Gene 574 (2015) 204209

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Gene
journal homepage: www.elsevier.com/locate/gene

Research paper

Genetic polymorphisms of the AMPD1 gene and their correlations with

IMP contents in Fast Partridge and Lingshan chickens
Jin Hu, Ping Yu, Xiaoling Ding, Minglong Xu, Baoping Guo, Yinxue Xu
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China

a r t i c l e

i n f o

Article history:
Received 28 January 2015
Received in revised form 3 August 2015
Accepted 5 August 2015
Available online 12 August 2015
Keywords:
AMPD1
IMP
SNP
Fast Partridge chicken
Lingshan chicken

a b s t r a c t
The object of this study was to evaluate associations between the adenosine monophosphate deaminase 1
(AMPD1) gene polymorphisms and inosine monophosphate acid (IMP) contents of chicken to provide a molecular marker for breeding. Three single nucleotide polymorphisms (SNPs), g.4064G/A, g.5573A/G and g.6805G/A
were detected in exons IV, VI, and VIII of the AMPD1 gene in Fast Partridge and Lingshan chickens, respectively. All
were purine conversion and caused no alteration in amino acid sequence. Statistical analysis revealed that
Lingshan chicken with the homozygous genotype AA at position 4064 and 6805 had a signicantly greater IMP
content than those with the GG genotype (P b 0.05). Fast Partridge chicken with the genotype GG at position
6805 had a signicantly greater IMP content compared with those with the AA genotype (P b 0.05). In conclusion,
the polymorphism at g.6805A/G was correlated with IMP content (P b 0.05) in both Fast Partridge and Lingshan
chickens. The results in our study suggest that SNP 6805A/G can be used as a possible candidate marker of IMP
content of chicken.
2015 Elsevier B.V. All rights reserved.

1. Introduction
Meat quality is an important economic trait that includes texture,
nutrition value, and avor. However, meat quality is a comprehensive
evaluation characteristic in livestock production. The latest results
show that muscle avor is closely related to the amino acids present
(especially the glutamate and glycine), IMP content, and intramuscular
fat (IMF) content. Among these parameters, IMF content is closely
related to tenderness, shear forces, etc., whereas IMP content is one of
the main contributors to the avor and aroma of muscle (Horio and
Kawamura, 1990; Yamaguchi and Ninomiya, 2000).
Adenosine monophosphate deaminase (AMPD) is a central enzyme
in eukaryotic energy metabolism due to its participation in purine
nucleotide catabolism (Zimmermann, 1992), in which it catalyzes the
hydrolytic deamination of AMP to IMP and an ammonium ion. Its
multiple isoforms have been isolated from various human and animal
tissues and are named for the source tissue from which they were puried (Ogasawara et al., 1982). In humans, four AMPD isoforms have been

Abbreviations: AMPD, adenosine monophosphate deaminase; IMP, inosine

monophosphate acid; SNP, single nucleotide polymorphism; IMF, intramuscular fat; HF,
heart failure; CAD, coronary artery disease.
Corresponding author at: Department of Animal Genetics, Breeding and Reproduction,
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing
210095, PR China.
E-mail addresses: 2013205009@njau.edu.cn (J. Hu), 550619781@qq.com (P. Yu),
2011205009@njau.edu.cn (X. Ding), 2012105040@njau.edu.cn (M. Xu),
2012105038@njau.edu.cn (B. Guo), xuyinxue@njau.edu.cn (Y. Xu).

http://dx.doi.org/10.1016/j.gene.2015.08.008
0378-1119/ 2015 Elsevier B.V. All rights reserved.

