title: Cellular localization of bursicon using antisera against partial peptide sequences of this insect cuticle-sclerotizing neurohormone.
abstract: Bursicon is the final neurohormone released at the end of the molting cycle. It triggers the sclerotization (tanning) of the insect cuticle. Until now, its existence has been verified only by bioassays. In an attempt to identify this important neurohormone, bursicon was purified from homogenates of 2,850 nerve cords of the cockroach Periplaneta americana by using high performance liquid chromatography technology and two-dimensional gel electrophoresis. Bursicon bioactivity was found in four distinct protein spots at approximately 30 kDa between pH 5.3 and 5.9. The protein of one of these spots at pH 5.7 was subsequently microsequenced, and five partial amino acid sequences were retrieved. Evidence is presented that two of these sequences are derived from bursicon. Antibodies raised against the two sequences labeled bursicon-containing neurons in the central nervous systems of P. americana. One of these antisera labeled bursicon-containing neurons in the crickets Teleogryllus commodus and Gryllus bimaculatus, and the moth Manduca sexta. A cluster of four bilaterally paired neurons in the brain of Drososphila melanogaster was also labeled. In addition, this antiserum detected three spots corresponding to bursicon in Western blots of two-dimensional gels. The 12-amino acid sequence detected by this antiserum, thus, seems to be conserved even among species that are distantly related.
authors.affiliation: Department of Biological Sciences, Vanderbilt University, Box 1812, Station B, Nashville, TN 37235, USA. h.willi.honegger@vanderbilt.edu
authors.forename: Hans-Willi
authors.initials: HW
authors.lastname: Honegger
authors.forename: Daniel
authors.initials: D
authors.lastname: Market
authors.forename: Larry A
authors.initials: LA
authors.lastname: Pierce
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Dewey
authors.forename: Barbara
authors.initials: B
authors.lastname: Kostron
authors.forename: Melanie
authors.initials: M
authors.lastname: Wilson
authors.forename: Dennis
authors.initials: D
authors.lastname: Choi
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Klukas
authors.forename: Karen A
authors.initials: KA
authors.lastname: Mesce
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002490:Central Nervous System / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004330:Drosophila; D015180:Electrophoresis, Gel, Two-Dimensional; D017952:Ganglia, Invertebrate / Q000166:cytology / Q000378:metabolism; D006135:Gryllidae; D007106:Immune Sera; D007150:Immunohistochemistry; D007447:Invertebrate Hormones / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D007814:Larva; D018402:Manduca; D009479:Neuropeptides / Q000378:metabolism; D010526:Periplaneta; D013045:Species Specificity
chemical_list: D007106:Immune Sera; D007447:Invertebrate Hormones; D009479:Neuropeptides; C058620:crustacean cardioactive peptide; C002295:bursicon