Rapid quantification of juvenile hormones and their metabolites in insect haemolymph by liquid chromatography?mass spectrometry (LC-MS)

Westerlund, Stephanie A.; Hoffmann, Klaus H.

doi:10.1007/s00216-004-2598-x

Cited by 69 publications

(56 citation statements)

References 22 publications

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“…However, it is still unclear whether JH titer is different among aphids producing different morphs. The recent development of the LC-MS method to quantify JH titer in small insects (Westerlund & Hoffmann, 2004;Schwartzberg et al, 2008) enables us to now investigate the relationship between JH titer and reproductive-mode change. In parallel, the recent sequencing and assembly of the pea aphid (Acyrthosiphon pisum) genome opens new routes to identify the genes encoding key components of JH synthesis and its downstream regulation, which then allows us to follow their expression during the process of reproductive-mode switch (International Aphid Genomics Consortium, 2010).…”

Section: Jh-related Genesmentioning

confidence: 99%

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Juvenile hormone titre and related gene expression during the change of reproductive modes in the pea aphid

Ishikawa

Ogawa

Gotoh

et al. 2011

Insect Molecular Biology

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Most aphids show reproductive polyphenism, i.e. they alternate their reproductive modes from parthenogenesis to sexual reproduction in response to short photoperiod. Although juvenile hormone (JH) has been considered a likely candidate for regulating the transition from asexual to sexual reproduction after photoperiod sensing, there are few studies investigating the direct relationship between JH titers and the reproductive mode change. In addition, the sequencing of the pea aphid genome now allows identification of the genes involved in the JH pathway which then allows us to examine their expression levels in relation to the reproductive-mode switch. Using LC-MS in the pea aphid, JHIII titer was shown to be lower in aphids producing sexual morphs under short-days than in aphids producing parthenogenetic morphs in long-days. The expression levels of genes upstream and downstream of JH action were quantified by real-time qRT-PCR across the reproductive mode change. The expression level of JH esterase (JHE), which is responsible for JH degradation, was significantly higher in aphids reared under short-days. This suggests that the up-regulation of the JH degradation pathway may be responsible for the lower JHIII titer in aphids exposed to short-days, leading to the production of sexual morphs.

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Section: Jh-related Genesmentioning

confidence: 99%

“…The sample was stored at -20°C until analysis. Experimental details were modified from Chen et al, 2007. To quantify juvenile hormone titers in aphids, the experimental methods of LC-MS were slightly modified based on Westerlund et al, 2004 andCornette et al, 2008. Briefly, 5 …”

Section: Jh Quantificationmentioning

confidence: 99%

Juvenile hormone titre and related gene expression during the change of reproductive modes in the pea aphid

Ishikawa

Ogawa

Gotoh

et al. 2011

Insect Molecular Biology

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“…that have been developed (Westerlund and Hoffmann, 2004;Li et al, 2006). Although these have been validated for use in honey bees (Zhou et al, 2011) and are comparable in terms of sensitivity to radioimmunoassays and GC-MS (Chen et al, 2007) 1.…”

