Topologically associating domains in the POLLED region are the same for Angus- and Brahman-specific Hi-C reads from F1 hybrid fetal tissue.

Horns, a form of headgear carried by Bovidae, have ethical and economic implications for ruminant production species such as cattle and goats. Hornless (polled) individuals are preferred. In cattle, four genetic variants (Celtic, Friesian, Mongolian and Guarani) are associated with the polled phenotype, which are clustered in a 300-kb region on chromosome 1. As the variants are intergenic, the functional effect is unknown. The aim of this study was to determine if the POLLED variants affect chromatin structure or disrupt enhancers using publicly available data. Topologically associating domains (TADs) were analyzed using Angus- and Brahman-specific Hi-C reads from lung tissue of an Angus (Celtic allele) cross Brahman (horned) fetus. Predicted bovine enhancers and chromatin immunoprecipitation sequencing peaks for histone modifications associated with enhancers (H3K27ac and H3K4me1) were mapped to the POLLED region. TADs analyzed from Angus- and Brahman-specific Hi-C reads were the same, therefore, the Celtic variant does not appear to affect this level of chromatin structure. The Celtic variant is located in a different TAD from the Friesian, Mongolian, and Guarani variants. Predicted enhancers and histone modifications overlapped with the Guarani and Friesian variants but not the Celtic or Mongolian variants. This study provides insight into the mechanisms of the POLLED variants for disrupting horn development. These results should be validated using data produced from the horn bud region of horned and polled bovine fetuses.

Understanding the effects of the bovine POLLED variants.

Horns are paired appendages on the head of bovine species, comprising an inner bony core and outer keratin sheath. The horn bud forms during early fetal development but ossification of the developing horn does not occur until approximately 1 month after birth. Little is known about the genetic pathways that lead to horn growth. Hornless, or polled, animals are found in all domestic bovids. Histological studies of bovine fetuses have shown that the horn bud does not form in polled individuals. There are currently four known genetic variants for polledness in cattle on BTA1. All of the variants are intergenic, but probably affect regulation of nearby genes or long non-coding RNAs. Transcriptomic studies suggest that the expression of two nearby long non-coding RNAs are affected by the Celtic POLLED variant, but further studies are required to confirm these data. Candidate genes located elsewhere in the genome are involved in regulating bone formation and epithelial-to-mesenchymal transition. Expression of one of these candidate genes, RXFP2, appears to be reduced in the fetal horn bud of polled animals carrying the Celtic variant compared with horned individuals. Investigating horn ontogenesis and the genetic pathway by which the POLLED variants prevent horn development has implications for cattle breeding. If the genetic basis of horn bud formation and polledness is better understood, then new targets may be identified for precision genome editing to create polled individuals.

Vitamin A and marbling attributes: Intramuscular fat hyperplasia effects in cattle.

Twenty Angus steers were fed a diet low in ß-carotene and vitamin A for 10months. Ten steers were supplemented with vitamin A weekly, while the other ten steers did not receive any additional vitamin A. The results demonstrated that the restriction of vitamin A intake increased intramuscular fat (IMF) by 46%. This was a function of the total number of marbling flecks increasing by 22% and the average marbling fleck size increasing by 14%. Vitamin A restriction resulted in marbling flecks that were less branched (22%) and slightly more round (4%) with an increased minor axis length (7%). However, restricting vitamin A did not affect the size of the intramuscular or subcutaneous adipocyte cells or the subcutaneous fat depth. The results suggest that vitamin A affects the amount of marbling and other attributes of the marbling flecks due to hyperplasia rather than hypertrophy. This may explain why vitamin A restriction specifically affects IMF rather than subcutaneous fat deposition.

Modeling tenderness for genetic and quantitative trait loci analyses.

Tenderness is the one of the most important attributes of beef with regards to the intent of consumers to repurchase. Beef tenderness is frequently measured objectively as Warner-Bratzler shear force. However, to better understand the genetics underlying tenderness, better descriptors were derived herein. Two traits, adjusted shear force and aging rate, were calculated to incorporate measures of Warner-Bratzler shear force repeated over aging time of 2 muscles, longissimus dorsi (LD) and semitendinosus (ST). Analysis of these traits indicated that the shear forces of the 2 muscles are lowly correlated and aging rate, as calculated, was not correlated with adjusted shear force. Breed, SNP, and QTL effects were analyzed for the 4 traits (2 traits × 2 muscles). For adjusted shear force, the QTL were located on cattle chromosomes BTA 5, 18, 25, and 29. With the exception of the QTL on BTA 5, the adjusted shear force QTL were observed for either the LD or the ST but not both muscles. The QTL for aging rate were located on BTA 1, 4, 7, 11, 13, 19, and 20.