identied: M (muscle); L (liver); and E1 and E2 (erythrocyte), which are

encoded by three genes (AMPD1, AMPD2 and AMPD3) of a multigene family (Sabina et al., 1990; Bausch-Jurken et al., 1992; Mahnke-Zizelman
et al., 1996). In Rattus, these isozymes are named the A (muscle), B
(liver and kidney), and C (heart) isoforms (Ogasawara et al., 1974),
respectively. The amino acid and nucleotide sequences of the three
AMPD genes are conserved and are expressed in a tissue-specic manner
(Mineo et al., 1990; Mahnke-Zizelman and Sabina, 1992; Gross et al.,
1994; Van den Bergh and Sabina, 1995). The AMPD1 gene is highly
expressed, predominantly in skeletal muscle. It is also the rate-limiting
step in the purine nucleotide cycle (Kuchiba-Manabe et al., 1991).
In humans, the most recent studies on the AMPD1 polymorphisms
have focused on the T34 allele. For example, previous studies detected
much lesser frequency of AMPD1 allele T and genotype TT in athletes engaged in the high-speed and strength sports (N = 305) in comparison
with humans practicing no sports (N = 499), then thought that the
AMPD1 gene polymorphism C34T can be considered to be a marker of
the liability for high-speed and strength related to muscular activity
(Fedotovskaya et al., 2013). The AMPD1 C allele may help athletes attain
elite status in sprint/power-oriented sports, whereas the T allele is
unfavorable for sprint/power-oriented athletic categories. Hence, the
AMPD1 C allele can be regarded as a marker associated with the sprint
and power aspects of physical performance (Ginevi et al., 2014). Several
reports have suggested the T34 allele to be associated with improved
outcome in patients with HF (heart failure) and improved cardiovascular survival in patients with CAD (coronary artery disease). However,
other reports did not conrm such associations (Kolek et al., 2005; de
Groote et al., 2006).

J. Hu et al. / Gene 574 (2015) 204209

Table 1
Chicken AMPD1 and -actin primer sequences and information.
Primer

Length (nt)

Primer sequence (53)

Temperature

P4-F/R

270

52

P6-F/R

429

P8-F/R

291

A4-F1/R1

431

A4-F2/R2

309

A6-F/R

429

A8-F/R

295

-actin-F/R

152

AMPD1-F-R

229

5-CCCTCTACTCTTGGCTATGT-3
5-ACTCCCTGAGACTCCTGTT-3
5-TGTGCCTGTCCTCTGTTGGG-3
5-TTTCAGGAGGAAGGGGCAAC-3
5-CGTTTCATCAAGAAGTCGT-3
5-AGAAGACAAGAGGCAAATC-3
5-CCCTCTACTCTTGGCTATGT-3
5-ACTCCCTGAGACTCCTGTT-3
5-TGAGGCTTTCCTTCCTGGCAGGTGG-3
5-TGTCTTCAGGCTGAGCATGCGCATT-3
5-TGTGCCTGTCCTCTGTTGGG-3
5-TTTCAGGAGGAAGGGGCAAC-3
5-CGTTTCATCAAGAAGTCGT-3
5-AGAAGACAAGAGGCAAATC-3
5-GAGAAATTGTGCGTGACATCA-3
5-CCTGAACCTCTCATTGCCA-3
5-TGTGGATGCTGACCGTGTA-3
5-ACCGTTGATGGTGTTTTCTGT-3

205

UV-754 photometer (SMOIF, Shanghai, China). Total RNA was stored

at 80 C while DNA was stored at 20 C until use. Three samples
of the same tissues were then mixed into an RNA-pool. Then, 0.32 g
of total RNA was used to synthesize rst-strand cDNA using the M-MLV
reverse transcriptase (Promega, Madison, USA) and oligo dT (18).

60
65
57
57
60
57
58
58

In pigs, the AMPD1 gene was mapped to SSC 4q1.6q2.3 (Stratil et al.,
2000). Previous studies indicate that the AMPD1 gene might be a
candidate gene for meat production traits and may provide useful
information for future studies in terms of its role in porcine skeletal
muscle (Wang et al., 2008). In cows, reports suggest that an 18-bp
deletion mutation in AMPD1 may inuence carcass traits. Also, two
SNPs (g.19416TNC and g.19421ANG) in the AMPD1 gene could be used
as markers for assisted selection in Qinchuan beef cattle-breeding
programs (He et al., 2010, 2011).
Few studies have provided complete genetic information on the
chicken AMPD1 gene. The objective of our study was to identify
AMPD1 gene polymorphisms and analyze the associations between
the SNPs and the IMP contents of Fast Partridge and Lingshan chickens.
This information may provide a possible candidate gene for markerassisted selection in chicken-breeding programs.

