Relation between the intake of milk fat and the occurrence of conjugated linoleic acid in human adipose tissue

Phosphoproteomics is a cutting-edge technique that can be utilized to explore adipose tissue (AT) metabolism by quantifying the repertoire of phospho-peptides (PP) in AT. Dairy cows were supplemented with conjugated linoleic acid (CLA, n = 5) or a control diet (CON, n = 5) from 63 d prepartum to 63 d postpartum; cows were slaughtered at 63 d postpartum and AT was collected. We performed a quantitative phosphoproteomics analysis of subcutaneous (SC) and omental (OM) AT using nanoUPLC-MS/MS and examined the effects of CLA supplementation on the change in the phosphoproteome. A total of 5919 PP were detected in AT, and the abundance of 854 (14.4%) were differential between CON and CLA AT (p ≤ 0.05 and fold change ± 1.5). The abundance of 470 PP (7.9%) differed between OM and SC AT, and the interaction treatment vs. AT depot was significant for 205 PP (3.5% of total PP). The integrated phosphoproteome demonstrated the up- and downregulation of PP from proteins related to lipolysis and lipogenesis, and phosphorylation events in multiple pathways, including the regulation of lipolysis in adipocytes, mTOR signaling, insulin signaling, AMPK signaling, and glycolysis. The differential regulation of phosphosite on a serine residue (S777) of fatty acid synthase (FASN) in AT of CLA-supplemented cows was related to lipogenesis and with more phosphorylation sites compared to acetyl-coenzyme A synthetase (ACSS2). Increased protein phosphorylation was seen in acetyl-CoA carboxylase 1 (ACACA;8 PP), FASN (9 PP), hormone sensitive lipase (LIPE;6 PP), perilipin (PLIN;3 PP), and diacylglycerol lipase alpha (DAGLA;1 PP) in CLA vs. CON AT. The relative gene expression in the SC and OM AT revealed an increase in LIPE and FASN in CLA compared to CON AT. In addition, the expression of DAGLA, which is a lipid metabolism enzyme related to the endocannabinoid system, was 1.6-fold higher in CLA vs. CON AT, and the expression of the cannabinoid receptor CNR1 was reduced in CLA vs. CON AT. Immunoblots of SC and OM AT showed an increased abundance of FASN and a lower abundance of CB1 in CLA vs. CON. This study presents a complete map of the SC and the OM AT phosphoproteome in dairy cows following CLA supplementation and discloses many unknown phosphorylation sites, suggestive of increased lipid turnover in AT, for further functional investigation.

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Influence of Dietary Fish Oil on Conjugated Linoleic Acid and Other Fatty Acids in Milk Fat from Lactating Dairy Cows

Journal of Dairy Science ◽

10.3168/jds.s0022-0302(00)75155-1 ◽

2000 ◽

Vol 83 (11) ◽

pp. 2620-2628 ◽

Cited By ~ 139

Author(s):

D.C. Donovan ◽

D.J. Schingoethe ◽

R.J. Baer ◽

J. Ryali ◽

A.R. Hippen ◽

...

Keyword(s):

Fatty Acids ◽

Linoleic Acid ◽

Fish Oil ◽

Dairy Cows ◽

Conjugated Linoleic Acid ◽

Milk Fat ◽

Lactating Dairy Cows ◽

Dietary Fish

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A new conjugated linoleic acid isomer, 7 trans, 9 cis-octadecadienoic acid, in cow milk, cheese, beef and human milk and adipose tissue

Lipids ◽

10.1007/s11745-998-0273-z ◽

1998 ◽

Vol 33 (8) ◽

pp. 803-809 ◽

Cited By ~ 115

Author(s):

Martin P. Yurawecz ◽

John A. G. Roach ◽

Najibullah Sehat ◽

Magdi M. Mossoba ◽

John K. G. Kramer ◽

...

Keyword(s):

Adipose Tissue ◽

Linoleic Acid ◽

Human Milk ◽

Conjugated Linoleic Acid ◽

Octadecadienoic Acid ◽

Cow Milk ◽

Conjugated Linoleic Acid Isomer ◽

Linoleic Acid Isomer

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A Supplement Containing Trans-10, Cis-12 Conjugated Linoleic Acid Reduces Milk Fat Yield but Does Not Alter Organ Weight or Body Fat Deposition in Lactating Ewes

Journal of Nutrition ◽

10.3945/jn.110.126490 ◽

2010 ◽

Vol 140 (11) ◽

pp. 1949-1955 ◽

Cited By ~ 17

Author(s):

Liam A. Sinclair ◽

Weerasinghe M. P. B. Weerasinghe ◽

Robert G. Wilkinson ◽

Michael J. de Veth ◽

Dale E. Bauman

Keyword(s):

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Body Fat ◽

Milk Fat ◽

Fat Deposition ◽

Organ Weight ◽

Fat Yield

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Seasonal Variation in Conjugated Linoleic Acid and Vaccenic Acid in Milk Fat of Sheep and its Transfer to Cheese and Ricotta

