CHARACTERISATION OF SOME FATNESS CANDIDATE GENES IN BASQUE BLACK PIED AND LARGE WHITE PIGS (CARACTERIZACIÓN DE ALGUNOS GENES CANDIDATOS DE LA ADIPOSIDAD EN CERDOS PÍO NEGRO DEL PAÍS VASCO Y LARGE WHITE)

Alfonso L.

Description

Abstract
Characterisation of some fatness candidate genes in Basque Black Pied and Large White pigs.

Sujets

• Heart-type fatty acid binding protein
• LEPR
• LEP

Informations

• Publié par: erevistas
• Publié le: 01 janvier 2004
• Nombre de lectures: 11
• Langue: English

Extrait

NOTA BREVE
CHARACTERISATION OF SOME FATNESS CANDIDATE GENES
IN BASQUE BLACK PIED AND LARGE WHITE PIGS
CARACTERIZACIÓN DE ALGUNOS GENES CANDIDATOS DE LA ADIPOSIDAD EN
CERDOS PÍO NEGRO DEL PAÍS VASCO Y LARGE WHITE
Alfonso, L. and A. Arana
U.P.N.A. Departamento de Producción Agraria. 31006 Pamplona. Spain. E-mail: [e-mail]
PALABRAS CLAVE ADICIONALESADDITIONAL KEYWORDS
Adipose development. H-FABP, A-FABP, LEPR, Desarrollo adiposo. H-FABP, A-FABP, LEPR,
LEP. LEP.
INTRODUCTION
Fatness is a highly heritable ment. Several candidate genes, mapped
quantitative trait strongly influenced functional genes related to the expre-
by the environment, so only mutations ssion of a trait, have been suggested to
with very large effects are likely to be explain pig fatness. Four of them are
detected (Keightley, 1995). Breeds characterised in this work: the Heart
with a high adipose development offer Fatty Acid-Binding Protein (H-FABP),
an opportunity for this detection. An the Adipocyte Fatty Acid-Binding
example is the Basque Black Pied, one Protein (A-FABP), the Porcine Leptin
of those pig breeds that have lost its Receptor (LEPR) and the Porcine
productive role during the last century Leptin (LEP). Associations between
(Iriarte and Alfonso, 2000). The them and intramuscular fat content and
Basque breed exhibited an early and backfat thickness have been found in
higher adipose development and a different studies (Estany et al., 2002;
higher activity of enzymes responsible Gerbens et al., 1998; Gerbens et al.,
for lipid synthesis than selected pig 1999; Kennes et al., 2001).
populations (Alfonso et al., 2005).
Laval et al. (2000), in a study of pig
genetic diversity, indicated that the MATERIALS AND METHODS
Basque breed appeared to be the most
unique in the set of eleven pig breeds Samples from forty-two animals
originating from six European coun- were available from a previous
tries they analysed. So, the Basque experiment carried out to characterise
Black Pied can be considered as an the adipose development of Basque
interesting pig population to analyse pigs (n=20) by comparison with a
candidate genes of fat tissue develop- commercial selected line of Large
Arch. Zootec. 53: 411-414. 2004.ALFONSO AND ARANA
White origin (n=22) (Alfonso et al., percent HindIII) and the bands
2005). All these animals were geno- visualised with ethidium bromide.
typed for A-FABP microsatellite and Polymerase chain reaction conditions
the following RFLP polymorphisms: for amplification of the A-FABP
HinfI, HaeIII and MspI for the H-FABP microsatellite were as described by
gene, HinfI, HpaII and RsaI for the Gerbens et al. (1998). Resulting
LEPR gene and HindIII for the LEP fragments were detected on an ABI
gene. PRISM 310 Genetic Analyzer (ABI,
HinfI and HaeIII H-FABP polymor- Perkin Elmer, USA) and analysed using
phism were typed using the primers Genescan software (Applied Biosys-
described by Gerbens et al. (1997). tems, Perkin Elmer, USA).
Primers for MspI were Fw: 5’-ATT Allele and genotype frequencies
GCT TCG GTG TGT TTG AG-3’ and were determined by gene counting. A
Rev: 5’-GGC CAT CCC ATA GAA Chi-square test was carried out to com-
CT-3’. The PCR mixture contained 2 pare genotype frequencies among breeds.
mmol MgCl2, 0.2 mmol of each dNTP,
6 pmol of each primer, 100 ng genomic
RESULTS AND DISCUSSIONDNA, 0.7 U of Taq DNA polymerase
in a 25 µl final volume. HinfI LEPR
Allele frequencies estimated for thepolymorphism was typed using the
polymorphism analysed are shown inprimers described by Vincent et al.
table I. Biallelic polymorphisms were(1997), and HpaII and RsaI polymor-
