Monoamine brain content of mainland and island populations of Podarcis siculus

In accordance with the permit of the Ministry of Environment and Energy (Class: AP /L612-07 I 18-481 65; No: 517-07-l-l-l-18-4), and the permit of the Ethical committee of the Faculty of Science, University of Zagreb, Croatia (No: 251-58-10617-18-16), animals (10 males from each location) were captured by noosing, and transported to the laboratory in cloth bags. Since the population on islet Pijavica is strictly protected, this was the maximum number of animals we were permitted to collect. In the laboratory, animals were maintained in individual terraria (40x30cm) with peat as a substrate, fed every two days with crickets (Acheta domestica), that were supplemented with calcium and vitamins, with water available ad libitum. The light cycle was the same as the natural cycle (14:10 h, light: dark). Room temperature varied between 22°C during the night and 30° C during the day, to simulate natural day-night temperature variation. Lizards were allowed to habituate to laboratory conditions for one week before being subjected to behavioral testing. Brain samples were collected two weeks after the last behavioral test (beginning of July 2018). The experiments were conducted in accordance with the Directive of the European Parliament and of the Council (2010/63/EU) and the Croatian Animal Protection Law (“Narodne Novine”, 102/17 and 32/19). All efforts were made to reduce the number of animals used and to ensure animal welfare.