Midgut and fatbody mitochondrial transhydrogenase activities during larval-pupal development of the tobacco hornworm, Manduca sexta

Vandock, Kurt P.; Drummond, Christopher A.; Smith, Stan L.; Fioravanti, Carmen F.

doi:10.1016/j.jinsphys.2010.01.008

Cited by 6 publications

(22 citation statements)

References 19 publications

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“…This stage was chosen because previous research has shown the biochemical activity and de novo synthesis of transhydrogenase to be highest at this particular stage of fifth larval instar development in midgut. 8 After cDNA synthesis using random or oligo dT primers, PCR was carried out, electrophoresis of the resulting fragments showed a product between 200 and 300 bp (Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…[8][9][10] Indeed, careful experimentation has shown that fatbody and midgut tissues display increased levels Agarose gel of pCr products of M. sexta transhydrogenase fragment using degenerate primers. notes: Degenereate primers (5′ TAYCAYCANGTNTGGGGN3′ and 5′ rAACATrTCNArCATNCK3) designed against the conserved polypeptide sequences YhTVWG and rMLDMFK (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…9,10 The developmental significance of these transhydrogenations and the role(s) they may play in postembryonic development was evaluated. 8 This transhydrogenase appears to be physiologically linked to the developmentally necessary NADPH-requiring mitochondrial E20-M.…”

Section: Introductionmentioning

confidence: 99%

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Sequence and Structure of the Tobacco Hornworm, Manduca sexta, Transhydrogenase Gene

et al. 2012

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Abstract:The reversible mitochondrial transhydrogenase in Manduca sexta has been characterized and is involved with endocrinemediated post-embryonic larval development in this model insect. While biochemical in vivo and in vitro studies have been accomplished, robust molecular studies of the transhydrogenase have not been possible due to deficient genomic data. In the present study, using a combination of degenerate oligonucleotide primers and raw genomic data, we have determined the structure and sequence of the transhydrogenase gene from the model insect Manduca sexta. The encoded protein is highly similar to other transhydrogenase proteins and this sequence is the first lepidopteran sequence reported to date.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Sequence and Structure of the Tobacco Hornworm, Manduca sexta, Transhydrogenase Gene

et al. 2012

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“…Mitochondria from 5 th instar M sexta (Day 4) were extracted by a procedure similar to that described by Vandock et al (2010). Insects were immobilized on ice prior to isolation of the midguts.…”

Section: Methodsmentioning

confidence: 99%

“…Preliminary developmental studies suggest that the role of M. sexta transhydrogenase in the energy-linked formation of NADPH may serve as a source of this reducing equivalent which in turn is required for the completion of post-embryonic development (Vandock et al 2010):…”

Section: Introductionmentioning

confidence: 99%

Effects of 20-Hydroxyecdysone on the Reversible Mitochondrial Transhydrogenase in the Tobacco Hornworm,Manduca sexta

Vandock

Perregaux

Consiglio

2013

Journal of Insect Science

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The reversible, mitochondrial membrane-associated transhydrogenase from the midgut of Manduca sexta (L.) (Lepidoptera: Sphingidae) catalyzes hydride-ion transfer between NADP(H) and NAD(H). The effects of ecdysone and 20-hydroxyecdysone were evaluated and compared to both the NADH-NADP+ and NADPH-NAD+ transhydrogenations. In the direction of NADPH-formation, the developmentally significant transhydrogenations occur as non-energy- or energy-linked reactions. The energy-linked activity couples with either electron transport-dependent NADH or succinate utilization, or ATP hydrolysis by Mg++ -dependent ATPase. Upon the addition of ecdysone alone, all energy-linked reactions in the direction of NADPH formation exhibited a notable increase in activity level over the control reaction. The addition of 20-hydroxyecdysone yielded no significant increase in the activity of any of the transhydrogenations. Synergistic addition of both ecdysone and 20-hydroxyecdysone resulted in no significant effect on transhydrogenase activity. The results of this study make evident a relationship between the presence of ecdysone and 20-hydroxyecdysone on the overall activity of M. sexta midgut mitochondrial transhydrogenations. The potential mediation of the energy-linked mitochondrial transhydrogenations involved with NADPH synthesis through the developmental relationship of ecdysone and 20-hydroxyecdysone is considered.

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EFFECTS OF PLANT FLAVONOIDS ON Manduca sexta (TOBACCO HORNWORM) FIFTH LARVAL INSTAR MIDGUT AND FAT BODY MITOCHONDRIAL TRANSHYDROGENASE

Vandock

Mitchell

Fioravanti

2012

Arch Insect Biochem Physiol

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The reversible, membrane-associated transhydrogenase that catalyzes hydride-ion transfer between NADP(H) and NAD(H) was evaluated and compared to the corresponding NADH oxidase and succinate dehydrogenase activities in midgut and fat body mitochondria from fifth larval instar Manduca sexta. The developmentally significant NADPH-forming transhydrogenation occurs as a nonenergy- or energy-linked activity with energy for the latter derived from either electron transport-dependent NADH or succinate utilization, or ATP hydrolysis by Mg++-dependent ATPase. In general, the plant flavonoids examined (chyrsin, juglone, morine, quercetin, and myricetin) affected all reactions in a dose-dependent fashion. Differences in the responses to the flavonoids were apparent, with the most notable being inhibition of midgut, but stimulation of fat body transhydrogenase by morin, and myricetin as also noted for NADH oxidase and succinate dehydrogenase. Although quercetin inhibited or stimulated transhydrogenase activity depending on the origin of mitochondria, it was without effect on either midgut or fat body NADH oxidase or succinate dehydrogenase. Observed sonication-dependent increases in flavonoid inhibition may well reflect an alteration in membrane configuration, resulting in increased exposure of the enzyme systems to the flavonoids. The effects of flavonoids on the transhydrogenation, NADH oxidase, and succinate dehydrogenase reactions suggest that compounds of this nature may prove valuable in the control of insect populations by affecting these mitochondrial enzyme components.

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Midgut and fatbody mitochondrial transhydrogenase activities during larval-pupal development of the tobacco hornworm, Manduca sexta

Cited by 6 publications

References 19 publications

Sequence and Structure of the Tobacco Hornworm, Manduca sexta, Transhydrogenase Gene

Sequence and Structure of the Tobacco Hornworm, Manduca sexta, Transhydrogenase Gene

Effects of 20-Hydroxyecdysone on the Reversible Mitochondrial Transhydrogenase in the Tobacco Hornworm,Manduca sexta

EFFECTS OF PLANT FLAVONOIDS ON Manduca sexta (TOBACCO HORNWORM) FIFTH LARVAL INSTAR MIDGUT AND FAT BODY MITOCHONDRIAL TRANSHYDROGENASE

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