Patterns of MHR3 expression in the epidermis during a larval molt of the tobacco hornworm Manduca sexta

Rosalie E. Langelan; Jeffrey E. Fisher; Kiyoshi Hiruma; Subba Reddy Palli; Lynn M. Riddiford

doi:10.1006/dbio.2000.9895

Patterns of MHR3 expression in the epidermis during a larval molt of the tobacco hornworm Manduca sexta

Rosalie E. Langelan, Jeffrey E. Fisher, Kiyoshi Hiruma, Subba Reddy Palli, Lynn M. Riddiford

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34 Scopus citations

Abstract

MHR3, an ecdysone-induced transcription factor, was shown to appear in the abdominal epidermis of the tobacco hornworm Manduca sexta in a pattern-specific manner as the 20-hydroxyecdysone (20E) titer rises for the larval molt. The crochet epidermis that forms the hooked setae on the proleg is first to show MHR3 mRNA and protein followed sequentially by the spiracle, the dorsal intrasegmental annuli, the interannular regions, and finally the trichogen and tormogen cells. The protein appears in the nuclei about 8 h before the onset of cuticle formation, is present during the outgrowth of the setae, and disappears after epicuticle formation. In vitro studies showed that MHR3 mRNA induction in the crochet epidermis by 20E was more sensitive (EC₅₀ = 10⁻⁶ M; 50% induction by 2 h exposure to 4 x 10⁻⁶ M 20E) and did not require protein synthesis for maximal accumulation compared to the dorsal epidermis. The ecdysone receptor complex is present in both tissues at the outset of the molt and therefore is not a determining factor in these responses. Thus, in addition to the ecdysone receptor complex, region-specific factors govern both sensitivity and timing of responsiveness of MHR3 to 20E to ensure that this transcription factor will be present when needed for its differentiative role. (C) 2000 Academic Press.

Original language	English
Pages (from-to)	481-494
Number of pages	14
Journal	Developmental Biology
Volume	227
Issue number	2
DOIs	https://doi.org/10.1006/dbio.2000.9895
State	Published - Nov 15 2000

Bibliographical note

Funding Information:
We thank Dr. James S. Warren and Professor Lawrence Gilbert, University of North Carolina, for the HPLC-RIA analysis of the ecdysteroids present in the hemolymph; Dr. Sho Sakurai, Ka-nazawa University, for the O-6 antibody; Dr. Wendy Smith, North-eastern University, for help with the RIA protocol and for providing protein A for the assay; Dr. Takeshi Matsumoto, Daicel Chemical Co., for 20E; Drs. David Champlin and Que Lan for the affinity-purified antiMHR3 antibody; Ta Deng for assistance in some of the immunocytochemistry; and Drs. James Truman and Margrit Schu-biger for critical comments on the manuscript. The work was supported by grants from NIH (AI 12495) and NSF (IBN 95-14187 and IBN 98-17339).

Keywords

20-hydroxyecdysone
Abdominal epidermis
MHR3
Manduca sexta
Transcription factor patterning

ASJC Scopus subject areas

Molecular Biology
Developmental Biology
Cell Biology

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title = "Patterns of MHR3 expression in the epidermis during a larval molt of the tobacco hornworm Manduca sexta",

abstract = "MHR3, an ecdysone-induced transcription factor, was shown to appear in the abdominal epidermis of the tobacco hornworm Manduca sexta in a pattern-specific manner as the 20-hydroxyecdysone (20E) titer rises for the larval molt. The crochet epidermis that forms the hooked setae on the proleg is first to show MHR3 mRNA and protein followed sequentially by the spiracle, the dorsal intrasegmental annuli, the interannular regions, and finally the trichogen and tormogen cells. The protein appears in the nuclei about 8 h before the onset of cuticle formation, is present during the outgrowth of the setae, and disappears after epicuticle formation. In vitro studies showed that MHR3 mRNA induction in the crochet epidermis by 20E was more sensitive (EC50 = 10−6 M; 50% induction by 2 h exposure to 4 x 10−6 M 20E) and did not require protein synthesis for maximal accumulation compared to the dorsal epidermis. The ecdysone receptor complex is present in both tissues at the outset of the molt and therefore is not a determining factor in these responses. Thus, in addition to the ecdysone receptor complex, region-specific factors govern both sensitivity and timing of responsiveness of MHR3 to 20E to ensure that this transcription factor will be present when needed for its differentiative role. (C) 2000 Academic Press.",

keywords = "20-hydroxyecdysone, Abdominal epidermis, MHR3, Manduca sexta, Transcription factor patterning",

