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First published online August 31, 2007
Journal of Experimental Biology 210, 3245-3254 (2007)
Published by The Company of Biologists 2007
doi: 10.1242/jeb.007740
Effects of elevated ecdysteroid on tissue expression of three guanylyl cyclases in the tropical land crab Gecarcinus lateralis: possible roles of neuropeptide signaling in the molting gland
1 Department of Biology, Colorado State University, Fort Collins, CO 80523,
USA
2 Bodega Marine Laboratory, University of California-Davis, Bodega Bay, CA
94923, USA
* Author for correspondence (e-mail: don{at}lamar.colostate.edu)
Accepted 5 July 2007
Two eyestalk (ES) neuropeptides, molt-inhibiting hormone (MIH) and
crustacean hyperglycemic hormone (CHH), increase intracellular cGMP levels in
target tissues. Both MIH and CHH inhibit ecdysteroid secretion by the molting
gland or Y-organ (YO), but apparently through different guanylyl cyclase
(GC)-dependent pathways. MIH signaling may be mediated by nitric oxide
synthase (NOS) and NO-sensitive GC. CHH binds to a membrane receptor GC. As
molting affects neuropeptide signaling, the effects of ecdysteroid on the
expression of the land crab Gecarcinus lateralis ß subunit of a
NO-sensitive GC (Gl-GC-Iß), a membrane receptor GC (Gl-GC-II) and a
NO-insensitive soluble GC (Gl-GC-III) were determined. Gl-GC-Iß isoforms
differing in the absence or presence of an N-terminal 32-amino acid sequence
and Gl-GC-III were expressed at higher mRNA levels in ES ganglia, gill,
hepatopancreas, ovary and testis, and at lower levels in YO, heart and
skeletal muscle. Three Gl-GC-II isoforms, which vary in the length of
insertions (+18, +9 and +0 amino acids) within the N-terminal ligand-binding
domain, differed in tissue distribution. Gl-GC-II(+18) was expressed highly in
striated muscle (skeletal and cardiac muscles); Gl-GC-II(+9) was expressed in
all tissues examined (ES ganglia, YO, gill, hepatopancreas, striated muscles
and gonads); and Gl-GC-II(+0) was expressed in most tissues and was the
dominant isoform in ES and thoracic ganglia. ES ablation, which increased
hemolymph ecdysteroid, increased Gl-GC-II(+18) mRNA level in claw muscle.
Using real-time RT-PCR, ES ablation increased Gl-GC-Iß, Gl-GC-III and
ecdysone receptor mRNA levels in the YOs
ten-,
four- and
twofold, respectively, whereas Gl-GC-II mRNA level was unchanged. A
single injection of 20-hydroxyecdysone into intact animals transiently lowered
Gl-GC-Iß in hepatopancreas, testis and skeletal muscle, and certain
Gl-GC-II isoforms in some of the tissues. These data suggest that YO and other
tissues can modulate responses to neuropeptides by altering GC expression.
Key words: guanylyl cyclase, molting, ecdysteroid, Crustacea, Arthropoda, Y-organ, gene expression, skeletal muscle, nervous system, digestive gland, molt-inhibiting hormone, crustacean hyperglycemic hormone, ecdysone receptor, mRNA
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