Effects of quantitative trait loci and the myostatin locus on trace and macro elements (minerals) in bovine liver, muscle and kidney.

A quantitative trait locus (QTL) study of the concentrations of 14 trace and macro elements (minerals) in tissues of beef cattle was conducted in New Zealand. Back-cross calves with Jersey and Limousin ancestry (202 heifers and 211 steers) were generated using first-cross sires. This paper reports on testing for effects of QTL on the concentrations of minerals in liver, kidney and muscle in cattle at slaughter, following a growth phase during which rearing and finishing stages were on pasture. Fifteen QTL were identified (P < 0.05) on a genome-wide basis in combined-sire and within-sire analyses. In addition, the possible effect of the Limousin myostatin F94L allele was tested by fitting each calf's myostatin genotype, and 16 QTL were identified. Twelve were in common with those QTL identified previously, comprising six affecting the liver (copper and zinc, on two chromosomes each; plus iron and molybdenum), three affecting the kidney (calcium, copper and iron), and three affecting muscle (iron, strontium and zinc).

Genetic mapping of quantitative trait loci for meat quality and muscle metabolic traits in cattle.

A whole-genome scan was carried out in New Zealand and Australia to detect quantitative trait loci (QTL) for live animal and carcass composition traits and meat quality attributes in cattle. Backcross calves (385 heifers and 398 steers) were generated, with Jersey and Limousin backgrounds. The New Zealand cattle were reared and finished on pasture, whilst Australian cattle were reared on grass and finished on grain for at least 180 days. This paper reports on meat quality traits (tenderness measured as shear force at 4-5 ages on two muscles as well as associated traits of meat colour, pH and cooking loss) and a number of metabolic traits. For meat quality traits, 18 significant QTL (P < 0.05), located in nine linkage groups, were detected on a genome-wise basis, in combined-sire (seven QTL) or within-sire analyses (11 QTL). For metabolic traits, 11 significant QTL (P < 0.05), located in eight linkage groups, were detected on a genome-wise basis, in combined-sire (five QTL) or within-sire analyses (six QTL). BTA2 and BTA3 had QTL for both metabolic traits and meat quality traits. Six significant QTL for meat quality and metabolic traits were found at the proximal end of chromosome 2. BTA2 and BTA29 were the most common chromosomes harbouring QTL for meat quality traits; QTL for improved tenderness were associated with Limousin-derived and Jersey-derived alleles on these two chromosomes, respectively.

Quantitative trait loci for organ weights and adipose fat composition in Jersey and Limousin back-cross cattle finished on pasture or feedlot.

A QTL study of live animal and carcass traits in beef cattle was carried out in New Zealand and Australia. Back-cross calves (385 heifers and 398 steers) were generated, with Jersey and Limousin backgrounds. This paper reports on weights of eight organs (heart, liver, lungs, kidneys, spleen, gastro-intestinal tract, fat, and rumen contents) and 12 fat composition traits (fatty acid (FA) percentages, saturated and monounsaturated FA subtotals, and fat melting point). The New Zealand cattle were reared and finished on pasture, whilst Australian cattle were reared on grass and finished on grain for at least 180 days. For organ weights and fat composition traits, 10 and 12 significant QTL locations (P<0.05), respectively, were detected on a genome-wide basis, in combined-sire or within-sire analyses. Seven QTL significant for organ weights were found at the proximal end of chromosome 2. This chromosome carries a variant myostatin allele (F94L), segregating from the Limousin ancestry, and this is a positional candidate for the QTL. Ten significant QTL for fat composition were found on chromosomes 19 and 26. Fatty acid synthase and stearoyl-CoA desaturase (SCD1), respectively, are positional candidate genes for these QTL. Two FA QTL found to be common to sire groups in both populations were for percentages of C14:0 and C14:1 (relative to all FAs) on chromosome 26, near the SCD1 candidate gene.

