Discriminating between conspecific and heterospecific communication signals has important implications for evolution. The benefits of such discriminations are clear for sympatric congeners, but if conspecific display recognition (CDR) has a cost, it should be relaxed in species that have evolved in isolation. Of the nine lava lizard species (Microlophus spp.) endemic to the Galápagos Islands, all are thought to have evolved in allopatry and none overlap in geographic distribution. Although prior research failed to reveal CDR in male M. grayii on Floreana Island, male M. indefatigabilis on Santa Cruz Island showed a response bias toward conspecific displays under the same experimental conditions. In the present study we tested for CDR in two additional species: M. albemarlensis on Isabella Island and M. bivittatus on San Cristóbal Island. Subjects were presented with computer-controlled robots that either performed the conspecific display, a reversed-inverted conspecific display, or a display from an Anolis lizard species. Results revealed evidence of CDR in male M. albemarlensis, but no such evidence in male M. bivittatus. Interestingly, bathymetric data provide a potential explanation for CDR in M. indefatigabilis and M. albemarlensis: a land bridge between Santa Cruz and Isabella might have resulted in secondary contact and reinforcement in these two species during Pleistocene glacial maxima. Another possibility is that intraspecific selection for discrimination among males has caused heterospecific displays to be viewed as low-fidelity conspecific displays. Nevertheless, our findings are consistent with CDR being lost early in Galápagos Microlophus evolution, as predicted for allopatric speciation, and re-emerging later only where selection favored the discrimination of male displays.