2.4. Screening for genetic variants in the coding region

Three primer pairs (P4-F/R, P6-F/R and P8-F/R, Table 1) were
designed using Premier 5.0 to clone the coding region based on the
GenBank reference sequence (GI: NC_006113.3). The primer pairs
were synthesized by Shanghai Sangon Biological Engineering Co., Ltd.
The DNA fragments amplied from 24 randomly selected samples
were directly sequenced in both directions using an ABI PRISM 3730
DNA analyzer (Applied Biosystems, USA) following standard procedures. The sequencing results were analyzed using the DNASTAR 5.0
software package (DNASTAR, Inc., USA) to detect genetic variations in
the AMPD1 gene.

2.5. Genotyping of the genetic variants

Sequencing revealed three genetic variations (g.4064G/A, g.5573A/G
and g.6805G/A) in exons IV, VI and VIII of the chicken AMPD1 gene. Four
primer pairs (A4-F1/R1, A4-F2/R2, A6-F/R and A8-F/R, Table 1) were
redesigned to be used for genotyping using Premier 5.0. g.4064G/A
was subsequently genotyped by bidirectional PCR amplication of specic alleles (Bi-PASA); and g.5573A/G and g.6805G/A were genotyped
by means of restriction fragment length polymorphism-PCR (RFLPPCR). The corresponding restriction endonucleases (AciI and MwoI)
were used to digest the PCR products. The digested fragments were
separated by 1.5% agarose gel electrophoresis with ethidium bromide
staining.

2. Materials and methods

2.6. Real-time PCR analysis of chicken AMPD1 mRNA levels in tissues

2.1. Source of animals

Real-time PCR was used to investigate the expression patterns of the

AMPD1 gene within multiple tissue types. The tissue types included:
heart, liver, spleen, lung, kidney, gizzard, glandular stomach, breast
muscle, leg muscle, subcutaneous fat, and abdominal fat. The -actin
gene was used as an endogenous reference for determination of
targeted mRNA proles, and was amplied using the -actin-F/R primer
pair (Table 1) (GenBank accession no. NM_205518). Primers for the
AMPD1 gene (AMPD1-F/R) were designed using Premier 5.0 (Table 1).
The reactions were performed in triplicate for each sample with 10 L
of 2 SYBRs Premix Ex Taq, 0.8 L of each primer, 0.4 L of 50 ROX
Reference Dye, 2 L of cDNA (100 ng/mL) and dH2O to a nal volume
of 20 L. PCR amplication was performed by conducting 42 cycles of
denaturation at 95 C for 10 s, annealing at 58 C for 30 s, and extension
at 72 C for 30 s, after the initial denaturation step (95 C for 3 min). PCR
product specicity was veried using a melting curve and by agarose gel
electrophoresis. No bands were seen in mock reactions from which
reverse transcriptase was omitted. The 2 Ct method was used to
identify differences in expression levels among the 11 tissues.

A total of 317 Fast Partridge (n = 157) and Lingshan chickens

(n = 160) was randomly selected from commercial populations
and used in the association analysis. The animals (60 10 days of age
at slaughter) were reared in Jiangsu Province, China. All experimental
procedures involving animals were carried out in compliance with
local legal regulations and all animal experiments were approved by
the Animal Care and Use Committee.
2.2. IMP quantication
The IMP content in chicken muscle (after ~24 h of storage at 4 C)
was measured by HPLC. A ZORBAX SB-C18 column (5 m,
4.6 250 mm) was used with a mobile phase A: 0.05 M triethylamine
phosphate and mobile phase B: methanol at a ow rate of 1 mL/min.
The column temperature was set at 25 C and UV detection was
performed at 254 nm.
2.3. Total RNA/genomic DNA isolation and rst-strand cDNA synthesis
Total RNA was isolated from chicken tissues using a TRIzol reagent
(Invitrogen, California, US) and treated with DNaseI (RNase-free)
(TaKaRa, Dalian, China) to remove genomic DNA. Genomic DNA was
extracted from pectoral muscle using the TIANamp Genomic DNA Kit
(Tiangen, Beijing, China) following the manufacturer's instructions.
The yield and quality of total RNA and DNA were determined spectrophotometrically using the 260/280 nm absorbance ratio with an

Table 2
Muscle IMP content in the two chicken breeds.
Breed

Number

Mean SE

Lingshan chicken
Fast Partridge chicken

160
157

1.8923 0.0480a
1.4484 0.0439b

Note: Values denoted by a,b in the same row are signicantly different at the 0.05 level
(P b 0.05).