Genetic variation in the beta, beta-carotene-9', 10'-dioxygenase gene and association with fat colour in bovine adipose tissue and milk.

beta, beta-carotene-9', 10'-dioxygenase (BCO2) plays a role in cleaving beta-carotene eccentrically, and may be involved in the control of adipose and milk colour in cattle. The bovine BCO2 gene was sequenced as a potential candidate gene for a beef fat colour QTL on chromosome (BTA) 15. A single nucleotide base change located in exon 3 causes the substitution of a stop codon (encoded by the A allele) for tryptophan(80) (encoded by the G allele) (c. 240G>A, p.Trp80stop, referred to herein as SNP W80X). Association analysis showed significant differences in subcutaneous fat colour and beta-carotene concentration amongst cattle with different BCO2 genotypes. Animals with the BCO2 AA genotype had more yellow beef fat and a higher beta-carotene concentration in adipose tissues than those with the GA or GG genotype. QTL mapping analysis with the BCO2 SNP W80X fitted as a fixed effect confirmed that this SNP is likely to represent the quantitative trait nucleotide (QTN) for the fat colour-related traits on BTA 15. Moreover, animals with the AA genotype had yellower milk colour and a higher concentration of beta-carotene in the milk.

Quantitative trait loci for live animal and carcass composition traits in Jersey and Limousin back-cross cattle finished on pasture or feedlot.

A quantitative trait locus (QTL) study was carried out in two countries, recording live animal and carcass composition traits. Back-cross calves (385 heifers and 398 steers) were generated, with Jersey and Limousin breed backgrounds. The New Zealand cattle were reared on pasture to carcass weights averaging 229 kg, whilst the Australian cattle were reared on grass and finished on grain (for at least 180 days) to carcass weights averaging 335 kg. From 11 live animal traits and 31 carcass composition traits respectively, 5 and 22 QTL were detected in combined-sire analyses, which were significant (P < 0.05) on a genome-wise basis. Fourteen significant traits for carcass composition QTL were on chromosome 2 and these were traits associated with muscling and fatness. This chromosome carried a variant myostatin allele (F94L), segregating from the Limousin ancestry. Despite very different cattle management systems between the two countries, the two populations had a large number of QTL in common. Of the 18 traits which were common to both countries, and which had significant QTL at the genome-wise level, eight were significant in both countries.

Limousin myostatin F94L variant affects semitendinosus tenderness.

Texture parameters (peak force and compression), muscle myofibre diameter, and hydroxyproline were measured in semitendinosus samples from a cattle gene-mapping herd. The data were analysed to determine the relationships between these traits. The traits were also mapped by genetic linkage analysis to identify quantitative trait loci, and hence, candidate genes for these traits. Neither texture parameters were affected by the muscle structural traits of myofibre diameter or collagen content (as measured by hydroxyproline), despite significant variation in these traits between animals. QTL for the texture parameters of peak force and compression, as well as collagen content, were found on cattle chromosome 2 (BTA2) and attributed to the myostatin gene. Within the cattle population used for the QTL mapping, a gene variant of myostatin, F94L, has been previously shown to increase muscle mass, predominantly in the semitendinosus. It was determined herein that the F94L myostatin homozygous animals had more tender meat as measured by both peak force and compression. The variant was also responsible for a reduction in the collagen/elastin content of muscle. The myostatin F94L variant had no effect on muscle myofibre diameter of the semitendinosus, even though the variant causes substantial increases in muscle mass. Consequently, the increase in muscle mass of the variant must be due to myofibre hyperplasia and not hypertrophy. In addition, myostatin effects on tenderness are caused by changes in the extracellular matrix rather than muscle myofibre diameter.

Effects of the myostatin F94L substitution on beef traits.

