-D lenses induced an initial upregulation of glucagon mRNA in both eyes, followed by a significant downregulation. The treatment with +7-D lenses showed a significant but transient downregulation in the control eye superimposed on a trend toward upregulation in the treated eye. However, the changes in glucagon mRNA expression were not confined to the lens-treated eyes but were also found, although sometimes to a lesser extent, in the non-lens-covered fellow eyes. There was evidence of a transient increase in glucagon receptor mRNA levels in lens-treated eyes after either -7- or +7-D lens wear. In the choroid, no effect of imposed defocus was detected. The injection of colchicine led to the destruction of approximately 75% of the glucagon amacrine cells but the mRNA level of retinal glucagon decreased by only approximately 50%. Glucagon receptor expression was found to be higher in the RPE than the retina and choroid whereas, in the blood, glucagon and glucagon receptor mRNA expression was below detection level.


CONCLUSIONS
The observed bidirectional regulation of glucagon mRNA in correlation with the sign of imposed defocus supports the idea that glucagon may act as a stop-and-go signal for eye growth. This is in line with a previous proposal based on studies of changes of the glucagon peptide content.
Buck
Christine
C
Section of Neurobiology of the Eye, University Eye Hospital, Tübingen, Tübingen, Germany.
Schaeffel
Frank
F
Simon
Perikles
P
Feldkaemper
Marita
M
D000818:Animals; D002645:Chickens; D002829:Choroid / Q000187:drug effects / Q000378:metabolism*; D003078:Colchicine / Q000633:toxicity; D015870:Gene Expression; D005934:Glucagon / Q000235:genetics* / Q000378:metabolism; D008297:Male; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000378:metabolism*; D018027:Receptors, Glucagon / Q000235:genetics* / Q000378:metabolism; D012030:Refractive Errors / Q000378:metabolism*; D012160:Retina / Q000187:drug effects / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012683:Sensory Deprivation
D012333:RNA, Messenger; D018027:Receptors, Glucagon; D012313:RNA; D005934:Glucagon; D003078:Colchicine