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J. Hu et al. / Gene 574 (2015) 204209

Table 3
Muscle IMP content in different sexes of the same chicken breed.
Breed

Sex

Mean SE

Lingshan chicken

Cock (N = 80)
Hen (N = 80)
Cock (N = 78)
Hen (N = 79)

1.7714 0.0652b
2.0132 0.0641a
1.4079 0.0770
1.4875 0.0441

Fast Partridge chicken

Note: Values denoted by a,b in the same row are signicantly different at the 0.05 level
(P b 0.05).

2.7. Statistical analysis

Genotypic and allelic frequencies of each SNP, the Chi-square test for
HardyWeinberg equilibrium, the polymorphism information contents
(PICs), the heterozygosities (Hes), and the effective number of alleles
(Ne) were analyzed using the TFPGA software. Linkage disequilibrium
and haplotype analysis were performed using the SHEsis software.
Correlation analyses between the SNPs and IMP content were
performed by the least-squares method, using the SPSS software
(SPSS Statistics ver. 18.0), according to the following model:
Yi jk u Fi T j ei jk :

comparing sequences with that of the chicken AMPD1 gene (GI: NC_
006113.3). SNP g.4064A/G was found within exon IV, g.5573G/A within
exon VI, and g.6805A/G within exon VIII of the AMPD1 gene. All SNPs
were purine conversions that caused no alteration in the amino acid
sequence.
3.3. Genotyping the genetic variations by Bi-PASA and PCR-RFLP
The SNP g.4064A/G genotyped by Bi-PASA and the amplied AMPD1
gene g.4064A/G locus produced fragment sizes of: 431 bp and 309 bp
for genotype GG; 431 bp and 171 bp for genotype AA; and 431 bp,
309 bp and 171 bp for genotype GA (Fig. 1D). The SNPs g.5573G/A
and g.6805A/G were genotyped by PCR-RFLP in 117 chickens. Digestion
of the amplied AMPD1 gene g.5573G/A locus with AciI produced fragments of the following sizes: 429 bp for genotype AA; 109 bp, 320 bp
and 429 bp for genotype AG; and 109 bp and 320 bp for genotype GG
(Fig. 1E). Digestion of the amplied g.6805A/G locus with MwoI produced the following fragment sizes: 200 bp and 95 bp for genotype
GG; 95 bp, 200 bp and 295 bp for genotype GA; and 295 bp for genotype
AA (Fig. 1F). The obtained genotypes were in agreement with the DNA
sequencing results.
3.4. Genetic parameter analysis of three genetic variations

Yijk is the observed value; u is the overall mean; Fi is the effect of

farm; Tj is the effect of genotype; and eijk is the random error. Multiple
comparisons were performed using Fisher's least signicant difference
method. A value of P b 0.05 was taken to indicate statistical signicance.

The IMP content of Lingshan chicken was signicantly higher than

that of Fast Partridge chicken (P b 0.05) (Table 2). Also, in Lingshan
chicken the hen had a signicantly higher IMP content than did the
cock (P b 0.05), but no signicant difference between the hen and
cock was detected in Fast Partridge chicken (Table 3).

