Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Pups born to mothers fed high folate diet had short-term memory impairment and reduced hippocampal area. ( A ) Three - week-old male mice were tested using the NOR. The discrimination index is the ratio of time spent with the novel object versus time with the familiar object; a negative discrimination index indicates short-term memory impairment. Data from two independent experiments are shown ( n = 15–20/group). ( B ) The average area of the hippocampus and ( C ) the average thickness of the dentate gyrus (DG) were reduced in FASD pups ( n = 5/group). Representative images of hippocampus and DG from CD pup ( D and F , respectively) and FASD pup ( E and G , respectively) are shown. Arrows in the DG indicate where the thickness measurements were performed. Values are means ± SEM; * P < 0.05, *** P < 0.005.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques:

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Pups born to mothers fed high folate diet had pseudo-MTHFR deficiency in liver. ( A ) Total immunoreactive MTHFR protein was reduced in livers of FASD pups. ( B ) The ratio of the phosphorylated isoform (p-MTHFR, less active) to the non-phosphorylated isoform (MTHFR) was increased in FASD pups. ( C ) Representative Western blot of MTHFR in offspring liver. Values are means ± SEM from 5 mice/group; * P <0.05, *** P <0.005.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Western Blot

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Pups born to mothers fed high folate diet had disturbances in choline and acetylcholine metabolism. ( A ) The concentration of phosphocholine (PCho), the storage form of choline, was significantly decreased in liver ( n = 6/group). ( B ) In hippocampus, there was a trend towards reduced glycerophosphocholine (GPC) in FASD pups ( n = 7/group, # P = 0.06). ( C ) mRNA expression of acetylcholinesterase ( AchE) was reduced in cortex of FASD pups ( n = 7/group). ( D ) mRNA levels of the nicotinic receptor of acetylcholine, Chrna7 and AchE in cortex were significantly positively correlated ( n = 7/group; r = 0.894, P < 0.05). ChAT staining to measure ( E ) staining intensity and ( F ) numbers of cholinergic neurons in the Substantia Innominata (SI). There was a trend towards reduced intensity of ChAT staining in the SI of FASD pups ( n = 5/group, # P = 0.07) but no significant difference in number of cholinergic neurons. Values are means ± SEM; * P < 0.05.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Concentration Assay, Expressing, Staining

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Pups of FASD mothers had reduced expression of Dnmt3a mRNA in cortex and hippocampus. There was a significant decrease of Dnmt3a mRNA due to FASD in ( A ) cortex ( n = 7/group) and ( B ) hippocampus of FASD pups ( n = 5/group). ( C ) Dnmt1 and ( D ) Dnmt3b mRNA were not changed by diet in cortex. Values are means ± SEM; * P < 0.05.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Expressing

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: High folic acid intake altered folate metabolism in liver of lactating mothers and led to changes in choline metabolism. ( A ) Total MTHFR protein was reduced in FASD mothers ( n = 5/group). ( B ) The ratio of the phosphorylated MTHFR isoform (less active) to the non-phosphorylated isoform was increased in the FASD group ( n = 5/group). mRNA expression of ( C ) methionine synthase ( Mtr ) was reduced, ( D ) methionine synthase reductase ( Mtrr) was increased and ( E ) Pemt was increased due to FASD ( n = 7/group). ( F ) There was a trend towards lower GPC due to diet in maternal plasma, ( n = 7/group, # P = 0.06). Values are means ± SEM; * P < 0.05, *** P < 0.005.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Expressing, Clinical Proteomics

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Embryos and placentas from FASD mothers were smaller at E17.5. Weights of embryos and placentas were measured at sacrifice. ( A ) FASD led to lower mean body weights/litter of embryos. ( B ) There was a trend toward lower placental weights/litter (# P = 0.08). Values are means ± SEM of 6 l/group; * P < 0.05.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques:

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Maternal FASD altered expression of one-carbon enzymes in liver of E17.5 embryos. ( A ) Total MTHFR protein and ( B ) Mthfr mRNA was reduced in liver of E17.5 FASD embryos. ( C ) There was a trend towards reduced Bhmt mRNA in embryonic liver (# P = 0.08). ( D ) Mat1a and ( E ) Pemt mRNA were both significantly decreased due to diet in embryonic liver. Values are means ± SEM of 7–8 embryos/group; * P < 0.05, *** P < 0.005.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Expressing

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Choline metabolites decreased in E17.5 embryonic liver of FASD mothers. ( A ) Betaine concentration was significantly decreased. ( B ) There was a trend towards reduced choline (# P = 0.06) ( C ) Pcho concentration was significantly decreased. Values are means ± SEM of 6 embryos/group; * P < 0.05, *** P < 0.005.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Concentration Assay

Journal: Human Molecular Genetics

Article Title: High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring

doi: 10.1093/hmg/ddx004

Figure Lengend Snippet: Altered brain development in embryos of FASD mothers. Coronal sections of embryonic brain were stained with H & E. Representative images from embryos in the CD ( A and C ) and FASD ( B and D ) groups are depicted. Images show the cortical layers, the intermediate zone (IZ) and subventricular zone (SVZ). Cortical layers from FASD embryos appeared to be more organized and developed.

Article Snippet: At weaning, wild-type C57BL/6 female mice were placed on amino acid defined Control Diet (CD) (2 mg folic acid/kg diet, recommended level for rodents ( ) (TD.01369, Harlan Laboratories, Inc., Madison, WI, USA) or Folic Acid Supplemented Diet (FASD) which contained 10 times the amount of folic acid (20 mg folic acid/kg diet, TD.09258, Harlan Laboratories, Inc., Madison, WI, USA).

Techniques: Staining