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Frontiers in Biology

ISSN 1674-7984

ISSN 1674-7992(Online)

CN 11-5892/Q

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Front. Biol.    2017, Vol. 12 Issue (4) : 290-297    https://doi.org/10.1007/s11515-017-1457-z
RESEARCH ARTICLE
Thiamine deficiency perturbed energy metabolism enzymes in brain mitochondrial fraction of Swiss mice
Anupama Sharma1(), Renu Bist1, Surender Singh2
1. Department of Bioscience and Biotechnology, Banasthali University, Banasthali, Rajasthan, India
2. Animal Biotechnology Centre, National Dairy Research Institute, Karnal, India
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Abstract

BACKGROUND: Thiamine is an essential cofactor associated with several enzymes in energy metabolism and its deficiency may lead to neurological deficits. Current research evaluated the biochemical and molecular changes in TCA cycle enzymes using the mitochondrial fraction of the brain following thiamine deficiency (TD) in mice.

METHODS: The investigation was carried out on Swiss mice (6-8 week old) allocated into three groups. First group was control; second and third group were made thiamine deficient for 8 and 10 days.

RESULTS: Current study showed that alpha-ketoglutarate dehydrogenase (KGDHC) (thiamine-dependent enzyme) level found to be significantly reduced in experimental groups as compared to control group. In comparison to control group, a significant decrease in the succinate dehydrogenase (SDH) activity was calculated in group II and group III (p<0.0001) mice. Diminished enzymatic activity of fumarase and MDH enzyme in thiamine deficient groups exposed for 8 and 10 days was calculated as compared to control group. The expression analysis of different genes governing TCA cycle enzymes in different experimental groups showed that there was a negotiable change in the expression of fumarase and DLD (dihydrolipoyl dehydrogenase- E3 subunit of KGDHC) whereas a declined in the expression of SDH and two subunits of KGDHC i.e. OGDH (2-oxoglutarate dehydrogenase- E1 subunit of KGDHC) and DLST (dihydrolipoyllysine-residue succinyltransferase- E2 subunit of KGDHC) was observed as compared to control group.

CONCLUSIONS: Hence, current findings strongly entail that TD promotes alteration in energy metabolism in brain mitochondria which will decline the neuronal progression which may lead to neurodegenerative diseases such as Alzheimer’s diseases.
Keywords thiamine      brain      TCA cycle enzymes      m itochondrial dysfunction     
Corresponding Author(s): Anupama Sharma   
Online First Date: 22 August 2017    Issue Date: 13 September 2017
 Cite this article:   
Anupama Sharma,Renu Bist,Surender Singh. Thiamine deficiency perturbed energy metabolism enzymes in brain mitochondrial fraction of Swiss mice[J]. Front. Biol., 2017, 12(4): 290-297.
 URL:  
https://academic.hep.com.cn/fib/EN/10.1007/s11515-017-1457-z
https://academic.hep.com.cn/fib/EN/Y2017/V12/I4/290
Enzyme Subunit of enzymes Forward primer Reverse primer Annealing temperature (°C) Amplicon size
KGDHC OGDH 5′TGCTAAGCCATCTTGCCAGAC 3′ 5′CAACCTAACGTCGACAAACTCG 3′ 51 ~6400bps
DLST 5′AATGGTGTAGATGTAGTAGCAG 3′ 5′CAGCGTTTGCAGAGTCTG 3′ 52 ~2400bps
DLD 5′GCTGACGTGACAGTGATAG 3′ 5′TTTGGTGTCTTCATTCCC 3′ 53 ~2000bps
SDH 5′GAGAACAAGAAGGCATCAGCT 3′ 5′TCAAGGAAGTCAGGGCATGA 3′ 51 ~5400bps
Fumarase 5′ATCTGACGTATTAGGGGGC 3′ 5′AACTGCTCTGCTGTGAGATAG 3′ 53 ~1500bps
MDH 5′GTCGTTGGAGTCACTCGTCTT 3′ 5′TTGGAAGAGATGCTGATGCT 3′ 54 ~5000bps
Tab.1  Primer sequences of different TCA cycle enzymes
Fig.1  KGDHC activity in brain mitochondria of different experimental groups. Results were expressed as mean±SE. a-compared to group I; b-compared to group II; c-compared to group III. *p<0.05, ***p<0.001, ****p<0.0001.
Fig.2  SDH activity in brain mitochondria of different experimental groups. Results were expressed as mean±SE. a-compared to group I; b-compared to group II; c-compared to group III. ***p<0.001, ****p<0.0001.
Fig.3  Fumarase activity in brain mitochondria of different experimental groups. Results were expressed as mean±SE. a-compared to group I; b-compared to group II; c-compared to group III. *p<0.05, ****p<0.0001.
Fig.4  MDH activity in brain mitochondria of different experimental groups. Results were expressed as mean±SE. a-compared to group I; b-compared to group II; c-compared to group III. ****p<0.0001.
Fig.5  RNA of different experimental groups. A- Group I; B- Group II; C-Group III.
Fig.6  (A) Group I- Gene expression of different enzymes in control group. Lane A- MDH, B- Fumarase, C- SDH, D- OGDH, E- DLST, F- DLD and M- marker. (B) Group II- Gene expression of different enzymes in thiamine deficient group for 08 days. Lane A- MDH, B- Fumarase, C- SDH, D- OGDH, E- DLST, F- DLD and M- marker. (C) Group III- Gene expression of different enzymes in thiamine deficient group for 10 days. Lane A- MDH, B- Fumarase, C- SDH, D- OGDH, E- DLST, F- DLD and M- marker.
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