The A, G and A alleles are the predominant alleles of g.4064A/G,

g.5573G/A and g.6805A/G, respectively, in Lingshan chicken (Table 4).
The A allele is the predominant allele of the three loci in Fast Partridge
chicken (Table 4). Their high frequencies might be attributed to longterm breeding and selection for various purposes. Chi-squared tests
showed that g.5573A/G in Lingshan chicken and g.6805A/G in Fast Partridge chicken are in disagreement with HardyWeinberg equilibrium
(P b 0.05) (Table 4), whereas all others were in HardyWeinberg equilibrium (P N 0.05) for the analyzed population. According to the PIC classication (PIC value b 0.25, low polymorphism; 0.25 b PIC value b 0.5,
intermediate polymorphism; and PIC value N 0.5, high polymorphism),
all loci possessed intermediate polymorphism in the chicken AMPD1
gene in the analyzed population (Table 4).

3.2. Identication of genetic variants within the coding region of the AMPD1
gene

3.5. Associations between single variations and IMP contents in Fast

Partridge and Lingshan chickens

Three SNPs (g.4064A/G, g.5573G/A and g.6805A/G, Fig. 1A, B, C)

were detected in the samples by direct DNA sequencing and by

The effects of the three loci of the AMPD1 gene on IMP contents were
assessed for 317 chickens comprising Fast Partridge (n = 157) and

3. Results
3.1. IMP content

Fig. 1. AMPD1 sequencing data (A, B and C). Electrophoretic patterns of the chicken AMPD1 gene following resolution in 1.5% agarose gels (D, E and F). M: marker I; (D) genotypes:
GA = 431 bp, 309 bp and 171 bp; GG = 431 bp and 309 bp; AA = 431 bp and 171 bp; (E) genotypes: AA = 429 bp; GG = 320 bp and 109 bp; AG = 429 bp, 320 bp and 109 bp; and
(F) genotypes: GA = 295 bp, 200 bp and 95 bp; GG = 200 bp and 95 bp; and AA = 295 bp.

J. Hu et al. / Gene 574 (2015) 204209

207

Table 4
Genotype distribution, allelic frequencies and genetic diversity of the three SNPs in the AMPD1 gene.
Locus

g.4064G/A
g.5573A/G
g.6905G/A

Breed

Lingshan chicken
Fast Partridge chicken
Lingshan chicken
Fast Partridge chicken
Lingshan chicken
Fast Partridge chicken

Number

160
157
160
157
160
157

Genotype frequency

Gene frequency

GG

GA

AA

0.1695
0.1579
0.1695
0.1930
0.0339
0.0351

0.5254
0.6140
0.6949
0.5965
0.4576
0.6316

0.3051
0.2281
0.1356
0.2105
0.5085
0.3333

0.4322
0.4649
0.5169
0.4912
0.2627
0.3509

0.5678
0.5351
0.4831
0.5088
0.7373
0.6491

PIC

He

Ne

0.281
3.087
9.065
2.163
1.871
8.459

0.370
0.376
0.375
0.375
0.312
0.352

0.491
0.497
0.499
0.500
0.387
0.456

1.926
1.990
1.998
1.999
1.632
1.837

2 (HardyWeinberg equilibrium 2 value), He (gene heterozygosity), Ne (effective allele numbers), PIC (polymorphism information content).
In the same row are signicantly different at the 0.05 level (P b 0.05).

