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. 2016 Feb 16;10:5. doi: 10.3389/fncir.2016.00005

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Copyright © 2016 Bradley, Chapman, Lizbinski, Daly and Dacks.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of the HA GABA and Manduca sexta HA B receptor (MsHisClB) antibodies. (A) HA labeling in control animals where the antibody was not pre-adsorbed. HA labeling in the optic lobe which is the primary neurotransmitter used by arthropod receptor cells. (B) HA labeling in the optic lobe is abolished after the HA antibody was pre-adsorbed with a 10:1 HA to antibody solution. (C) GABA labeling remains in control animals where the antibody was not pre-adsorbed with GABA. (D) GABA labeling in the AL is abolished after the GABA antibody was pre-adsorbed with a 10:1 GABA to antibody solution. For each panel the same dorsal lateral axis is used. (E) Amino acid sequence alignment of the HA B receptor subunits of Manduca sexta (MsHB; Msex2.04603-RA), Drosophila melanogaster (DmHB; ACA13298.1), Apis mellifera (AmHB; ABG75740.1), and Nasonia vitripennis (NvHB; ACZ51422.1). Asterisks indicate sequence identity across all four species. Bold font indicates the immunogenic peptide sequence from Manduca that was used to generate the MsHisClB antibody. (F) Western blot using MsHisClB receptor antibody on Manduca brain tissue resulted in a single band at the predicted molecular weight (36 kDa) of the MsHisClB protein. (G) Frontal section of optic lobe depicting MsHisClB-ir in the lamina (as labeled by an asterisks). (H) Pre-adsorption with the immunogenic peptide sequence eliminates all labeling in the lamina. Scale bars = 50 μm. D, dorsal, L, lateral, A, anterior.