This study investigated the effects of a SNP in the myostatin gene (MSTN or growth differentiation factor 8, GDF8) on birth, growth, carcass, and beef quality traits in Australia (Aust.) and New Zealand (NZ). The SNP is a cytosine to adenine transversion in exon 1, causing an amino acid substitution of leucine for phenylalanine(94) (F94L). The experiment used crosses between the Jersey and Limousin breeds, with the design being a backcross using first-cross bulls of Jersey x Limousin or Limousin x Jersey breeding, mated to Jersey and Limousin cows. Progeny were genotyped for the myostatin SNP and phenotyped in Aust., with finishing on feedlot (366 calves, over 3 birth years) and in NZ with finishing on pasture (416 calves, over 2 birth years). The effect of the F94L allele (A allele) on birth and growth traits was not significant. The F94L allele in Limousin backcross calves was associated with an increase in meat weight (7.3 and 5.9% of the trait mean in Aust. and NZ, respectively, P < 0.001), and a reduction in fat depth (-13.9 and -18.7% of the trait means on live calves (600 d) and carcasses, respectively, Aust. only, P < 0.001), intramuscular fat content (-8.2% of the trait mean in Aust., P < 0.05; -7.1% in NZ, not significant), total carcass fat weight (-16.5 and -8.1% of the trait mean, Aust. and NZ; P < 0.001 and P < 0.05, respectively). Meat tenderness, pH, and cooking loss of the M. longissimus dorsi were not affected by the F94L variant. In the Jersey backcross calves, additive and dominance effects were confounded because the F94L allele was not segregating in the Jersey dams. The combined effects, however, were significant on LM area (4.4% in both Aust., P < 0.05, and NZ, P < 0.01), channel fat (-11.7%, NZ only, P < 0.01), rib fat depth (-11.2%, NZ only, P < 0.05), and carcass fat weight (-7.1%, NZ only, P < 0.05). The results provide strong evidence that this myostatin F94L variant provides an intermediate and more useful phenotype than the more severe double-muscling phenotype caused by knockout mutations in the myostatin gene.

Effect of myostatin F94L on carcass yield in cattle.

In this study, a highly significant quantitative trait locus (QTL) for meat percentage, eye muscle area (EMA) and silverside percentage was found on cattle chromosome 2 at 0-15 cM, a region containing the positional candidate gene growth differentiation factor 8 (GDF8), which has the common alias myostatin (MSTN). Loss-of-function mutations in the MSTN gene are known to cause an extreme 'double muscling' phenotype in cattle. In this study, highly significant associations of MSTN with cattle carcass traits were found using maternally inherited MSTN haplotypes from outbred Limousin and Jersey cattle in a linkage disequilibrium analysis. A previously reported transversion in MSTN (AF320998.1:g.433C>A), resulting in the amino acid substitution of phenylalanine by leucine at position 94 of the protein sequence (F94L), was the only polymorphism consistently related to increased muscling. Overall, the size of the g.433C>A additive effect on carcass traits was moderately large, with the g.433A allele found to be associated with a 5.5% increase in silverside percentage and EMA and a 2.3% increase in total meat percentage relative to the g.433C allele. The phenotypic effects of the g.433A allele were partially recessive. This study provides strong evidence that a MSTN genotype can produce an intermediate, non-double muscling phenotype, which should be of significant value for beef cattle producers.

Genotypic effects of calpain 1 and calpastatin on the tenderness of cooked M. longissimus dorsi steaks from Jersey x Limousin, Angus and Hereford-cross cattle.

Single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1) and calpastatin (CAST) genes were studied to determine their effects on meat tenderness in Bos taurus cattle. Strip loins (M. longissimus dorsi) were removed from cattle in four resource populations after slaughter (n = 1042), aged under controlled conditions until fixed times after rigor mortis, cooked and measured using a tenderometer. Animals were genotyped for the CAPN1 SNP c.947C>G (p.Ala316Gly; AF252504) and for the CAST SNP c.2959A>G (AF159246). Frequencies of CAPN1 C alleles ranged from 23% to 68%, and CAST A alleles from 84% to 99.5%. From all data combined, the CAPN1 CC genotype (compared with the GG genotype) was associated with a 20.1 +/- 1.7% reduced average shear force at intermediate stages of ageing (P < 0.001) and with a 9.5 +/- 1.3% reduction near ultimate tenderness (P < 0.001). The heterozygote was intermediate. For CAST, corresponding values for AA compared with AG genotypes were reductions of 8.6 +/- 2.0% and 5.1 +/- 1.6% respectively (both P < 0.001), but there were too few GG genotypes for comparison. There were small interactions between the CAPN1 and CAST genotypes. For the CAPN1 and CAST genotypes combined, the maximal genotype effect in average shear force was 25.7 +/- 5.5% (P < 0.001) at intermediate stages and 15.2 +/- 4.8% near ultimate tenderness (P < 0.01).

Effect of low vitamin A status on fat deposition and fatty acid desaturation in beef cattle.

