Expression of muscarinic acetylcholine receptors and choline acetyltransferase enzyme in cultured antennal sensory neurons and non-neural cells of the developing mothManduca sexta

Abstract

Antennal sensory neurons of Manduca sexta emerge from epidermal cells that also give rise to sheath cells surrounding the peripheral parts of the neurons and to glial cells that enwrap the sensory axons in the antennal nerve. Reciprocal interactions between sensory neurons and glial cells are believed to aid in axon growth and guidance, but the exact nature of these interactions is not known. We investigated the possibility of cholinergic interactions in this process by locating muscarinic acetylcholine receptors (mAChRs) and choline acetyltransferase (ChAT) enzyme in cultured antennal sensory neurons and non-neural cells. ChAT and mAChRs were present in the sensory neurons from the first day in culture. Therefore, the sensory neurons are probably cholinergic, as previously suggested, but they may also be controlled by ACh. In 7-day-old cultures a subgroup of small non-neural cells with processes expressed ChAT activity, and in 14-day-old cultures non-neural cells that formed lamellipodia and scaffoldlike structures on the culture substrate were labeled with ChAT antibody. mAChR activity was detected in similar non-neural cells but only in areas surrounding the nuclei. In addition, mAChRs were found in flat lamellipodia and filopodia forming cells that were present in 1-day-old cultures and grew in size during the 2 week investigation period. These findings suggest muscarinic cholinergic interactions between the neural and non-neural cells during the development of Manduca antenna.

Cloning and expression analysis of a muscarinic cholinergic receptor from the brain of ant, Polyrhachis vicina

“…Real-time quantitative RT-PCR analysis on the brains of three castes showed that PmAchR was expressed in all tested samples, which accords with previous studies revealing that mAchRs was expressed in various insect nervous systems (Blake et al, 1993;Torkkeli et al, 2005). The lowest expression level in the brains of males may relate to male biology: males have the simplest behavior in its short life: they leave the nest, fly, and mate with a female ant before they die.…”

Pilocarpine improves recognition of nestmates in young honey bees

“…This process necessarily involves detection of odor cues through receptors on the antennae and mouthparts. In other insects, and presumably in the honey bee, cholinergic projections carry these signals to the antennal lobes and the gustatory regions of the subesophageal ganglion [14,18,19,34,37]. Cholinergic projection neurons in turn convey information about chemical signals to the lateral horn and mushroom bodies of the protocerebrum [11,41].…”

First visualization of cholinergic cells and fibers by immunohistochemistry for choline acetyltransferase of the common type in the optic lobe and peduncle complex of Octopus vulgaris

“…A number of specific ChAT antibodies have made it possible to map immunohistochemically detailed cholinergic systems mainly in mammals (Kimura et al, 1980(Kimura et al, , 1981Eckenstein and Thoenen, 1982) and partly in some nonmammalian vertebrates, including birds (Johnson and Epstein, 1986;Medina and Reiner, 1994), amphibians (Marin et al, 1997), fishes (Tavares and Hasson-Voloch, 1998;Adrio et al, 2000;Anadón et al, 2000;Pérez et al, 2000;Pombal et al, 2001;Mueller et al, 2004), and lizards (Medina et al, 1993). In invertebrates a similar approach has been used to produce antibodies against purified ChAT from invertebrate animals, though limited to animals such as flies (Takagawa and Salvaterra, 1996;Yasuyama et al, 2003), moths (Clark et al, 2005;Torkkeli et al, 2005), and worms (Duerr et al, 2008). In addition, antibodies against human placenta ChAT have been applied to study crossreactive proteins in arthropods (Rind and Leitinger, 2000;Lipovsek et al, 2003) and crustacea (Faimali et al, 2003;Gallus et al, 2006).…”