Section: Analysis Of Juvenile Hormone and Ecdysteroid Levels In Honeymentioning

confidence: 99%

Standard methods for physiology and biochemistry research inApis mellifera

Hartfelder

Bitondi

Brent

et al. 2013

Journal of Apicultural Research

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SummaryDespite their tremendous economic importance, and apart from certain topics in the field of neurophysiology such as vision, olfaction, learning and memory, honey bees are not a typical model system for studying general questions of insect physiology. The reason is their social lifestyle, which sets them apart from a "typical insect" and, during social evolution, has resulted in the restructuring of certain physiological pathways and biochemical characteristics in this insect. Not surprisingly, the questions that have attracted most attention by researchers working on honey bee physiology and biochemistry in general are core topics specifically related to social organization, such as caste development, reproductive division of labour and polyethism within the worker caste. With certain proteins playing key roles in these processes, such as the major royal jelly proteins (MRJPs), including royalactin and hexamerins in caste development, and vitellogenin in reproductive division of labour and age polyethism, a major section herein will present and discuss basic laboratory protocols for protein analyses established and standardized to address such questions in bees. A second major topic concerns endocrine mechanisms underlying processes of queen and worker development, as well as reproduction and polyethism, especially the roles of juvenile hormone and ecdysteroids. Sensitive techniques for the quantification of juvenile hormone levels circulating in haemolymph, as well as its synthesis by the corpora allata are described. Although these require certain instrumentation and a considerable degree of sophistication in the analysis procedures, we considered that presenting these techniques would be of interest to laboratories planning to specialize in such analyses. Since biogenic amines are both neurotransmitters and regulators of endocrine glands, we also present a standard method for the detection and analysis of certain biogenic amines of interest. Further questions that cross borders between individual and social physiology are related to energy metabolism and thermoregulation. Thus a further three sections are dedicated to protocols on carbohydrate quantification in body fluid, body temperature measurement and respirometry. Métodos estándar para la investigación de la fisiología y bioquímica de Apis mellifera ResumenA pesar de su enorme importancia económica, y aparte de ciertos temas en el campo de la neurofisiología, tales como la visión, el olfato, el aprendizaje y la memoria, las abejas no son un sistema modelo típico para el estudio de cuestiones generales sobre la fisiología de los insectos. La razón de ello es su forma de vida social, lo que las diferencia de un "insecto típico" y que durante la evolución social, se ha traducido en la reestructuración de ciertas vías fisiológicas y bioquímicas propias de este insecto. Como era de esperar, las preguntas que han atraído mayor atención por parte de los investigadores que trabajan en la fisiología y la bioquímica de la abeja melífera, son en general tema...

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“…The JH titres in the haemolymph of G. bimaculatus females were quantified by liquid chromatography-mass spectrometry (LC-MS) (Westerlund and Hoffmann, 2004). The experimental design followed the one described for experiment 2 with 21-24 females for each treatment.…”

Section: Experiments 5: Effects Of Mk-801 On In Vivo Jh Titres In G Bmentioning

confidence: 99%

“…The experimental design followed the one described for experiment 2 with 21-24 females for each treatment. Three and 24·h after the second injection on day·3, 20·l of haemolymph were collected per female and extracted (Westerlund and Hoffmann, 2004). The samples were separated on a C18 reverse-phased column (ReproSil-Pur ODS-3, 5·m; Dr Maisch GmbH, Germany), protected by a guard column (C18 cartridge; Phenomex, Aschaffenburg, Germany) with differing gradients of water-methanol.…”

Section: Experiments 5: Effects Of Mk-801 On In Vivo Jh Titres In G Bmentioning

confidence: 99%

Effects of the NMDA receptor antagonist MK-801 on female reproduction and juvenile hormone biosynthesis in the cricketGryllus bimaculatusand the butterflyBicyclus anynana

Geister

Lorenz

Hoffmann

et al. 2008

Journal of Experimental Biology

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SUMMARYApart from regulating insect development, juvenile hormones (JHs) play an important role in insect reproduction, where they initiate vitellogenin synthesis and regulate the uptake of yolk by the ovary. JH synthesis is a tightly regulated process controlled by neurons and peptidergic neurosecretory cells. One of the known stimulatory regulators of JH biosynthesis is glutamate, and its N-methyl-D-aspartate (NMDA) receptor has been recently found in the cockroach Diploptera punctata. In this study we demonstrate a strong reduction in reproductive output in the tropical butterfly Bicyclus anynana and the Mediterranean field cricket Gryllus bimaculatus caused by the NMDA receptor antagonist MK-801. Such inhibiting effects on reproduction could be overruled by the application of JH mimics. In G. bimaculatus, MK-801 inhibits in vitro JH biosynthesis in the corpora allata and reduces in vivo JH haemolymph titres in a dose-dependent manner. These results suggest that JH biosynthesis in the corpora allata is at least in part controlled by an NMDA receptor with Ca 2+ as a second level messenger. Based on our findings we consider NMDA receptor antagonists as important tools for manipulating juvenile hormone biosynthesis and therefore for gaining a better understanding of the mechanistic basis of reproduction.

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Rapid quantification of juvenile hormones and their metabolites in insect haemolymph by liquid chromatography?mass spectrometry (LC-MS)

Cited by 69 publications

References 22 publications

Juvenile hormone titre and related gene expression during the change of reproductive modes in the pea aphid

Juvenile hormone titre and related gene expression during the change of reproductive modes in the pea aphid

Standard methods for physiology and biochemistry research inApis mellifera

Effects of the NMDA receptor antagonist MK-801 on female reproduction and juvenile hormone biosynthesis in the cricketGryllus bimaculatusand the butterflyBicyclus anynana

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