title: 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.
abstract: OBJECTIVE
  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.
authors: Departamento de Nutrição, Centro de Ciências da Saude, Universidade Federal de Pernambuco, Recife, Pernambuco, Brazil.
authors: Elizabeth
authors: E
authors: Nascimento
authors: Omar
authors: O
authors: Guzman-Quevedo
authors: Nellie
authors: N
authors: Delacourt
authors: Raquel
authors: R
authors: da Silva Aragão
authors: Georgina
authors: G
authors: Perez-Garcia
authors: Sandra Lopes
authors: SL
authors: de Souza
authors: Raul
authors: R
authors: Manhães-de-Castro
authors: Francisco
authors: F
authors: Bolaños-Jiménez
authors: Bertrand
authors: B
authors: Kaeffer
mesh_terms: D000375:Aging / Q000097:blood / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001343:Autophagy / Q000187:drug effects; D015415:Biomarkers / Q000378:metabolism; D001786:Blood Glucose / Q000187:drug effects / Q000378:metabolism; D056926:CLOCK Proteins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D057906:Circadian Clocks / Q000187:drug effects*; D003114:Colony-Forming Units Assay; D018753:Diet, Protein-Restricted; D004734:Energy Metabolism / Q000187:drug effects; D005247:Feeding Behavior / Q000187:drug effects*; D005260:Female; D050152:Intra-Abdominal Fat / Q000033:anatomy & histology / Q000187:drug effects; D007774:Lactation / Q000187:drug effects; D008297:Male; D018811:Maternal Exposure; D010641:Phenotype; D011247:Pregnancy; D051381:Rats; D044967:Serum / Q000378:metabolism; D014364:Tryptophan / Q000494:pharmacology*; D014365:Tryptophan Hydroxylase / Q000378:metabolism; D015430:Weight Gain / Q000187:drug effects
chemical_list: D015415:Biomarkers; D001786:Blood Glucose; D014364:Tryptophan; D014365:Tryptophan Hydroxylase; D056926:CLOCK Proteins