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Title: DEVELOPMENT AND PARTIAL CHARACTERIZATION OF HELIOTHINE CELL LINES FROM EMBRYONIC AND DIFFERENTIATED TISSUES

Author
item Goodman, Cynthia
item WANG, AMY - RHONE POULENC
item NABLI, HENDA - UNIV OF MO
item McIntosh, Arthur
item WITTMEYER, JENNIFER - UNIV OF MO
item Grasela, James

Submitted to: In Vitro Cellular and Developmental Biology - Animal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/24/2004
Publication Date: 4/1/2004
Citation: Goodman, C.L., Wang, A., Nabli, H., Mcintosh, A.H., Wittmeyer, J., Grasela, J.J. 2004. Development and partial characterization of heliothine cell lines from embryonic and differentiated tissues. In Vitro Cellular and Developmental Biology - Animals 40:89-94.

Interpretive Summary: The discovery and development of safe insect control compounds is an important part of food production because, without these compounds, pest insects can drastically decrease crop yields and food quality. In order to develop new compounds, researchers must be able to determine the effect that potential control compounds have both on pest insects and beneficial insects. Until recently, scientists have primarily used living insects for this purpose, although the interpretation of these results can be somewhat complicated. An alternative method for determining the effects that anti-insect compounds have on insects is to use cultures of insect cells that continuously replicate outside the insect body (known as insect cell lines) and have specific target sites that the compounds can attack. Two main advantages of developing screening tests based on insect cell lines instead of living insects is that these tests can be readily automated using computer-driven robotics (enabling thousands of compounds to be screened monthly) and cell lines can be grown or stored for an indefinite period of time (enabling tests to be performed and compared at any time). In this project, 17 cell lines were developed from insect nerve tissues, eggs, guts, or ovaries. These new cell lines were examined for nerve-like characteristics, with the following results being observed: 7 new cultures produced proteins similar to those produced in nerves and 3 new cultures had nerve-like appearances. Therefore, we have developed insect cell lines which can potentially be used by researchers in the agrochemical industry to determine the effects of anti-insect compounds on insect tissues.

Technical Abstract: Few continuously replicating insect cell lines with neural 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 in toxicity studies for evaluating potential insecticides. The goal of this project was to develop cell lines with neural characteristics for the latter use. In this article, we describe the establishment of new cell cultures from differentiated tissues (primarily neural) and undifferentiated tissues (primarily embryonic) and their characterization using a variety of techniques. New cell lines were established from selected tissues of the budworm, Heliothis virescens, and the bollworm, Helicoverpa zea, including larval ventral nerve cords (4 lines), midgut tissues (1 line), ovaries (1), and embryonic tissues (11 lines). Cell lines were characterized by morphological examination, DAF-PCR, ISSR-PCR, and anti-HRP immunostaining.