Virus injections were performed as described previously (19) . Briefly, mice were anaesthetized with isoflurane using stereotaxic instrument (David Kopf instruments, Tujunga, CA, USA) and scalp was incised carefully before drilling the skull for injection. Two hundred and fifty to three hundred nanoliters of intracranial injection were performed with a pulled glass pipette (Drummond Scientific, Wiretrol, Broomall, PA, USA) having 40-50 lm tip diameter. Recombinant adeno-associated virus production was conducted as described previously (33) . Cre-dependent AAV vectors rAAV2/1-EF1a-DIO-hM3D(Gq)-mCherry and rAAV2/8-FLEX-GFP were purchased from Addgene http://www.addgene.org/. rAAV2/1-EF1a-DIO-hM3D(Gq)-mCherry (2 Â 10 12 genomic copies/ml) or rAAV2/8-FLEX-GFP (10 14 genomic copies/ml) was injected at coordinates around the ARC (bregma: À1.25 mm, midline: 60.25 mm, dorsal surface À5.20 and À5.70 mm). Injection speed was set to 30 nl/min by a micromanipulator (Narishige, East Meadow, NY, USA). Following first intracranial injection at À5.70 mm dorsal surface, another intracranial injection was performed at À5.20 mm dorsal surface, allowing 10 min time for each injection. Injection pipette was slowly withdrawn and the scalp was stitched. Animals were allowed 14 days for recovery and transgene expression.
Oncul M., Dilsiz P., Ates Oz E., Ates T., Aklan I., Celik E., Sayar Atasoy N, & Atasoy D. (2018). Impaired melanocortin pathway function in Prader-Willi syndrome gene-Magel2 deficient mice. Human molecular genetics, 27(18).
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