author = "Langelan, {Rosalie E.} and Fisher, {Jeffrey E.} and Kiyoshi Hiruma and Palli, {Subba Reddy} and Riddiford, {Lynn M.}",

note = "Funding Information: We thank Dr. James S. Warren and Professor Lawrence Gilbert, University of North Carolina, for the HPLC-RIA analysis of the ecdysteroids present in the hemolymph; Dr. Sho Sakurai, Ka-nazawa University, for the O-6 antibody; Dr. Wendy Smith, North-eastern University, for help with the RIA protocol and for providing protein A for the assay; Dr. Takeshi Matsumoto, Daicel Chemical Co., for 20E; Drs. David Champlin and Que Lan for the affinity-purified antiMHR3 antibody; Ta Deng for assistance in some of the immunocytochemistry; and Drs. James Truman and Margrit Schu-biger for critical comments on the manuscript. The work was supported by grants from NIH (AI 12495) and NSF (IBN 95-14187 and IBN 98-17339).",

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doi = "10.1006/dbio.2000.9895",

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AU - Langelan, Rosalie E.

AU - Fisher, Jeffrey E.

AU - Hiruma, Kiyoshi

AU - Palli, Subba Reddy

AU - Riddiford, Lynn M.

N1 - Funding Information: We thank Dr. James S. Warren and Professor Lawrence Gilbert, University of North Carolina, for the HPLC-RIA analysis of the ecdysteroids present in the hemolymph; Dr. Sho Sakurai, Ka-nazawa University, for the O-6 antibody; Dr. Wendy Smith, North-eastern University, for help with the RIA protocol and for providing protein A for the assay; Dr. Takeshi Matsumoto, Daicel Chemical Co., for 20E; Drs. David Champlin and Que Lan for the affinity-purified antiMHR3 antibody; Ta Deng for assistance in some of the immunocytochemistry; and Drs. James Truman and Margrit Schu-biger for critical comments on the manuscript. The work was supported by grants from NIH (AI 12495) and NSF (IBN 95-14187 and IBN 98-17339).

PY - 2000/11/15

Y1 - 2000/11/15

N2 - MHR3, an ecdysone-induced transcription factor, was shown to appear in the abdominal epidermis of the tobacco hornworm Manduca sexta in a pattern-specific manner as the 20-hydroxyecdysone (20E) titer rises for the larval molt. The crochet epidermis that forms the hooked setae on the proleg is first to show MHR3 mRNA and protein followed sequentially by the spiracle, the dorsal intrasegmental annuli, the interannular regions, and finally the trichogen and tormogen cells. The protein appears in the nuclei about 8 h before the onset of cuticle formation, is present during the outgrowth of the setae, and disappears after epicuticle formation. In vitro studies showed that MHR3 mRNA induction in the crochet epidermis by 20E was more sensitive (EC50 = 10−6 M; 50% induction by 2 h exposure to 4 x 10−6 M 20E) and did not require protein synthesis for maximal accumulation compared to the dorsal epidermis. The ecdysone receptor complex is present in both tissues at the outset of the molt and therefore is not a determining factor in these responses. Thus, in addition to the ecdysone receptor complex, region-specific factors govern both sensitivity and timing of responsiveness of MHR3 to 20E to ensure that this transcription factor will be present when needed for its differentiative role. (C) 2000 Academic Press.

AB - MHR3, an ecdysone-induced transcription factor, was shown to appear in the abdominal epidermis of the tobacco hornworm Manduca sexta in a pattern-specific manner as the 20-hydroxyecdysone (20E) titer rises for the larval molt. The crochet epidermis that forms the hooked setae on the proleg is first to show MHR3 mRNA and protein followed sequentially by the spiracle, the dorsal intrasegmental annuli, the interannular regions, and finally the trichogen and tormogen cells. The protein appears in the nuclei about 8 h before the onset of cuticle formation, is present during the outgrowth of the setae, and disappears after epicuticle formation. In vitro studies showed that MHR3 mRNA induction in the crochet epidermis by 20E was more sensitive (EC50 = 10−6 M; 50% induction by 2 h exposure to 4 x 10−6 M 20E) and did not require protein synthesis for maximal accumulation compared to the dorsal epidermis. The ecdysone receptor complex is present in both tissues at the outset of the molt and therefore is not a determining factor in these responses. Thus, in addition to the ecdysone receptor complex, region-specific factors govern both sensitivity and timing of responsiveness of MHR3 to 20E to ensure that this transcription factor will be present when needed for its differentiative role. (C) 2000 Academic Press.

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KW - MHR3

KW - Manduca sexta

KW - Transcription factor patterning

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Patterns of MHR3 expression in the epidermis during a larval molt of the tobacco hornworm Manduca sexta

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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