Gap junction proteins are membrane channels that connect the cytoplasm of neighboring cells, allowing for cellular communication through the selective transfer of small molecules. Gap junctions coordinate an array of multicellular processes, including tissue patterning and potentially cellular immune responses. We have investigated the role of gap junctions in hemocytes of the lepidopteran Pseudoplusia includens. Hemocytes are involved in the primary lepidopteran immune response to multicellular invaders. After recognizing an invader, layers of hemocytes surround and subsequently kill the invader. Successful encapsulation of invaders requires coordination of the hemocytes. Insect gap junctions are encoded by innexin genes. We have isolated two novel innexins from P. includens. Low degeneracy PCR primers were designed based on highly conserved sequence of Drosophila innexins and used to isolate partial transcripts. Based on sequence analysis, these partial transcripts were determined to be innexin 1 (pi-inx1) and innexin 2 (pi-inx2), orthologues to innexins from other insects. Learning more about these highly conserved sequences may help elucidate developmental patterns in insects, as well as provide useful models for the study of gap junctions in insect immunity. Analysis of innexin transcripts suggests that levels vary between tissues, with higher levels of pi-inx2 transcription occurring in hemocytes than in other tissues analyzed. In addition, pi-inx2 relative transcription levels increase in hemocytes following immune challenge by various pathogenic agents. Taken together, these data support a role for gap junctions in the coordination of immune responses.