BackgroundNR2E1 (Tlx) is an orphan nuclear receptor that regulates the maintenance and self-renewal of neural stem cells, and promotes tumourigenesis. Nr2e1-null mice exhibit reduced cortical and limbic structures and pronounced retinal dystrophy. NR2E1 functions mainly as a repressor of gene transcription in association with the co-repressors atrophin-1, LSD1, HDAC and BCL11A. Recent evidence suggests that NR2E1 also acts as an activator of gene transcription. However, co-activator complexes that interact with NR2E1 have not yet been identified. In order to find potential novel co-regulators for NR2E1, we used a microarray assay for real-time analysis of co-regulator–nuclear receptor interaction (MARCoNI) that contains peptides representing interaction motifs from potential co-regulatory proteins, including known co-activator nuclear receptor box sequences (LxxLL motif).ResultsWe found that NR2E1 binds strongly to an atrophin-1 peptide (Atro box) used as positive control and to 19 other peptides that constitute candidate NR2E1 partners. Two of these proteins, p300 and androgen receptor (AR), were further validated by reciprocal pull-down assays. The specificity of NR2E1 binding to peptides in the array was evaluated using two single amino acid variants, R274G and R276Q, which disrupted the majority of the binding interactions observed with wild-type NR2E1. The decreased binding affinity of these variants to co-regulators was further validated by pull-down assays using atrophin1 as bait. Despite the high conservation of arginine 274 in vertebrates, its reduced interactions with co-regulators were not significant in vivo as determined by retinal phenotype analysis in single-copy Nr2e1-null mice carrying the variant R274G.ConclusionsWe showed that MARCoNI is a specific assay to test interactions of NR2E1 with candidate co-regulators. In this way, we unveiled 19 potential co-regulator partners for NR2E1, including eight co-activators. All the candidates here identified need to be further validated using in vitro and in vivo models. This assay was sensitive to point mutations in NR2E1 ligand binding domain making it useful to identify mutations and/or small molecules that alter binding of NR2E1 to protein partners.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3173-5) contains supplementary material, which is available to authorized users.
RESUMO -O objetivo foi avaliar os efeitos da amamentação parcelada nas primeiras seis horas após o nascimento, sobre o peso ao desmame, aos 60 dias de idade e a dosagem de imunoglobulinas G séricas em leitões de porcas de diversas ordens de parto. Trinta e três fêmeas suínas foram usadas em delineamento inteiramente casualizado e em esquema fatorial 2 × 3, composto de dois manejos de leitões pós-nascimento e três agrupamentos por ordem de parto. No manejo tradicional, os leitões disputavam as tetas e mamavam aleatoriamente, enquanto no manejo com alimentação parcelada foram distribuídos em dois grupos para mamar. A coleta de sangue dos leitões foi realizada seis horas após o término do parto para titulação de IgG. Aos 21 e 60 dias de idade, os leitões foram pesados. O manejo de alimentação parcelada não afetou o peso nem a dosagem de IgG sérica dos leitões, independentemente da ordem de parição das porcas, no entanto os leitões das porcas primíparas apresentaram menor peso ao desmame. O manejo de alimentação parcelada resultou em aumento da concentração de IgG sérica dos leitões de porcas primíparas. A amamentação parcelada deve ser utilizada em primíparas, mas não necessariamente em porcas de segunda parição em diante, pois os leitões de primíparas submetidos ao manejo de alimentação parcelada apresentam maiores concentrações de IgG sérica.Palavras chave: IgG, imunidade passiva, manejo de leitões ao parto, suínos Split suckling in relation to the parturition order of sows ABSTRACT -The objective was evaluate the effects of the split suckling at the six first hours after the birth, on the weight at weaning and at 60 days of age and on the serum IgG rate, in piglets from sows of different parturition order. Thirtythree sows were used in a completely randomized design and 2 × 3 factorial arrangement: two post-birth management practices and three groups by parturition order. In the traditional management, piglets fought for the teats and suckled randomly, while in the split suckling, piglets were divided in two groups for suckling. Bood collection was performed at six hours after the end of the birth, for the IgG titration. At 21 and 60 days of age, piglets were weighed. The split suckling management did not affect the weight or the serum IgG rate of the piglets from sows of different parturition order, but the piglets of primiparous sows had lower weaning weight. The split suckling management resulted in increase of serum IgG concentration in piglets of primiparous sows. The split suckling must be used in primiparous sows, but not necessarily in sows of second parturition on, because the piglets of primiparous submitted to the split suckling management present higher concentrations of serum IgG. IntroduçãoOs leitões nascem sem proteção, mas adquirem imunidade passiva por meio da ingestão do colostro nas primeiras horas após o nascimento (Le Dividich et al., 2005;Svendsen et al., 2005). Leitões que não são amamentados por um período de até 60 minutos ou mais após o nascimento (Beynon, 2009) ou que não conseguem in...
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