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In vitro bioassay for hypertrehalosemic hormone‐dependent trehalose biosynthesis by fat body from adult Blaberus discoidalis cockroaches
Author(s) -
Keeley Larry L.,
Sowa Sheila M.,
Hesson Aaron S.
Publication year - 1995
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.940280402
Subject(s) - trehalose , cockroach , hemolymph , biology , incubation , bioassay , in vitro , adipokinetic hormone , metabolism , biochemistry , in vivo , hormone , medicine , endocrinology , fat body , microbiology and biotechnology , gene , genetics , ecology
An in vitro bioassay suitable for routine use to investigate hypertrehalosemic hormone (HTH)‐dependent trehalose biosynthesis was developed for the cockroach fat body. Blaberus discoidalis fat bodies were isolated and divided so that eight matched pieces from a single tissue could be compared for multiple control and experimental treatments. Optimum incubation conditions and the properties of HTH‐dependent trehalose synthesis were determined. Dose‐response studies determined an EC 50 of 0.044 nM HTH for male fat body and 0.16 nM HTH for female tissue. HTH increased trehalose production by male fat body 3‐fold compared to only a 67% maximum increase by the female tissue, and only the male tissue was used in subsequent studies. Fat body required only 5‐min exposure to HTH for maximum trehalose production for 1 h. Trehalose synthesis was inhibited by ≥ 15 mM trehalose in the incubation medium. The fat body showed a developmental increase in trehalose synthesis in vitro that was reflected by hemolymph trehalose in vivo. Basal and HTH‐related trehalose synthesis were low between days 0 and 10, increased 3‐fold by day 20, and were high thereafter. These studies have established baseline data for future investigations to identify the signal transduction mechanisms involved in HTH regulation of fat body metabolism. © 1995 Wiley‐Liss, Inc.