Rodney Kwok

University of Toronto, Toronto, Ontario, Canada

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Publications (11)19.44 Total impact

  • Rodney Kwok, Stephen S Tobe
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    ABSTRACT: The hemolymph of invertebrates contains factors that facilitate clotting as a defense mechanism for injury. However, the clotting process may impair the measurement of hormone titers by interfering with the extraction of peptides. Using hemolymph from freshwater crayfish, our results demonstrate that hemolymph clotting appears to reduce both the amount of an endogenous peptide(s) (Dippu-AST 11-like) extracted from hemolymph, as well as the amount of spiked peptide tracer ([125I]-Dippu-AST 11) recovered from hemolymph. The efficacy of peptide extraction from hemolymph was improved by collecting the hemolymph into a variety of different media prior to hemolymph coagulation. Hemolymph samples collected into media containing the anticoagulant, citrate, had the highest amount of endogenous Dippu-AST 11-like peptide extracted as well as the highest percent recovery of spiked tracer.
    Peptides 04/2006; 27(3):590-6. · 2.52 Impact Factor
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    ABSTRACT: Manduca sexta allatotropin (Manse-AT) was first isolated on the basis of its ability to stimulate production of juvenile hormone in that insect. We examined whether this neuropeptide affects corpus allatum activity and visceral muscle contraction in adult females of the earwig, Euborellia annulipes. We also assessed the presence of allatotropin-like material in tissues using immunocytochemistry. Manse-AT at 1 nM to 10 muM stimulated juvenile hormone production in vitro by glands of low activity from 2-day virgin females. In glands of high activity from 12-day mated females, 1 and 100 nM allatotropin were effective, but 10 muM was not. Similarly, hindguts of 2-day and 12-day females significantly increased in motility in vitro in response to Manse-AT. A monoclonal antibody to Manse-AT was used to demonstrate allatotropin-like material throughout the nervous system of 2-day, virgin females. Immunoreactivity was most pronounced within varicosities of the corpora cardiaca and perisympathetic organs. No immunofluorescence was observed in gut tissue. Lastly, we showed that extract of retrocerebral complexes also enhanced in vitro hindgut motility from 2-day virgin females, in a dose-dependent manner. These results indicate material similar to M. sexta allatotropin in female earwigs and that such peptides may modulate juvenile hormone biosynthesis and visceral muscle contractions. Sensitivity to the peptides may change with physiological stage.
    Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology 10/2005; 142(1):113-22. · 2.07 Impact Factor
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    ABSTRACT: Decapod crustaceans do not appear to produce juvenile hormone, but rather its immediate precursor, methyl farnesoate (MF). Both MF and its immediate precursor, farnesoic acid (FA) are produced by the mandibular organs (MO) in crustaceans. The MO are homologous to the insect corpora allata (CA), the site of juvenile hormone biosynthesis. However, the FGLamide allatostatin (ASTs) peptides, of which there are about 60 distinct forms reported from crustaceans, have previously been found to have no effect on MO activity in crustaceans. We have identified by immunocytochemistry the presence of FGLamide-like AST immunoreactivity in neurosecretory cells throughout the CNS as well as in neurohaemal structures such as the sinus gland and pericardial organs. The ASTs are likely delivered to the MO hormonally and/or by local neurohaemal release. Using MO from adult males, we have found wide variability between animals in the in vitro rates of MF and FA biosynthesis. Treatment with Dippu-ASTs has a statistically significant stimulatory effect on MF synthesis, but only in MO that are initially producing MF at lower rates. No effect on FA production was observed, suggesting that the FGLamide ASTs exert their effect on the o-methyl transferase, the enzyme responsible for the conversion of FA to MF.
    Journal of Insect Physiology 05/2005; 51(4):367-78. · 2.38 Impact Factor
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    ABSTRACT: In the subterranean termite Reticulitermes flavipes, allatostatins (ASTs) with the C-terminus Phe-Gly Leu-amide were localized by immunocytochemistry with antibody against a cockroach AST, Dippu AST-7. AST-immunoreactivity occurred in the corpus cardiacum and corpus allatum and in the lateral and medial neurosecretory cells of the brain that innervate these organs as well as in many other nerve cells of the brain. This was observed in workers, nymphs, soldiers and secondary reproductives. A radioimmunoassay, using anti-Dippu AST-11, demonstrated about 40 fmole equivalents of AST in brains of soldiers and secondary reproductives. The product of the corpora allata in this species was determined to be juvenile hormone III. Its synthesis by corpora allata of secondary reproductives, determined by in vitro radiochemical assay, was inhibited in a dose-dependent fashion by two cockroach allatostatins, Dippu AST-7 and Dippu AST-11. Thus, as in cockroaches and crickets, allatostatin-containing nerves innervate the corpora allata of this termite species and their production of juvenile hormone is inhibited by these neuropeptides.