Lingshan chickens (n = 160) (Tables 5 and 6). In Lingshan chicken, the

polymorphisms at g.4064A/G and g.6805A/G were correlated with IMP
content (P b 0.05). Animals with the homozygous genotype AA at positions 4064 and 6805 had a signicantly greater IMP content than those
with the genotype GG (P b 0.05); and animals with the homozygous
genotype AA at position 6805 had signicantly higher IMP contents
than did those with the genotype GG in both cock and hen. The correlation between the g.5573A/G polymorphism and IMP content was nonsignicant for the 160 Lingshan chickens (P N 0.05). In Fast Partridge
chickens, the polymorphism at g.6805A/G was correlated with IMP content (P b 0.05). Animals with the homozygous genotype GG at position
6805 had a signicantly greater IMP content than did those with the
genotype AA (P b 0.05). Also, cocks with the homozygous genotype
GG at position 6805 had signicantly higher IMP contents than did
those with the genotype GG. The correlation between the g.4064A/G
and g.5573G/A polymorphisms and IMP contents were insignicant
for the 157 Fast Partridge chickens (P N 0.05). In conclusion, the polymorphism at g.6805A/G was correlated with IMP content (P b 0.05)
in both Fast Partridge and Lingshan chickens. Therefore, the SNP
6805A/G may be used as a molecular marker for the selection of
chickens with high IMP content. Furthermore, this study will facilitate
identication of a molecular marker of animals with better performance
for use in the chicken industry.
3.6. Tissue distribution of AMPD1
Real-time PCR was employed to examine AMPD1 expression in 11
tissues (heart, liver, spleen, lung, kidney, gizzard, glandular stomach,
breast muscle, leg muscle, subcutaneous fat and abdominal fat).
AMPD1 transcripts were detected at high levels in leg muscle, subcutaneous fat, and abdominal fat; at low levels in heart and kidney; and
were not detected in glandular stomach, spleen, and lung (Fig. 2).
4. Discussion
The IMP content is one of the main contributors to the avor and
aroma of muscle (Horio and Kawamura, 1990; Yamaguchi and
Ninomiya, 2000), while the AMPD1 can catalyze the hydrolytic deamination of AMP to IMP and an ammonium ion. Investigating the genetic
polymorphisms of the AMPD1 gene and their correlations with IMP

content in chicken is very useful in practice. For this aim, rstly, we

have analyzed the IMP content in different breeds and sex of chicken,
and then analyzed the associations between the SNPs and the IMP
content of chicken.
In the present study, we have investigated the Fast Partridge and
Lingshan chickens. Fast Partridge chicken is a cultivar crossed from
Partridge Shank chicken and Fast Big chicken, which is a breed of commercial broilers. While Lingshan chicken is the domesticated Lingshan
subspecies of jungle fowl, which is known to have a high protein
content and low fat and water-holding capacity as well as desirable
taste and avor. Our results revealed that the hen had a signicantly
higher IMP content than the cock (P b 0.05). Meanwhile, we found
that Lingshan chicken had a signicantly higher IMP content than Fast
Partridge chicken (P b 0.05), which is one of the reasons for the mellow
meat of native varieties (Chen et al., 2009; Liu et al., 2012). The data
suggested that different breeds could inuence the IMP content and
the native chicken may have a signicantly higher IMP content than
commercial broilers.
Chi-squared tests showed that the g.5573G/A in Lingshan chicken
and g.6805A/G in Fast Partridge chicken were in disagreement with
HardyWeinberg equilibrium (P b 0.05), whereas all other loci were in
HardyWeinberg equilibrium (P N 0.05) for the analyzed population.
This might be caused by the different effects of mutation and selection
pressure in different groups. And all loci possessed intermediate polymorphism in the chicken AMPD1 gene of the analyzed population,
which suggested that these loci have marked genetic variations, and
certain genetic diversity and potential ability of breeding.
All mutations (g.4064G/A, g.5573A/G and g.6805G/A) in chicken
AMPD1 were silent mutations and have been assumed to exert no discernible effect on gene function or phenotype as they do not alter coding
sequences and caused no alteration in amino acid sequence. However,
effects of the silent mutations on the gene function and phenotype
have been reported. A silent polymorphism in the MDR1 gene resulted
in a substrate specicity change (Kimchi-Sarfaty et al., 2007), and a
silent mutation in the goat POU1F1 gene was associated with milk
yield and birth weight (Lan et al., 2007). Other reports detailed the
effects of synonymous mutations in the gene at the protein level
(Shabalina et al., 2013). In this study, we identied the synonymous
mutations g.4064A/G, g.5573G/A and g.6805A/G, of which g.6805A/G
was signicantly associated with IMP content. Hence, it would be

Table 5
Least squares mean (LSM) + standard errors (SE) of IMP contents according to AMPD1 genotype in the two chicken breeds.
Breed

Lingshan chicken

Fast Partridge chicken

Locus

g.4064G/A
g.5573A/G
g.6905G/A
g.4064G/A
g.5573A/G
g.6905G/A

Genotype (LSM + SE)

AA

AG

GG

2.0717 0.7609a
1.8420 0.0823
2.0957 0.0491a
1.4469 0.0485
1.4260 0.0651
1.3277 0.0613b

1.9090 0.4440ab
1.9489 0.0602
2.0700 0.0400a
1.4964 0.0950
1.4595 0.0713
1.5337 0.0488ab

1.7350 0.1655b
1.8551 0.0532
1.7670 0.0698b
1.5291 0.0966
1.4909 0.0581
1.7090 0.0880a

Note: Values denoted by a,b in the same row are signicantly different at the 0.05 level (P b 0.05).

