In this study, we investigated the influence of the length of nest deprivation period (3 vs. 6 d) on the ability to renest of incubating hens. We focused on the hens' behaviour, particularly on nesting and calls, on their prolactin, luteinizing hormone (LH) and oestradiol concentrations. Nest deprivation induced a drop of prolactin and an increase of LH plasmatic concentrations in hens. These changes in circulating pituitary hormones were followed by changes in ovarian function: the persistent rise of plasmatic oestradiol gave evidence of the resumption of ovarian activity. After nest deprivation, the number of clucks increased significantly and food calls appeared; these results demonstrate that ‘maternal’ calls can be emitted outside the maternal context. Our results suggest that the onset of typical maternal calling is strongly controlled by internal state such as plasmatic hormonal concentrations, independently of social stimulation. None of the 10 hens deprived for 6 d resumed incubation when given the opportunity, whereas, after a 3 d deprivation period, three out of 10 hens renested and two hens incubated sporadically and then gradually abandoned their nests. Long periods of nest deprivation appear to disrupt the habits of sitting and nesting. Before nest reopening, all the hens presented low levels of plasmatic prolactin. Plasma prolactin concentrations of the renesting hens increased after nest boxes were reopened and returned to levels found in incubating hens. We suggest that pituitary prolactin rather than plasmatic prolactin is responsible for the maintenance of incubating potential in hens deprived of their nests.