A group of Angus beef cattle was removed from temperate pastures and fed a very low beta-carotene cereal-based ration in a feedlot for over 300 d. Half the group was supplemented weekly with retinyl palmitate (at the rate of 60,000 IU vitamin A/100 live weight (LW)/day), sufficient to offset clinical vitamin A deficiency; the other half received no supplement. Blood was sampled from all animals at biweekly intervals to assess beta-carotene and vitamin A status. Adipose tissue was sampled by biopsy on three occasions throughout the experimental period and at slaughter to assess FA composition. Muscle was sampled at slaughter to determine the intramuscular fat content. The mean plasma concentration of beta-carotene of all animals fell from an initial value of 20.1 to 5.2 microg/mL at 14 d, to 1.4 microg/mL at 35 d, and to zero at 105 d. Mean vitamin A in plasma was not significantly different between the treatment groups initially. The values then rose to almost twice their initial values by 35 d, but subsequently fell to below initial values by day 119. Thereafter, plasma vitamin A of the supplemented group was significantly greater than that of the unsupplemented group (P < 0.05). Muscle samples at slaughter from supplemented animals contained significantly (P < 0.01) more intramuscular lipid (13.0 vs. 9.6%). Major changes occurred over time in FA composition in both groups. Saturated FA decreased as monounsaturated FA increased over the first 60 d. An index of desaturation of FA was significantly lower (P < 0.001) in the vitamin A-supplemented group than in the nonsupplemented group. M.P. of the adipose tissue of nonsupplemented animals was 32.3 degrees C, significantly less (P< 0.05) than that of supplemented animals (34.1 degrees C). Feeding vitamin A was associated with less intramuscular fat but with a less desirable (less unsaturated, more solid) FA profile.

Molecular cloning and characterization of bovine PRKAG3 gene: structure, expression and single nucleotide polymorphism detection.

The protein kinase adenosine monophosphate-activated gamma3-subunit (PRKAG3) gene encodes a muscle-specific isoform of the regulatory gamma-subunit of adenosine monophosphate-activated protein kinase, which plays a key role in regulating energy homeostasis in eucaryotes. It is well known that mutations in the PRKAG3 gene affect high glycogen content in the porcine skeletal muscle and, consequently, meat quality. The genomic structure and sequence of the bovine PRKAG3 were analysed from a Korean cattle BAC clone. The bovine PRKAG3 gene comprises 13 exons and spans approximately 6.8 kb on BTA2. From 5' and 3'-rapid amplification of cDNA ends experiments, the full-length cDNA of bovine PRKAG3 has been identified, encoding a deduced protein of 465 amino acids. Two splice isoforms, generated by the alternative splicing of exon 2, were also identified. Northern blot analysis demonstrated that, similar to other species, the bovine PRKAG3 transcript was only expressed in skeletal muscle. Seven single nucleotide polymorphisms, including two previously identified variants, were detected in four Bos taurus cattle breeds. The bovine PRKAG3 gene described in this study may be involved in muscle-related genetic diseases or meat quality traits in cattle.

Differences in delta9 desaturase activity between Jersey- and Limousin-sired cattle.

An experiment examined delta9 desaturase activity and FA composition in subcutaneous adipose tissue in two differing breeds of cattle. Jersey-sired cattle had significantly higher rates of desaturase activity than Limousin-sired cattle (1.55 vs. 0.75 nmol/mg protein/min). This difference was also demonstrated by a lower concentration of individual (e.g., 18:0) and total saturated FA (38.3 vs. 45.1 wt%), and a higher concentration of individual (e.g., 16:1) and total monounsaturated FA (58.2 vs. 52.7 wt%) in the Jersey animals. Other indices of desaturation calculated from the FA composition showed this same difference. The slip point of adipose tissue of Jersey cattle (36.8 degrees C) was significantly lower than that of Limousin cattle (39.2 degrees C), but Jersey adipose tissue had a greater content of beta-carotene. The positive relationship between adipose tissue beta-carotene and desaturation opposes the negative relationship between dietary beta-carotene and desaturation determined elsewhere. These results, however, lead to the hypothesis that some cattle have a reduced capacity to metabolize beta-carotene to various forms of vitamin A, a compound that can reduce delta9 desaturase enzyme activity. In addition, the higher level of intramuscular fat in Jersey cattle (6.97 vs. 3.82%) is possibly related to a lack of inhibition of the adipocyte differentiation genes by vitamin A.

