Differential expression of two RXR/ultraspiracle isoforms during the life cycle of the hemimetabolous insect Blattella germanica (Dictyoptera, Blattellidae)

Maestro, Óscar Caso; Cruz, Josefa; Pascual, Núria; Bellés, Xavier

doi:10.1016/j.mce.2005.04.004

Cited by 73 publications

(70 citation statements)

References 47 publications

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“…The B. germanica FTZ-F1 homologue was obtained by PCR using a cDNA template from 20E-treated UM-BGE-1 cells, as described by Maestro et al (2005). Degenerate primers were designed on the basis of conserved DBD sequences of insect homologues, as follows: forward, 5Ј-TGYGARTC-NTGYAARGG-3Ј; reverse, 5Ј-AR-RCAYTTYTGRAANCGRCA-3Ј.…”

Section: Cloning Of Bgftz-f1 Cdnamentioning

confidence: 99%

“…Given the extremely low expression levels of transcription factors, a semiquantitative RT-PCR/Southern blot approach was used (Maestro et al, 2005;Mané-Padrós et al, 2008). Total RNA was extracted from different tissues using the GenElute Mammalian Total RNA kit (Sigma).…”

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

“…Total RNA was extracted from different tissues using the GenElute Mammalian Total RNA kit (Sigma). A sample of 0.3-1 g of each RNA preparation was used for cDNA synthesis, as previously described (Maestro et al, 2005). For BgFTZ-F1 mRNA detection, cDNA samples were subjected to PCR amplification with several cycles within the linear range of amplification at 94°C for 30 sec, 64°C for 30 sec, and 72°C for 45 sec.…”

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

“…For BgFTZ-F1 mRNA detection, cDNA samples were subjected to PCR amplification with several cycles within the linear range of amplification at 94°C for 30 sec, 64°C for 30 sec, and 72°C for 45 sec. Primers used for BgFTZ-F1 detection were as follows: forward (BgFTZ-F1-F5), 5Ј-TAT-AGTCTACACAGCGAATCAGA-3Ј and reverse (FTZ-F1-R5), 5Ј-GTA-CATCGGGCCGAATTTGTTTCT-3Ј; primers used for the amplification of the different nuclear receptors were as described in Cruz et al (2006Cruz et al ( , 2007, Maestro et al (2005), and Mané-Padrós et al (2008). As a reference, the actin5C transcript of B. germanica was amplified by PCR using the same cDNA template samples (Maestro et al, 2005).…”

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

“…During the last years, and with the aim of finding regularities as well as differences in ecdysteroid action between both types of insect development, we have been characterizing the 20E-triggered genetic hierarchy of transcription factors in the German cockroach Blattella germanica, as model of hemimetabolous species. We have identified the functional 20E heterodimeric receptor, namely BgEcR-A, and two isoforms of RXR/USP, BgRXR-S, and BgRXR-L (Maestro et al, 2005;Cruz et al, 2006), as well as five isoforms of the early transcription factor E75 (Mané-Padrós et al, 2008) and three isoforms of the early-late HR3 (Cruz et al, 2007). In B. germanica, whereas BgEcR-A and BgRXR are expressed in a housekeeping-like pattern during the last two nymphal instars, BgE75 and BgHR3 isoforms are activated and repressed sequentially in response to the periodic pulse of circulating ecdysteroids (Fig.…”

Section: Introductionmentioning

confidence: 99%

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Nuclear receptor BgFTZ‐F1 regulates molting and the timing of ecdysteroid production during nymphal development in the hemimetabolous insect Blattella germanica

Cruz¹,

Nieva²,

Mané-Padrós³

et al. 2008

Developmental Dynamics

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Section: Cloning Of Bgftz-f1 Cdnamentioning

confidence: 99%

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

Section: Rt-pcr/southern Blot Analysesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Nuclear receptor BgFTZ‐F1 regulates molting and the timing of ecdysteroid production during nymphal development in the hemimetabolous insect Blattella germanica

