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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.


Developmental profiles of BmE75 in prothoracic glands.(A-A”) Developmental profiles of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) in the prothoracic glands from day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2). L, instar; D, day; EW, early wandering; LW, late wandering; PP, prepupal; P, pupa. (B-B’) Developmental profiles of protein levels of BmE75A, BmE75B and BmE75C in the prothoracic glands from L5D2 to PP2. The full-length blots are presented in Supplementary Figure 1. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). Developmental profile of ecdysteroid titer in the hemolymph measured by radioimmunoassay from L5D2 to PP2.
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f4: Developmental profiles of BmE75 in prothoracic glands.(A-A”) Developmental profiles of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) in the prothoracic glands from day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2). L, instar; D, day; EW, early wandering; LW, late wandering; PP, prepupal; P, pupa. (B-B’) Developmental profiles of protein levels of BmE75A, BmE75B and BmE75C in the prothoracic glands from L5D2 to PP2. The full-length blots are presented in Supplementary Figure 1. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). Developmental profile of ecdysteroid titer in the hemolymph measured by radioimmunoassay from L5D2 to PP2.

Mentions: The prothoracic glands produce and secrete ecdysone, the immediate precursor of 20E during insect larval development26. From day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2), the developmental expression profiles of all three BmE75 isoforms in the prothoracic glands were examined at both mRNA and protein levels. BmE75A mRNA level is under detectable during the feeding larval stages, and reaches a small peak on L5D7 and a very high peak on PP1 and PP2 (Fig. 4A). BmE75B mRNA level is under detectable during the feeding larval stages and the wandering stages, and reaches a peak on PP1 and PP2 (Fig. 4A’). Differently, BmE75C is continuously expressed from L5D2 and PP2 with relatively high expression on L5D7 and PP2 (Fig. 4A”).


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)

Developmental profiles of BmE75 in prothoracic glands.(A-A”) Developmental profiles of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) in the prothoracic glands from day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2). L, instar; D, day; EW, early wandering; LW, late wandering; PP, prepupal; P, pupa. (B-B’) Developmental profiles of protein levels of BmE75A, BmE75B and BmE75C in the prothoracic glands from L5D2 to PP2. The full-length blots are presented in Supplementary Figure 1. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). Developmental profile of ecdysteroid titer in the hemolymph measured by radioimmunoassay from L5D2 to PP2.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Developmental profiles of BmE75 in prothoracic glands.(A-A”) Developmental profiles of mRNA levels of BmE75A (A), BmE75B (A’) and BmE75C (A”) in the prothoracic glands from day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2). L, instar; D, day; EW, early wandering; LW, late wandering; PP, prepupal; P, pupa. (B-B’) Developmental profiles of protein levels of BmE75A, BmE75B and BmE75C in the prothoracic glands from L5D2 to PP2. The full-length blots are presented in Supplementary Figure 1. (B’) Quantification of BmE75A, BmE75B and BmE75C protein levels in (B). Developmental profile of ecdysteroid titer in the hemolymph measured by radioimmunoassay from L5D2 to PP2.
Mentions: The prothoracic glands produce and secrete ecdysone, the immediate precursor of 20E during insect larval development26. From day 2 of the 5th larval instar (L5D2) to day 2 of the prepupal stage (PP2), the developmental expression profiles of all three BmE75 isoforms in the prothoracic glands were examined at both mRNA and protein levels. BmE75A mRNA level is under detectable during the feeding larval stages, and reaches a small peak on L5D7 and a very high peak on PP1 and PP2 (Fig. 4A). BmE75B mRNA level is under detectable during the feeding larval stages and the wandering stages, and reaches a peak on PP1 and PP2 (Fig. 4A’). Differently, BmE75C is continuously expressed from L5D2 and PP2 with relatively high expression on L5D7 and PP2 (Fig. 4A”).

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.