The tridecapeptides Asn(13)-orcokinin and Val(13)-orcokinin, two known members of the orcokinin neuropeptide family native to crustaceans, and a novel octapeptide, orcomyotropin, FDAFTTGFamide, have been identified from extracts of hindguts of the crayfish Orconectes limosus using an isolated hindgut contractility bioassay, high-performance liquid chromatography, microsequencing and mass spectrometry. All three peptides display strong inotropic actions on crayfish hindguts. Orcomyotropin showed higher potency than the two orcokinins. Threshold concentration was approximately 5 × 10(−12)mol l(−1)versus 10(−10)mol l(−1) for the two orcokinins. An approximately fivefold increase in contraction amplitude was observed with 10(−9)mol l(−1) orcomyotropin and 10(−7)mol l(−1) of the orcokinins. Asn(13)- and Val(13)-orcokinin did not differ significantly with regard to their biological effects. Semi-isolated crayfish hearts and locust oviducts did not respond to the three peptides. Immunocytochemistry using antisera against Asn(13)-orcokinin and orcomyotropin showed that these neuropeptides are co-localized in approximately 80–90 neurones of the terminal abdominal ganglion that have been shown to innervate the entire hindgut muscularis via the intestinal nerve. The neurones form elaborate terminal branches preferentially on longitudinal hindgut muscles. Orcomyotropin is a novel crustacean member of the GF-amide family of myotropic and/or allatotropic neuropeptides from annelids, molluscs and insects.
REFERENCES
Audehm
U.
, Trube
A.
, Dircksen
H.
(1993
). Patterns and projections of crustacean-cardioactive-peptide-immunoreactive neurones of the terminal ganglion of crayfish.
Cell Tissue Res
272
, 473
–.Averof
M.
, Akam
M.
(1995
). Insect—crustacean relationship: insights from comparative developmental and molecular studies.
Phil. Trans. R. Soc. Lond. B
347
, 293
–.Biemann
K.
(1992
). Mass spectrometry of peptides and proteins.
Annu. Rev. Biochem
61
, 977
–.Bungart
D.
, Dircksen
H.
, Keller
R.
(1994
). Quantitative determination and distribution of the myotropic neuropeptide orcokinin in the nervous system of astacidean crustaceans.
Peptides
15
, 393
–.Bungart
D.
, Hilbich
C.
, Dircksen
H.
, Keller
R.
(1995
). Occurrence of analogs of the myotropic neuropeptide orcokinin in the shore crab Carcinus maenas: evidence for a novel neuropeptide family.
Peptides
16
, 67
–.Bungart
D.
, Kegel
G.
, Burdzik
S.
, Keller
R.
(1995
). H. DIRCKSENANDOTHERS2817 Orcokinin and orcomyotropin in crayfish hindgut Structure—activity relationships of the crustacean myotropic neuropeptide orcokinin.
Peptides
16
, 199
–.Christie
A. E.
, Lundquist
C. T.
, Nässel
D. R.
, Nusbaum
M. P.
(1997
). Two novel tachykinin-related peptides from the nervous system of the crab Cancer borealis.
J. Exp. Biol
200
, 2279
–.Dircksen
H.
, Keller
R.
(1988
). Immunocytochemical localization of CCAP, a novel crustacean cardioactive peptide, in the nervous system of the shore crab, Carcinus maenas L.
Cell Tissue Res
254
, 347
–.Dircksen
H.
, Muller
A.
, Keller
R.
(1991
). Crustacean cardioactive peptide in the nervous system of the locust, Locusta migratoria: an immunocytochemical study on the ventral nerve cord and peripheral innervation.
Cell Tissue Res
263
, 439
–.Dircksen
H.
, Skiebe
P.
, Abel
B.
, Agricola
H.
, Buchner
K.
, Muren
J. E.
, Nässel
D. R.
(1999
). Structure, distribution and biological activity of novel members of the allatostatin family in the crayfish Orconectes limosus.
