|Wang, Amy - AVENTIS|
|Nabli, Henda - UNIV OF MISSOURI|
|Wittmeyer, Jennifer - UNIV OF MISSOURI|
Submitted to: Congress on In Vitro Biology
Publication Type: Abstract Only
Publication Acceptance Date: March 10, 2002
Publication Date: June 10, 2002
Citation: Goodman, C.L., Wang, A., Mcintosh, A.H., Nabli, H., Wittmeyer, J. 2002. Development and testing of insect cell lines for neuronal characteristics. Congress on In Vitro Biology. 38:16A. Technical Abstract: Few continuously replicating insect cell lines with neuronal characteristics have been identified to date. Cell lines with these characteristics can be used in physiological studies for understanding insect neuronal growth and development, as well as toxicity studies for evaluating biorational insecticides. The goal of this project was to develop and identify cell lines with neuronal characteristics for the latter use. Project objectives included: (1) the establishment of new cell cultures from insect nervous tissues; (2) the use of biochemical methods to screen newly and previously established cell lines for the presence of nervous tissue-specific proteins, including functional octopamine receptors. New cell lines were established from selected tissues of budworms, Heliothis virescens, and bollworms, Helicoverpa zea, including larval ventral nerve cords (4 lines) and midgut tissues (1 line), as well as embryonic tissues (10 lines). For characterization, immunocytochemical studies using an antibody to horseradish peroxidase (anti-HRP) were performed based on Jan and Jan's observation that anti-HRP specifically recognizes neuronal tissues (Proc. Natl. Acad. Sci. 79:2700, 1982). A total of 32 cell lines were subjected to immuostaining using an anti-HRP- FITC conjugate, with 11 cell lines being specifically recognized (including 1 neuronal line). Additionally, selected cell lines were tested for cAMP elevation in the presence of octopamine to identify those with functional octopamine receptors. Of the cell lines examined, only 2 responded with statistically higher levels of cAMP when exposed to octopamine (i.e, a new cell line from neuronal tissues of lepidopteran larvae and a previously established cell line from embryonic tissues of hymenopteran eggs).