Regulation of ovarian ecdysteroid production in the housefly, Musca domestica

Highly purified Musca ecdysteroidogenin (ESG) was obtained by extracting housefly heads with an acid-ethanol-urea procedure followed by desalting on a Sephadex G25F column and fractionation on a Sephadex G50F column. Further fractionation was done on a semi-preparative C18 reverse-phase HPLC column...

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Veröffentlicht in:Archives of insect biochemistry and physiology 1997, Vol.35 (1/2), p.135-148
Hauptverfasser: Adams, T.S, Gerst, J.W, Masler, E.P
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description Highly purified Musca ecdysteroidogenin (ESG) was obtained by extracting housefly heads with an acid-ethanol-urea procedure followed by desalting on a Sephadex G25F column and fractionation on a Sephadex G50F column. Further fractionation was done on a semi-preparative C18 reverse-phase HPLC column with an acetonitrile gradient in 0.1% trifluoroacetic acid in water. Ecdysteroidogenic activity eluted from the column with 35% acetonitrile. I-125 size exclusion HPLC revealed activity in a fraction with a molecular mass of 8.1 kDa. ESG from the C18 column at a concentration of 1.1 micrograms/fly stimulated ovarian development to late vitellogenesis when injected into flies without the corpus allatum-cardiacum complex and stimulated maximal ovarian ecdysteroid production in vitro at a concentration of 40 micrograms/microliter. Concentrations of ESG outside these ranges were less effective. Ovaries produced about 500 pg each of ecdysone, 20-hydroxyecdysone, and makisterone A when exposed to ESG in vitro. However, control ovaries produced about a tenth the amount of ecdysone and 20-hydroxyecdysone and half the amount of makisterone A. An exposure of 4 h or more to ESG was required by ovaries before they would make ecdysteroid, with the greatest rate of ecdysteroid production occurring in the 12-24 h interval of exposure. Ecdysteroid production ceased from 36-50 h after ESG was removed from the medium. A juvenile hormone analog (JHA) did not stimulate ovarian ecdysteroid production, but ovaries from flies without the corpus allatum-cardiacum complex required JHA to produce high levels of ecdysteroid during incubation with ESG. ESG activity was recovered from both male and female heads
doi_str_mv 10.1002/(SICI)1520-6327(1997)35:1/2<135::AID-ARCH12>3.0.CO;2-C
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Insect Biochem. Physiol</addtitle><description>Highly purified Musca ecdysteroidogenin (ESG) was obtained by extracting housefly heads with an acid-ethanol-urea procedure followed by desalting on a Sephadex G25F column and fractionation on a Sephadex G50F column. Further fractionation was done on a semi-preparative C18 reverse-phase HPLC column with an acetonitrile gradient in 0.1% trifluoroacetic acid in water. Ecdysteroidogenic activity eluted from the column with 35% acetonitrile. I-125 size exclusion HPLC revealed activity in a fraction with a molecular mass of 8.1 kDa. ESG from the C18 column at a concentration of 1.1 micrograms/fly stimulated ovarian development to late vitellogenesis when injected into flies without the corpus allatum-cardiacum complex and stimulated maximal ovarian ecdysteroid production in vitro at a concentration of 40 micrograms/microliter. Concentrations of ESG outside these ranges were less effective. Ovaries produced about 500 pg each of ecdysone, 20-hydroxyecdysone, and makisterone A when exposed to ESG in vitro. However, control ovaries produced about a tenth the amount of ecdysone and 20-hydroxyecdysone and half the amount of makisterone A. An exposure of 4 h or more to ESG was required by ovaries before they would make ecdysteroid, with the greatest rate of ecdysteroid production occurring in the 12-24 h interval of exposure. Ecdysteroid production ceased from 36-50 h after ESG was removed from the medium. A juvenile hormone analog (JHA) did not stimulate ovarian ecdysteroid production, but ovaries from flies without the corpus allatum-cardiacum complex required JHA to produce high levels of ecdysteroid during incubation with ESG. 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Insect Biochem. Physiol</addtitle><date>1997</date><risdate>1997</risdate><volume>35</volume><issue>1/2</issue><spage>135</spage><epage>148</epage><pages>135-148</pages><issn>0739-4462</issn><eissn>1520-6327</eissn><abstract>Highly purified Musca ecdysteroidogenin (ESG) was obtained by extracting housefly heads with an acid-ethanol-urea procedure followed by desalting on a Sephadex G25F column and fractionation on a Sephadex G50F column. Further fractionation was done on a semi-preparative C18 reverse-phase HPLC column with an acetonitrile gradient in 0.1% trifluoroacetic acid in water. Ecdysteroidogenic activity eluted from the column with 35% acetonitrile. I-125 size exclusion HPLC revealed activity in a fraction with a molecular mass of 8.1 kDa. ESG from the C18 column at a concentration of 1.1 micrograms/fly stimulated ovarian development to late vitellogenesis when injected into flies without the corpus allatum-cardiacum complex and stimulated maximal ovarian ecdysteroid production in vitro at a concentration of 40 micrograms/microliter. Concentrations of ESG outside these ranges were less effective. Ovaries produced about 500 pg each of ecdysone, 20-hydroxyecdysone, and makisterone A when exposed to ESG in vitro. However, control ovaries produced about a tenth the amount of ecdysone and 20-hydroxyecdysone and half the amount of makisterone A. An exposure of 4 h or more to ESG was required by ovaries before they would make ecdysteroid, with the greatest rate of ecdysteroid production occurring in the 12-24 h interval of exposure. Ecdysteroid production ceased from 36-50 h after ESG was removed from the medium. A juvenile hormone analog (JHA) did not stimulate ovarian ecdysteroid production, but ovaries from flies without the corpus allatum-cardiacum complex required JHA to produce high levels of ecdysteroid during incubation with ESG. ESG activity was recovered from both male and female heads</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><doi>10.1002/(SICI)1520-6327(1997)35:1/2&lt;135::AID-ARCH12&gt;3.0.CO;2-C</doi><tpages>14</tpages></addata></record>
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ispartof Archives of insect biochemistry and physiology, 1997, Vol.35 (1/2), p.135-148
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subjects 20-hydroxyecdysone
ALLATECTOMY
ALPHA-ECDYSONE
CARDIECTOMY
CONTROL HORMONAL
CONTROLE HORMONAL
CORPORA ALLATA
CORPORA CARDIACA
ecdysone
ECDYSTEROIDOGENESIS
ECDYSTEROIDOGENIN
ECDYSTERONE
EDNH
EGG DEVELOPMENT NEUROSECRETORY HORMONE
ESTEROIDES
HORMONAL CONTROL
HORMONAS
HORMONAS DE LA MUDA
HORMONAS JUVENILES
HORMONE
HORMONE DE MUE
HORMONE JUVENILE
HORMONES
juvenile hormone
JUVENILE HORMONES
MAKISTERON A
makisterone A
METABOLISME DES STEROIDES
METABOLISMO DE ESTEROIDES
METHOPRENE
METOPRENO
MOULTING HORMONES
MUSCA DOMESTICA
NERVOUS SYSTEM
NEUROHORMONES
OVAIRE
OVARIAN DEVELOPMENT
OVARIES
OVARIOS
ovary
SISTEMA NERVIOSO
STEROID METABOLISM
STEROIDE
STEROIDOGENESIS
STEROIDS
SYSTEME NERVEUX
title Regulation of ovarian ecdysteroid production in the housefly, Musca domestica
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