Journal of Dairy Science ◽

10.3168/jds.s0022-0302(05)72797-1 ◽

2005 ◽

Vol 88 (4) ◽

pp. 1311-1319 ◽

Cited By ~ 119

Author(s):

A. Nudda ◽

M.A. McGuire ◽

G. Battacone ◽

G. Pulina

Keyword(s):

Seasonal Variation ◽

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Milk Fat ◽

Vaccenic Acid

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The effect of trans- 10, cis- 12 conjugated linoleic acid on milk fat synthesis in lactating sheep

Proceedings of the British Society of Animal Science ◽

10.1017/s1752756200010036 ◽

2005 ◽

Vol 2005 ◽

pp. 92-92 ◽

Cited By ~ 1

Author(s):

L. A. Sinclair ◽

A. L. Lock ◽

J. W. Perfield ◽

B. M. Teles ◽

D. E. Bauman

Keyword(s):

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Milk Fat ◽

Milk Protein ◽

Potent Inhibitor ◽

Curvilinear Relationship ◽

Fat Synthesis ◽

Fat Yield ◽

Protein Output ◽

Milk Fat Synthesis

Trans- 10, cis- 12 conjugated linoleic acid (CLA), a biohydrogenation intermediate produced in the rumen, is a potent inhibitor of milk fat synthesis. Data from a number of studies where various doses of trans -10, cis -12 CLA have been abomasally infused demonstrate a curvilinear relationship between the percent reduction in milk fat yield and both the dose of trans- 10, cis- 12 CLA infused and the milk fat content of trans- 10, cis- 12 CLA. In addition to a reduction in milk fat output, under some circumstances an increase in milk yield and milk protein output are observed. To date, there has been no examination of the effects of trans- 10, cis- 12 CLA on milk fat synthesis in lactating sheep. The current study was therefore designed to determine if trans- 10, cis- 12 CLA would inhibit milk fat synthesis in lactating sheep. In order to test the effectiveness of trans- 10, cis- 12 CLA in inhibiting milk fat synthesis we used a lipid-encapsulated trans- 10, cis- 12 CLA supplement (LE-CLA) as a means to provide the trans- 10, cis- 12 CLA isomer post-ruminally.

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Isomerisation of cis-9 trans-11 conjugated linoleic acid (CLA) to trans-9 trans-11 CLA during acidic methylation can be avoided by a rapid base catalysed methylation of milk fat

Journal of Dairy Research ◽

10.1017/s0022029908003518 ◽

2008 ◽

Vol 75 (3) ◽

pp. 354-356 ◽

Cited By ~ 18

Author(s):

Michael R F Lee ◽

John K S Tweed

Keyword(s):

Fatty Acid ◽

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Milk Fat ◽

Fatty Acid Methyl Esters ◽

Methyl Esters ◽

The Other ◽

Gas Liquid Chromatography ◽

Freeze Dried ◽

Acidic Conditions

This study investigated the evolution of trans-9 trans-11 conjugated linoleic acid (CLA) from cis-9 trans-11 CLA during methylation and its avoidance through a rapid base methylation of milk fat. The study examined three conditions shown to result in loss of cis-9 trans-11 CLA during methylation namely: temperature, methylation time, water contamination in old reagents and acidic conditions. Three techniques currently used for the conversion of milk fat into fatty acid methyl esters for analysis of CLA content by gas liquid chromatography and a fourth procedure designed to eliminate acidic conditions and to limit methylation temperature and time were used. The four methods were: (i) acidic methylation (AM); (ii) acidic and basic bimethylation with fresh reagents (FBM); (iii) acidic and basic bimethylation with pre-prepared reagents (PBM) and (iv) basic methylation (BM). Each regime was carried out on six milk samples over two periods and methylated 1 ml freeze-dried milk (n=12 per regime). Total CLA was not different across methylation regimes (0·30 mg/ml). Isomer cis-9 trans-11 was higher (P<0·01) with BM than the other regimes and lowest with AM: 21·2, 17·8, 18·8 and 14·7 mg/100 ml for BM, FBM, PBM and AM, respectively. The inverse relationship was shown for trans-9 trans-11 with higher (P<0·001) amounts with AM than the other regimes and lowest with BM: 0·57, 2·55, 2·36 and 3·69 mg/100 ml for BM, FBM, PBM and AM, respectively. The trans-10 cis-12 isomer was also shown to alter with methylation procedure being higher (P<0·001) with AM than the other regimes: 0·43, 0·47, 0·29 and 1·20 mg/100 ml for BM, FBM, PBM and AM, respectively. Validation with known CLA free fatty acid and triacylglycerol standards confirmed that AM resulted in conversion of cis-9 trans-11 to trans-9 trans-11, and also elevated trans-10 cis-12 whilst BM of triacylglycerol CLA did not isomerise cis-9 trans-11 and was comparable to FBM.

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