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J. Hu et al. / Gene 574 (2015) 204209

Table 6
Least squares mean (LSM) + standard errors (SE) of IMP content according to AMPD1 genotype between the sexes in the two chicken breeds.
Breed

Lingshan chicken

Locus

g.4064G/A
g.5573A/G
g.6905G/A

Fast Partridge chicken

g.4064G/A
g.5573A/G
g.6905G/A

Sex

Cock
Hen
Cock
Hen
Cock
Hen
Cock
Hen
Cock
Hen
Cock
Hen

Genotype (LSM + SE)

AA

AG

GG

2.0071 0.1107
2.1127 0.1048
1.8551 0.0532

1.9841 0.0633a
2.1723 0.0692a
1.4085 0.1898
1.5717 0.0760
1.4779 0.0980
1.4000 0.0874
1.1962 0.1189b
1.4044 0.0616

1.8744 0.0479
1.9460 0.0769
1.8426 0.1089
2.0102 0.0699
1.7250 0.1125b
2.0700 0.0400ab
1.4451 0.0852
1.4487 0.0520
1.4728 0.0829
1.4830 0.0692
1.5513 0.0766ab
1.5127 0.0599

1.7317 0.2531
1.7400 0.2130
1.7901 0.1230
1.8940 0.1211

1.8007 0.0902b
1.6004 0.1278
1.4398 0.3101
1.3538 0.3057
1.5242 0.0669
1.6209 0.0332a
1.7970 0.1406

Note: Values denoted by a,b in the same row are signicantly different at the 0.05 level (P b 0.05).

interesting to identify the mechanism underlying the association between these silent mutations and the IMP contents of Lingshan, Fast
Partridge, and native chickens.
In our study, in Lingshan chicken the polymorphisms at g.4064A/G
and g.6805A/G were correlated with IMP content (P b 0.05). Animals
with the homozygous genotype AA at positions 4064 and 6805 had
signicantly higher IMP contents than those with the GG genotype
(P b 0.05). Also, the homozygous genotype AA at position 6805 had a
signicantly higher IMP content than those with the genotype GG for
both cock and hen. The correlation between the g.5573G/A polymorphism and IMP content was non-signicant (P N 0.05). In Fast Partridge,
the polymorphism at g.6805A/G was correlated with IMP content
(P b 0.05). Animals with the homozygous genotype GG at position
6805 had signicantly higher IMP contents than those with the
genotype AA (P b 0.05). Also, hens with the homozygous genotype GG
at position 6805 had signicantly higher IMP contents than did cocks
with the genotype GG. The correlation between the g.4064A/G
and g.5573A/G polymorphisms and IMP content was non-signicant
(P N 0.05). Statistical analysis revealed that Lingshan chicken with the
homozygous genotype AA at position 6805 had a signicantly greater
IMP content, however, Fast Partridge chicken with the genotype GG at
position 6805 had a signicantly greater IMP content. It is interesting
to note that the genotypes are different in Lingshan and Fast Partridge
chickens with a signicantly greater IMP content, which might be attributed to Lingshan chicken being the native chicken and Fast Partridge
chicken as a breed of commercial broilers (a cultivar crossed from
Partridge Shank chicken and Fast big chicken). However, the mechanism underlying this phenomenon is still not clear, therefore, further
research will be explored to investigate the differences in the

Fig. 2. Expression proles of chicken AMPD1 mRNA in 11 tissue types, determined in triplicate. The mean value in breast muscle was set as 1.

relationship of genotypes and IMP content between other native

chickens and commercial broilers.
Conict of interest
There is no conict of interest.
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