Genetic variation in fatness and fatty acid composition of crossbred cattle.

Mature Hereford cows (766) were mated to 97 sires from seven breeds (Jersey, Wagyu, Angus, Hereford, South Devon, Limousin, and Belgian Blue), resulting in 1,215 calves born over 4 yr (1994 to 1997). These cattle comprised Australia's 'Southern Crossbreeding Project." Heifers were slaughtered at an average of 16 mo with hot standard carcass weight of 219 kg and 9 mm fat over the rump. Steers were slaughtered at an average of 23 mo with carcass weight of 319 kg and 13 mm fat over the rump. Meat and fat samples were taken from the carcass on the day after slaughter for subsequent laboratory analysis of i.m. fat content and fatty acid composition. Data were analyzed using uni- and bivariate animal models containing fixed effects of cohort, management group, birth month, and sire breed. March-born calves had fat with a 0.5 degrees C lower melting point, 0.6% higher total monounsaturated fatty acids, and 0.7% higher fatty acid desaturation index than calves born in April. Steers born in 1997 were the only cohort finished on pasture, and they had much more yellow fat than the other cohorts. Four heavy breed crosses (Angus, South Devon, Limousin, and Belgian Blue) averaged 284 kg carcass weight, followed by purebred Hereford (268 kg), Wagyu (244 kg) and Jersey (236 kg). Angus had the greatest fat depth (14.3 mm), ahead of Hereford and Wagyu (11.9 mm), Jersey (10.7 mm), South Devon and Limousin (9.9 mm) and Belgian Blue (8.0 mm). Jersey, Wagyu, and Angus had themost i.m. fat (4.6%), followed by Hereford and South Devon (3.8%), and Limousin and Belgian Blue (3.1%). The highly marbled Jersey and Wagyu had softer fat (6% lower fat melting point) than the other breeds. Angus were more highly marbled, similar to Jersey and Wagyu, but had harder fat similar to the leaner breeds. Heritabilities for all traits were low to moderate (16 to 36%). Genetic correlations between fatty add composition and carcass traits were not significant, indicating little evidence of antagonisms between traits that would prevent genetic progress in both production and quality.

Consensus and comprehensive linkage maps of bovine chromosome 24.

This study describes development of a consensus genetic linkage map of bovine chromosome 24 (BTA24). Eight participating laboratories contributed data for 58 unique markers including a total of 25 409 meioses. Eighteen markers, which were typed in more than one reference population, were used as potential anchors to generate a consensus framework map. The framework map contained 16 loci ordered with odds greater than 1000:1 and spanned 79.3 cM. Remaining markers were included in a comprehensive map relative to these anchors. The resulting BTA24 comprehensive map was 98.3 cM in length. Average marker intervals were 6.1 and 2.5 cM for framework and comprehensive maps, respectively. Marker order was generally consistent with previously reported BTA24 linkage maps. Only one discrepancy was found when comparing the comprehensive map with the published USDA-MARC linkage map. Integration of genetic information from different maps provides a high-resolution BTA24 linkage map.

Green tea upregulates the low-density lipoprotein receptor through the sterol-regulated element binding Protein in HepG2 liver cells.

Green tea from Camellia sinensis lowers plasma cholesterol in animal models of hypercholesterolemia. The aim of this study was to determine the effects of green tea on the expression of the hepatic low-density lipoprotein (LDL) receptor, a cell surface protein involved in the control of plasma cholesterol. Incubating human HepG2 liver cells in culture with green tea increased both LDL receptor binding activity and protein. An ethyl acetate extract of green tea, containing 70% (w/w) catechins, also increased the LDL receptor binding activity, protein, and mRNA, indicating that (1) the effect was at the level of gene transcription and that (2) the catechins were the active constituents. The mechanism by which green tea up-regulated the LDL receptor was then investigated. Green tea decreased the cell cholesterol concentration (-30%) and increased the conversion of the sterol-regulated element binding protein (SREBP-1) from the inactive precursor form to the active transcription-factor form. Consistent with this, the mRNA of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme in cholesterol synthesis, was also increased by green tea. In conclusion, green tea up-regulated the LDL receptor in HepG2 cells. The effect was most likely mediated through SREBP-1 in response to a decrease in the intracellular cholesterol concentration. The LDL receptor may therefore play a role in the hypocholesterolemic effect of green tea in vivo.