Cruz¹,

Nieva²,

Mané-Padrós³

et al. 2008

Developmental Dynamics

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Silencing allatostatin expression using double‐stranded RNA targeted to preproallatostatin mRNA in the German cockroach

Maestro

Bellés

2006

Arch Insect Biochem Physiol

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YXFGL-NH(2) family allatostatins (ASTs) were isolated from cockroach brain extracts based on their capacity to inhibit juvenile hormone (JH) biosynthesis in corpora allata (CA) incubated in vitro. Subsequently, the inhibitory activity of synthetic ASTs was demonstrated experimentally, although these peptides were shown to be active as JH inhibitors only in cockroaches, crickets, and termites. Here, we sought to examine whether ASTs are true physiological regulators of JH synthesis. To this end, we used RNA interference methodologies and the cockroach Blattella germanica as a model. Treatments with double-stranded RNA targeting the allatostatin gene in females of B. germanica produced a rapid and long-lasting reduction in mRNA and peptide levels in both brain and midgut during the reproductive cycle. Nevertheless, while brain AST levels were reduced approximately 70-80%, JH synthesis did not increase in any of the age groups tested.

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DNA‐binding properties of Drosophila ecdysone receptor isoforms and their modification by the heterodimerization partner ultraspiracle

Braun

Azoitei

Spindler-Barth

2009

Arch Insect Biochem Physiol

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Transcriptional activity of ecdysone receptor (EcR) isoforms varies considerably and is modified further by the heterodimerization partner and hormone treatment. To investigate whether differences in DNA binding of receptor complexes are responsible for these variations in transcriptional activity, interaction of Drosophila EcR isoforms, and variants of Ultraspiracle (Usp), the orthologue of RXR, with the ecdysone response elements (EcRE) hsp 27, PAL-1, and DR-1, were determined by electrophoretic mobility shift assays. Receptor proteins were expressed in vertebrate cells (CHO-K1) in order to rule out an influence of endogenous receptor proteins. In the absence of a heterodimerization partner, weak DNA binding of EcR was detected even without hormone with EcR-A and -B1, but not EcR-B2. In the presence of hormone, all three isoforms show increased binding to the hsp 27 EcRE. The heterodimerization partner Usp increased DNA binding considerably. The hormone effect of heterodimers is more pronounced with both EcR-B isoforms compared to EcR-A. Two specific bands were obtained for EcR-A and B1 but only one band is visible with EcR-B2. Deletion of the C-domain of Usp still allows basal DNA binding of the heterodimer, but in contrast to full-length Usp, addition of hormone decreases the intensity of the retarded receptor band of all EcR isoforms and the EcREs hsp27 and DR-1 considerably, whereas interaction with the EcRE PAL-1 is only slightly affected. Synergistic effects on transcriptional activity are associated with the formation of different receptor DNA-complexes observed with 1xhsp27 and 3xhsp27. Comparison of DNA-binding properties of EcR isoforms and EcR/Usp heterodimers revealed that binding of receptor complexes to hsp 27 EcRE is dependent on the AB domain of EcR and the AB-, C-, and D-domains of the heterodimerization partner. Interaction with the hsp 27 EcRE correlates neither with ligand binding nor with transcriptional activity of the various receptor complexes. We, therefore, conclude that the different receptor functions are regulated separately, for example, by interaction with co-modulators or post-transcriptional modifications.

show abstract

Differential expression of two RXR/ultraspiracle isoforms during the life cycle of the hemimetabolous insect Blattella germanica (Dictyoptera, Blattellidae)

Cited by 73 publications

References 47 publications

Nuclear receptor BgFTZ‐F1 regulates molting and the timing of ecdysteroid production during nymphal development in the hemimetabolous insect Blattella germanica

Nuclear receptor BgFTZ‐F1 regulates molting and the timing of ecdysteroid production during nymphal development in the hemimetabolous insect Blattella germanica

Silencing allatostatin expression using double‐stranded RNA targeted to preproallatostatin mRNA in the German cockroach

DNA‐binding properties of Drosophila ecdysone receptor isoforms and their modification by the heterodimerization partner ultraspiracle

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