Long-Lasting Effect of Perinatal Exposure to L-tryptophan on Circadian Clock of Primary Cell Lines Established from Male Offspring Born from Mothers Fed on Dietary Protein RestrictionReportar como inadecuado




Long-Lasting Effect of Perinatal Exposure to L-tryptophan on Circadian Clock of Primary Cell Lines Established from Male Offspring Born from Mothers Fed on Dietary Protein Restriction - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Background and Aims

Maternal undernutrition programs metabolic adaptations which are ultimately detrimental to adult. L-tryptophan supplementation was given to manipulate the long-term sequelae of early-life programming by undernutrition and explore whether cultured cells retain circadian clock dysregulation.

Methods

Male rat pups from mothers fed on low protein 8%, LP or control 18%, CP diet were given, one hour before light off, an oral bolus of L-tryptophan 125 mg-kg between Day-12 and Day-21 of age. Body weight, food intake, blood glucose along with the capacity of colonization of primary cells from biopsies were measured during the young 45–55 days and adult 110–130 days phases. Circadian clock oscillations were re-induced by a serum shock over 30 hours on near-confluent cell monolayers to follow PERIOD1 and CLOCK proteins by Fluorescent Linked ImmunoSorbent Assay FLISA and period1 and bmal1 mRNA by RT-PCR. Cell survival in amino acid-free conditions were used to measure circadian expression of MAP-LC3B, MAP-LC3B-FP and Survivin.

Results

Tryptophan supplementation did not alter body weight gain nor feeding pattern. By three-way ANOVA of blood glucose, sampling time was found significant during all phases. A significant interaction between daily bolus Tryptophan, saline and diets LP, CP were found during young p = 0.0291 and adult p = 0.0285 phases. In adult phase, the capacity of colonization at seeding of primary cells was twice lower for LP rats. By three-way ANOVA of PERIOD1 perinuclear-nuclear immunoreactivity during young phase, we found a significant effect of diets p = 0.049, daily bolus p<0.0001 and synchronizer hours p = 0.0002. All factors were significantly interacting p = 0.0148. MAP-LC3B, MAP-LC3B-FP and Survivin were altered according to diets in young phase.

Conclusions

Sequelae of early-life undernutrition and the effects of L-tryptophan supplementation can be monitored non-invasively by circadian sampling of blood D-glucose and on the expression of PERIOD1 protein in established primary cell lines.



Autor: Elizabeth Nascimento, Omar Guzman-Quevedo, Nellie Delacourt, Raquel da Silva Aragão, Georgina Perez-Garcia, Sandra Lopes de Souz

Fuente: http://plos.srce.hr/



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