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Bombyx E75 isoforms display stage- and tissue-specific responses to 20-hydroxyecdysone.

Li K, Guo E, Hossain MS, Li Q, Cao Y, Tian L, Deng X, Li S - Sci Rep (2015)

Bottom Line: At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E.At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues.In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

View Article: PubMed Central - PubMed

Affiliation: 1] Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/Guangdong Provincial Sericulture and Mulberry Engineering Research Center, College of Animal Sciences, South China Agricultural University, Guangzhou, 510642, China [2] Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

ABSTRACT
Resulted from alternative splicing of the 5' exons, the nuclear receptor gene E75 in the silkworm, Bombyx mori, processes three mRNA isoforms, BmE75A, BmE75B and BmE75C. From the early 5(th) larval instar to the prepupal stages, BmE75A mRNA and protein levels in the prothoracic glands display developmental profiles similar to ecdysteroid titer. In the fat body, mRNA levels but not protein levels of all three BmE75 isoforms correlate with ecdysteroid titer; moreover, proteins of all three BmE75 isoforms disappear at the prepupal stages, and a modified BmE75 protein with smaller molecular weight and cytoplasm localization occurs. At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E. At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues. In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

No MeSH data available.


Related in: MedlinePlus

Tissue-specific responses of BmE75 isoforms to 20E on L5D2.(A-A”) Responses of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) to 2 μM 20E or/and 10 μg/ml cycloheximide (CHX) in the prothoracic glands on day 2 of the fifth instar (L5D2). The mRNA levels were measured 2 hours after treatment in vitro. (B-B’) Response of protein levels of BmE75A, BmE75B and BmE75C to 20E or/and CHX in the L5D2 prothoracic glands 2 hours after treatment. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). (C-D’) The same as (A-B’) except the fat body was used.
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f6: Tissue-specific responses of BmE75 isoforms to 20E on L5D2.(A-A”) Responses of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) to 2 μM 20E or/and 10 μg/ml cycloheximide (CHX) in the prothoracic glands on day 2 of the fifth instar (L5D2). The mRNA levels were measured 2 hours after treatment in vitro. (B-B’) Response of protein levels of BmE75A, BmE75B and BmE75C to 20E or/and CHX in the L5D2 prothoracic glands 2 hours after treatment. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). (C-D’) The same as (A-B’) except the fat body was used.

Mentions: In the prothoracic glands, BmE75A mRNA levels increased approximately 40-fold by 20E, CHX did not affect BmE75A expression, and the 20E induction was not blocked by CHX, indicating that BmE75A is 20E primary-responsive in this tissue (Fig. 6A). 20E or CHX decreased half of BmE75B mRNA levels, while 20E and CHX together showed no effects on BmE75B expression (Fig. 6A’). Despite that BmE75C mRNA levels increased approximately 30-fold by 20E, the 20E induction was significantly blocked by CHX, indicating that BmE75C is 20E secondary-responsive in this tissue (Fig. 6A”). Meanwhile, protein levels of BmE75A, BmE75B and BmE75C increased approximately 2.2-, 1.4- and 1.7-fold by 20E, and the 20E induction was not blocked by CHX (Fig. 6B, B”).


Bombyx E75 isoforms display stage- and tissue-specific responses to 20-hydroxyecdysone.

Li K, Guo E, Hossain MS, Li Q, Cao Y, Tian L, Deng X, Li S - Sci Rep (2015)

Tissue-specific responses of BmE75 isoforms to 20E on L5D2.(A-A”) Responses of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) to 2 μM 20E or/and 10 μg/ml cycloheximide (CHX) in the prothoracic glands on day 2 of the fifth instar (L5D2). The mRNA levels were measured 2 hours after treatment in vitro. (B-B’) Response of protein levels of BmE75A, BmE75B and BmE75C to 20E or/and CHX in the L5D2 prothoracic glands 2 hours after treatment. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). (C-D’) The same as (A-B’) except the fat body was used.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4499807&req=5

f6: Tissue-specific responses of BmE75 isoforms to 20E on L5D2.(A-A”) Responses of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) to 2 μM 20E or/and 10 μg/ml cycloheximide (CHX) in the prothoracic glands on day 2 of the fifth instar (L5D2). The mRNA levels were measured 2 hours after treatment in vitro. (B-B’) Response of protein levels of BmE75A, BmE75B and BmE75C to 20E or/and CHX in the L5D2 prothoracic glands 2 hours after treatment. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). (C-D’) The same as (A-B’) except the fat body was used.
Mentions: In the prothoracic glands, BmE75A mRNA levels increased approximately 40-fold by 20E, CHX did not affect BmE75A expression, and the 20E induction was not blocked by CHX, indicating that BmE75A is 20E primary-responsive in this tissue (Fig. 6A). 20E or CHX decreased half of BmE75B mRNA levels, while 20E and CHX together showed no effects on BmE75B expression (Fig. 6A’). Despite that BmE75C mRNA levels increased approximately 30-fold by 20E, the 20E induction was significantly blocked by CHX, indicating that BmE75C is 20E secondary-responsive in this tissue (Fig. 6A”). Meanwhile, protein levels of BmE75A, BmE75B and BmE75C increased approximately 2.2-, 1.4- and 1.7-fold by 20E, and the 20E induction was not blocked by CHX (Fig. 6B, B”).

Bottom Line: At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E.At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues.In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

View Article: PubMed Central - PubMed

Affiliation: 1] Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding/Guangdong Provincial Sericulture and Mulberry Engineering Research Center, College of Animal Sciences, South China Agricultural University, Guangzhou, 510642, China [2] Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

ABSTRACT
Resulted from alternative splicing of the 5' exons, the nuclear receptor gene E75 in the silkworm, Bombyx mori, processes three mRNA isoforms, BmE75A, BmE75B and BmE75C. From the early 5(th) larval instar to the prepupal stages, BmE75A mRNA and protein levels in the prothoracic glands display developmental profiles similar to ecdysteroid titer. In the fat body, mRNA levels but not protein levels of all three BmE75 isoforms correlate with ecdysteroid titer; moreover, proteins of all three BmE75 isoforms disappear at the prepupal stages, and a modified BmE75 protein with smaller molecular weight and cytoplasm localization occurs. At the early 5(th) larval instar stage, treatment of the prothoracic glands and fat body with 20-hydroxyecdysone (20E) and/or cycloheximide (CHX) revealed that BmE75A is 20E primary-responsive at both mRNA and protein levels, while BmE75B and BmE75C exhibit various responses to 20E. At the early wandering stage, RNAi-mediated reduction of gene expression of the 20E nuclear receptor complex, EcR-USP, significantly decreased mRNA and protein levels of all three BmE75 isoforms in both tissues. In conclusion, BmE75 isoforms display stage- and tissue-specific responses to 20E at both mRNA and protein levels; moreover, they are regulated by other unknown factors at the protein level.

No MeSH data available.


Related in: MedlinePlus