Identification of PLCβ and PKC in pheromone receptor neurons of Antheraea polyphemus

Abstract

Two proteins of the IP3 transduction pathway were identified by Western blots in homogenates of isolated pheromone-sensitive sensilla of the silkmoth Antheraea polyphemus. A 110 kDa protein was recognized by an antiserum raised against the Drosophila phospholipase C beta (PLC beta p121) and a 80kDa protein was labelled by an antiserum against a synthetic peptide of a conserved region of protein kinase C (PKC). Incubation of homogenized sensory hairs with the main sex pheromone component, (E,Z) 6-11 hexadecadienyl acetate, resulted in a 6-fold increase in the activity of PKC compared to controls without pheromone. In contrast, incubation with pheromone did not affect the activity of protein kinase A (PKA). Activation of PKC by the membrane permeable dioctanoylglycerol led to excitation of the pheromone-sensitive receptor neurons. These data support the current concept that pheromone perception of moths is mediated by the IP3 transduction pathway.

Intracellular regulation of the insect chemoreceptor complex impacts odor localization in flying insects

“…Obviously, a similar role for intracellular signalling in ORs of other pterygote orders must be demonstrated to give credence to this hypothesis. However, the aforementioned homology of Orco across several insect orders, and the expression of similar intracellular signalling proteins in the dendrite of other insects (Laue et al, 1997;Miura et al, 2005;Maida et al, 2000), suggests that such signalling may be common across the Pterygota.…”

“…PKC activity analyses were performed according to Ziegelberger et al (1990) and Maida et al (2000). Briefly, olfactory antennae of Drosophila were cut under a binocular light microscope using fine forceps and the isolated antennae were immediately stimulated with a headspace mixture of odours (ethyl hexanoate, ethyl butyrate, ethyl acetate, methyl acetate, 2,3-butanedione, 2-heptanone, 1-hexanol, pentyl acetate, ethyl-3-hydroxybutyrate).…”

High-Resolution Linkage Analyses to Identify Genes That Influence Varroa Sensitive Hygiene Behavior in Honey Bees

“…norpA encodes a phospholipase C that is associated with vision in D. melanogaster [90]–[92], but also has been shown to affect olfaction; mutants defective in norpA exhibited impaired olfactory capabilities [93]. Phospholipase C has been documented in the homogenate of pheromone-sensitive sensilla of the silk moth, Antheraea polyphemus [94], and has been suggested as having a role in olfactory signal transduction in another moth, Spodoptera littoralis [95].…”

Pheromone Transduction in Moths

“…The enzyme PLCβ was cloned and in situ hybridizations showed that PLCβ is transcribed in adult pheromone-sensitive ORNs of the moth Spodoptera littoralis (Chouquet et al, 2010). In addition, immunocytochemical studies located a PLCβ-subtype in homogenates of isolated pheromone-sensitives sensilla of A. polyphemus (Maida et al, 2000). This PLCβ appears to mediate insect odor transduction, because odor-dose-dependently physiological concentrations and time courses of pheromones as well as general odors caused rapid, transient rises of IP 3 in antennal homogenates within several milliseconds.…”

“…It appears to serve different functions, either as activator of PKC (Figure 3) or as activator of transient receptor potential (TRP)-type ion channels (not shown). Immunocytochemical experiments provided evidence for the colocalization of phospholipase Cβ together with a PKC in trichoid sensilla of A. polyphemus (Maida et al, 2000). The PKC appears to be employed in transduction of long, adapting pheromone stimuli because in homogenates of antennal sensilla of A. polyphemus second- to minute-long incubation with pheromone increased PKC but not PKA activity (Maida et al, 2000).…”