Peptides
20
, 695
–.Duve
H.
, Johnsen
A. H.
, Maestro
J. L.
, Scott
A. G.
, Jaros
P. P.
, Thorpe
A.
(1997
). Isolation and identification of multiple neuropeptides of the allatostatin superfamily in the shore crab Carcinus maenas.
Eur. J. Biochem
250
, 727
–.Elekes
K.
, Florey
E.
, Cahill
M. A.
(1988
). Morphology and central synaptic connections of the efferent neurons innervating the crayfish hindgut.
Cell Tissue Res
254
, 369
–.Elofsson
R.
, Kauri
T.
, Nielsen
S. O.
, Stroemberg
J. O.
(1968
). Catecholamine-containing nerve fibres in the hindgut of the crayfish Astacus astacus L.
Experientia
24
, 1159
–.Friedrich
M.
, Tautz
D.
(1995
). Ribosomal DNA phylogeny of the major extant arthropod classes and the evolution of myriapods.
Nature
376
, 165
–.Gäde
G.
, Hoffmann
K. H.
, Spring
J. H.
(1997
). Hormonal regulation in insects: facts, gaps and future directions.
Physiol. Rev
77
, 963
–.Harada
A.
, Yoshida
M.
, Minakata
H.
, Nomoto
K.
, Muneoka
Y.
, Kobayashi
M.
(1993
). Structure and function of the molluscan myoactive tetradecapeptides.
Zool. Sci
10
, 257
–.Hunt
D. F.
, Yates
J. R. I.
, Shabanovitz
J.
, Bruns
M. E.
, Bruns
D. E.
(1989
). Amino acid sequence analysis of two mouse calbindin-D9K isoforms by tandem mass spectrometry.
J. Biol. Chem
264
, 6580
–.Hunt
D. F.
, Yates
J. R. I.
, Shabanovitz
J.
, Winston
S.
, Hauer
C. R.
(1986
). Protein sequencing by tandem mass spectrometry.
Proc. Natl. Acad. Sci. USA
83
, 6233
–.Kataoka
H.
, Toshi
A.
, Li
J. P.
, Carney
R. L.
, Schooley
D. A.
, Kramer
S. J.
(1989
). Identification of an allatotropin from adult Manduca sexta.
Science
243
, 1481
–.Keller
R.
(1992
). Crustacean neuropeptides: structures, functions and comparative aspects.
Experientia
48
, 439
–.Krajniak
K. G.
(1991
). The identification and structure—activity relations of a cardioactive FMRFamide-related peptide from the blue crab Callinectes sapidus.
Peptides
12
, 1295
–.Kuster
T.
, Staudenmann
W.
, Hughes
G. J.
, Heizmann
C. W.
(1991
). Parvalbumin isoforms in chicken muscle and thymus. Amino acid sequence analysis of muscle parvalbumin by tandem mass spectrometry.
Biochemistry
30
, 8812
–.Lange
A. B.
(1987
). Mode of action of proctolin on locust visceral muscle.
Arch. Insect Biochem. Physiol
24
, 79
–.Mercier
A. J.
, Lange
A. B.
, TeBrugge
V.
, Orchard
I.
(1997
). Evidence for proctolin-like and RFamide-like neuropeptides associated with the hindgut of the crayfish Procambarus clarkii.
Can. J. Zool
75
, 1208
–.Mercier
A. J.
, Orchard
I.
, TeBrugge
V.
, Skerrett
M.
(1993
). Isolation of two FMRFamide-related peptides from crayfish pericardial organs.
Peptides
14
, 137
–.Nieto
J.
, Veelaert
D.
, Derua
R.
, Waelkens
E.
, Cerstiaens
A.
, Coast
G.
, Devreese
B.
, Van Beeumen
J.
, Calderon
J.
, De Loof
A.
, Schoofs
L.
