Brown adipose tissue activity dissipates energy as heat, and there is evidence that lack of the retinoblastoma protein (pRb) may favor the development of the brown adipocyte phenotype in adipose cells. In this work we assessed the impact of germ-line haploinsufficiency of the pRb gene (Rb) on the response to high fat diet feeding in mice. Rb^+/- mice had body weight and adiposity indistinguishable from that of wild-type (Rb^+/+) littermates when maintained on a standard diet, yet they gained less body weight and body fat after long-term high fat diet feeding, coupled to reduced feed efficiency and increased rectal temperature. Rb haploinsufficiency ameliorated insulin resistance and hepatosteatosis after high fat diet in male mice, in which these disturbances were more marked than in females. Compared to wild-type littermates Rb^+/- mice fed a high fat diet displayed higher expression of peroxisome proliferator-activated receptor (PPAR) as well as of genes involved in mitochondrial function, cAMP sensitivity, brown adipocyte determination and tissue vascularization in white adipose tissue depots. Furthermore, Rb^+/- mice exhibited signs of enhanced activation of brown adipose tissue and higher expression levels of PPAR in liver and of PPAR in skeletal muscle, suggestive of an increased capability for fatty acid oxidation in these tissues. These findings support a role for pRb in modulating whole body energy metabolism and the plasticity of the adipose tissues in vivo, and constitute first evidence that partial deficiency in the Rb gene protects against the development of obesity and associated metabolic disturbances.