    Journal of Insect Physiology 05/2005; 51(4):357-65. · 2.38 Impact Factor
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    ABSTRACT: The invertebrate tachykinin-related peptides (TRPs) with the conserved C-terminal sequence FX1GX2Ramide shows sequence similarity to the vertebrate tachykinins after which they are named, and are hypothesized to be ancestrally related. In this study a polyclonal antiserum generated against a locust tachykinin (LomTK I), was used to demonstrate the presence and describe the distribution of LomTK-like immnoreactivity in the CNS and gut of Rhodnius prolixus. Reverse phase high performance liquid chromatography (RP-HPLC) was used in combination with a sensitive radioimmunoassay (RIA) to demonstrate picomolar amounts of immunoreactive material in the CNS, and femptomolar amounts associated with the hindgut. Furthermore, the results from CNS extracts separated by RP-HPLC, suggest that at least two tachykinin isoforms exist in R. prolixus. A hindgut contraction assay was developed to quantify the myotropic effects of selected LomTKs on R. prolixus hindgut contraction. Both LomTK I and II caused an increase in the frequency of hindgut contractions with EC50 values of 3.6x10(-8)M and 3.8x10(-8)M for LomTK I and II, respectively.
    Peptides 02/2005; 26(1):43-51. · 2.52 Impact Factor
  • R KWOK, J RUIZHANG, S TOBE
    Journal of Insect Physiology - J INSECT PHYSIOL. 01/2005; 51(4):367-378.
  • Journal of Insect Physiology - J INSECT PHYSIOL. 01/2005; 51(4):357-365.
  • Rodney Kwok, Ian Orchard
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    ABSTRACT: We have developed a semi-intact preparation-consisting of an isolated oviduct with abdominal ganglia VII and VIII intact and attached-with which to characterize the effects on oviduct contraction, of peptides that are bath applied to CNS tissues. The work presented here offers a qualitative analysis of the central effects of SchistoFLRFamide and proctolin upon action potentials recorded from the oviducal nerves and upon oviduct contraction. In the process of this, we hope to demonstrate that a previously characterized putative CNS SchistoFLRFamide receptor [Peptides 23 (2002) 765] is a functional receptor.SchistoFLRFamide (10(-6)M), bath applied to abdominal ganglion VII, caused an increase in action potential frequencies recorded from the oviducal nerves, as well as an increase in the frequency of phasic contractions of the oviduct. Although the function of this response is not known, these results further support the possibility that the putative CNS SchistoFLRFamide receptors are functional receptors. Proctolin (10(-6)M), bath applied to abdominal ganglion VIII, altered the rhythmic bursting of action potentials recorded from the oviducal nerve and changed the appearance and cycle duration of neurogenic oviduct contractions.
    Peptides 12/2002; 23(11):1925-32. · 2.52 Impact Factor
  • Rodney Kwok, Ian Orchard
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    ABSTRACT: A putative SchistoFLRFamide receptor in CNS membrane preparations of Locusta migratoria was characterized by cold competition binding and kinetic binding assays using [125I][Y(1)]SchistoFLRFamide ([125I]YDVDHVFLRFamide) as a radioligand. Binding to this site was saturable, specific, reversible, and of high-affinity. Data fit to a single-site binding model by non-linear regression (r(2) = 0.99) estimated K(d) = 1.73 +/- 0.45 x 10(-9) M and B(max) = 49.0 +/- 12.2 fmol.mg(-1) tissue. Total binding of [125I][Y(1)]SchistoFLRFamide to membrane preparations was reduced in the presence of GTPgammaS, an indication that the putative receptor is G protein-coupled. Structure-activity studies determined that the minimum sequence required for binding was HVFLRFamide. Other aspects of the ligand receptor interaction were also examined.
    Peptides 05/2002; 23(4):765-72. · 2.52 Impact Factor
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    ABSTRACT: The presence of locustatachykinin (LomTK)-like immunoreactivity is demonstrated in the central nervous system (CNS) of Locusta migratoria with the use of a polyclonal antiserum raised against LomTK1. By developing a radioimmunoassay with the same antiserum, we have demonstrated picomolar amounts of LomTK-like material in the tissues of the central nervous system. In contrast, only femptomolar amounts of LomTK-like material are associated with the oviduct tissue. The relative amounts of the different LomTK isoforms in the brain and the abdominal ganglionic chain were examined by separating the native peptides on high-performance liquid chromatography and comparing their retention times to synthetic LomTK standards. The amounts of the different isoforms of LomTK differed between and within the two regions of the central nervous system. However, the ratios of the different isoform amounts were similar between the two regions. The myostimulatory activities of LomTKs 1 to 4 were characterized by using the locust oviduct bioassay. LomTKs 1, 2, and 3 appeared to be more efficacious than LomTK4.
    Peptides 02/1999; 20(6):687-94. · 2.52 Impact Factor
  • Comparative Biochemistry and Physiology A-molecular & Integrative Physiology - COMP BIOCHEM PHYSIOL PT A. 01/1999; 124.