(1998
). Identification of one tachykinin-and two kinin-related peptides in the brain of the white shrimp, Penaeus vannamei.
Biochem. Biophys. Res. Commun
248
, 406
–.Orlov
J.
(1926
). Die Innervation des Darmes des Fluβkrebses.
Z. Mikr. Anat. Forsch
4
, 101
–.Paemen
L.
, Tips
A.
, Schoofs
L.
, Proost
P.
, Van Damme
J.
, De Loof
A.
(1991
). Lom-AG-myotropin: a novel myotropic peptide from the male accessory glands of Locusta migratoria.
Peptides
12
, 7
–.Price
D. A.
, Greenberg
M. J.
(1989
). The hunting of the FaRPs: the distribution of FMRFamide-related peptides.
Biol. Bull
177
, 198
–.Sithigorngul
P.
, Saraithongkum
W.
, Jaideechoey
S.
, Longyant
S.
, Sithigorngul
W.
(1998
). Novel FMRFamide-like neuropeptides from the eyestalk of the giant freshwater prawn Macrobrachium rosenbergii.
Comp. Biochem. Physiol
120
, 587
–.Skiebe
P.
, Schneider
H.
(1994
). Allatostatin peptides in the crab stomatogastric nervous system: inhibition of the pyloric motor pattern and distribution of allatostatin-like immunoreactivity.
J. Exp. Biol
194
, 195
–.Spittaels
K.
, Vankeerberghen
A.
, Schoofs
L.
, Proost
P.
, Van Damme
J.
, De Loof
A.
(1996
). Isolation and characterization of Locusta migratoria accessory gland myotropin I (Lom-Ag-MT-I) from the brain of the Colorado potato beetle, Leptinotarsa decemlineata.
Arch. Insect Biochem. Physiol
31
, 149
–.Stangier
J.
, Hilbich
C.
, Burdzik
S.
, Keller
R.
(1992
). Orcokinin: a novel myotropic peptide from the nervous system of the crayfish, Orconectes limosus.
Peptides
13
, 859
–.Stefanini
M.
, De Martino
C.
, Zamboni
L.
(1967
). Fixation of ejaculated spermatozoa for electron microscopy.
Nature
216
, 173
–.Trimmer
B. A.
, Kobierski
L. A.
, Kravitz
E. A.
(1987
). Purification and characterization of FMRFamide-like immunoreactive substances from the lobster nervous system: isolation and sequence analysis of two closely related peptides.
J. Comp. Neurol
266
, 16
–.Trube
A.
, Audehm
U.
, Dircksen
H.
(1994
). Crustacean cardioactive peptide-immunoreactive neurons in the ventral nervous system of crayfish.
J. Comp. Neurol
348
, 80
–.Ukena
K.
, Oumi
T.
, Matsushima
O.
, Ikeda
T.
, Fujita
T.
, Minakata
H.
, Nomoto
K.
(1995
). A novel gut tetradecapeptide isolated from the earthworm, Eisenia foetida.
Peptides
16
, 995
–.Van Harreveld
A.
(1936
). A physiological solution for freshwatercrustaceans.
Proc. Soc. Exp. Biol. Med
34
, 428
–.Veelaert
D.
, Schoofs
L.
, De Loof
A.
(1998
). Peptidergic control of the corpus cardiacum—corpora allata complex of locusts.
Int. Rev. Cytol
182
, 249
–.Veenstra
J. A.
, Costes
L.
(1999
). Isolation and identification of a peptide and its cDNA from the mosquito Aedes aegypti related to Manduca sexta allatotropin.
Peptides
20
, 1145
–.Veenstra
J. A.
, Lehman
H. K.
, Davis
N. T.
(1994
). Allatotropin is a cardioacceleratory peptide in Manduca sexta.
J. Exp. Biol
188
, 347
–.
This content is only available via PDF.
© 2000 by Company of Biologists
2000
