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Changes In Hemolymph During M. sexta Development

--Introduction--

 

Because Manduca sexta is a holometabolous insect that has four distinct life stages (egg, caterpillar, pupa, moth&emdash;see Life Cycle Stages), it is an excellent model for investigating changes in gene expression that occur during development. Numerous researchers are investigating a variety of aspects of development in M. sexta (for examples, see Studies Related to Hemolymph Changes During Development). Gross changes in the proteins present in hemolymph during larval development and among the larval, pupal, and moth stages are relatively easy to demonstrate using common protein analysis techniques such as determination of total protein concentration and SDS-PAGE analysis.

One of the proteins that is readily identifiable in M. sexta hemolymph is the arylphorin storage protein. This protein exists in the hemolymph as hexamer composed two glycosylated subunits of 77 and 72 kDa. The protein is made by the fat body and secreted into the hemolymph. It accumulates to high levels during the larval stages. During the pupal stage, this protein is thought to be degraded and used for its component amino acids.

 

Investigating developmental changes:

Overview.

Protocols:

Collection of M. sexta hemolymph.

Determination of protein concentration of M. sexta hemolymph.

SDS-polyacrylamide gel electrophoresis of M. sexta hemolymph.

 


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March 1999