Recent studies have demonstrated that hemichannels, which form gap junctions when paired from apposing cells, may serve additional roles when unpaired including cell adhesion and paracrine communication. Hemichannels in mammals are formed by connexins or pannexins, while in insects they are formed by pannexin homologues termed innexins. The formation of functional gap junctions by insect innexins has been established, although their ability to form functional nonjunctional hemichannels has not been reported. Here the characteristics of nonjunctional hemichannels were examined in three lepidopteran cell types, two cell lines (High Five and Sf9) and explanted hemocytes from Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae). Selective fluorescent dye uptake by hemichannels was observed in a significant minority of cells, using fluorescence microscopy and flow cytometry. Carbenoxelone, an inhibitor of mammalian junctions, disrupted dye uptake, while flufenamic acid and mefloquine did not. The presence of Ca(2+) and Mg(2+) in the media increased hemichannel activity. Additionally, lipopolysaccharide, a stimulator of immune activity in lepidopterans, decreased dye uptake. These results demonstrate for the first time the activity of nonjunctional hemichannels in insect cells, as well as pharmacological tools to manipulate them. These results will facilitate the further examination of the role of innexins and nonjunctional hemichannels in insect cell biology, including paracrine signaling, and comparative studies of mammalian pannexins and insect innexins.

The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), preferentially oviposits and feeds on ears of corn (Zea mays L.) and can be managed using transgenic hybrids that produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt). Concentrations of Bt proteins can vary spatially and temporally in plant tissues, creating a heterogeneous environment that can increase the risk of resistance development. We planted small-plot trials of nine Bt and non-Bt corn hybrids in South Carolina in 2016 and 2017 and investigated the development, survival, feeding injury, and feeding behavior in corn ear tissues. ELISA was used to quantify the concentrations of Cry1F and Cry2Ab2 in young silk, old silk, maternal tip tissue, kernels, and husk. Cry1F and Cry2Ab2 significantly varied with silk age and both proteins were generally highest in the silk and tip tissue. Hybrids with pyramided proteins significantly reduced feeding injury to the silk, tip, and kernel ear tissues, which was less apparent with single Bt protein hybrids. The pyramided hybrid expressing Vip3A incurred no injury to either the ear tip or kernels, and only eight 1^st instar larvae were collected in the silk of 520 sampled ears. Age of larvae significantly varied among ear tissues but not between hybrids. Depending on hybrid family, mean larval instar in the silk, tip, and kernels was 1^st or 2^nd, 3^rd, and 5^th, respectively. Instar-specific feeding penetrance into corn ears increased with age but did not differ between hybrids. We characterized the instar- and tissue-specific feeding behavior of H. zea larvae but did not detect differences in feeding behavior between Bt and non-Bt hybrids. Implications for resistance management strategies such as seed mixtures are discussed.