found with HaeIII and MspI for H-phism with those described by Stratil
FABP gene, with HpaII and HinfI foret al. (1998). The PCR mixture
LEPR gene and with HindIII for LEPcontained 1.5 mmol MgCl2 (2 for
gene. For HinfI H-FABP and RsaIHinfI), 0.2 mmol of each dNTP, 15
LEPR polymorphisms the same allelepmol of each primer, 100 ng genomic
was found fixed in both breeds.DNA, 0.7 U of Taq DNA polymerase
Differences in genotype frequenciesin a 25 µl final volume. HindIII LEP
among breeds were significant for MspIpolymorphism was typed using the
H-FABP, HinfI LEPR and HindIII LEPprimers described by Kennes et al.
polymorphisms (table I). The C MspI(2001). The PCR mixture contained 1
H-FABP allele frequency was lower inmmol MgCl2, 0.2 mmol of each dNTP,
5 pmol of each primer, 75 ng genomic Basque than in Large White line. The
DNA, 0.2 U of Taq DNA polymerase a HinfI LEPR allele was fixed in
in a 25 µl final volume. Basque but segregating, although at a
PCR products were digested with 3 high frequency, in Large White. The A
U of the respective enzyme (37º C/3 HindIII LEP allele was segregating in
h.) and after PCR amplification Basque and absent in Large White.
fragment lengths were determined Results for the A-FABP micro-
upon electrophoresis on agarose gels satellite polymorphism analysed also
(H-FABP: 2 percent HinfI and HaeIII, showed significant differences among
3 percent MspI; LEPR: 1 percent HpaII, populations. Alleles A3 and A5 wereHinfI and RsaI; LEP: 3 found in Large White but absent in
Archivos de zootecnia vol. 53, núm. 204, p. 412.FATNESS CANDIDATE GENES
Basque, and frequencies in A and A (0.96), Torbiscal Iberian (0.8) and
2 4
alleles were markedly different, Black Hairless Iberian (1) populations
especially for A allele (0.88 in Basque (C. Ovilo, personal communication).
4
and 0.07 in Large White pigs). Second, whereas the a HinfI LEPR
To our knowledge, this study reports allele has been found fixed in Basque
the first results regarding the charac- pigs it was found to be segregating in
terisation of the Basque pig breed for Landrance, Duroc, Hampshire and
H-FABP, A-FABP, LEPR and LEP Meishan populations (Vincent et al.,
genes. The results indicate that Basque 1997) like in the Large White analysed
breed shows differences with the Large in the present study. Third, Kennes et
White line analysed for all these genes. al. (2001) found that A HindIII LEP
Differences can be also noticed in allele was segregating in Duroc and
comparison with the characteristics Landrace like observed in Basque
reported on other populations. First, breed, but at a lower frequency. Finally,
the C MspI H-FABP allele frequency it should be noticed that Ovilo et al.
was lower in Basque breed than values (2001) also found the A and A A-
2 4
estimated in Pietrain (1), Landrace FABP alleles in a population of Iberian
Table I. Allele frequencies at the H-FABP, LEPR, LEP and A-FABP gene polymorphisms and
Chi-square test significance of genotype frequencies comparison among breeds. (Frecuencias
alélicas para los polimorfismos de los genes H-FABP, LEPR, LEP y A-FABP y significación de la prueba
Chi-cuadrado de la comparación de las frecuencias genotípicas entre razas).
Breed Significance of
Basque Large White differences in
a bGene Polymorphism Allele Allele frequency Allele frequency genotype frequencies
H-FABP HinfI H 11 ns
HaeIII D 0.58 0.46 ns
MspI C 0.28 0.98 ***
LEPR HpaII b 0.65 0.68 ns
RsaI d 11 ns
HinfI a 1 0.70 ***
LEP HindIII A 0.73 0 ***
A-FABP Microsatellite A 0 0 ***
1
A 0.12 0.61
2
A 0 0.07
3
A 0.88 0.07
4
A 0 0.25
5
aH allele: 350+180+110+50+37 bp; D allele: 684+117+16 bp; C allele: 322+90 bp; b allele: 1450+550 bp;
d allele: 750+349+334+300+250 bp; a allele: 2100+700+395+240+140+110 bp; A allele: 658 bp; A allele
1
248 bp; A allele 250 bp; A allele 252 bp; A allele 254 bp; A allele 268 bp.
2 3 4 5
b 2χ statistic significance. ns: not significant; ***p<0.001.
Archivos de zootecnia vol. 53, núm. 204, p. 413.ALFONSO AND ARANA
pig, but with intermediate frequencies have been reported could allow the
between the values found in the Basque design of future experiments interested
and Large White breeds in the present in dealing with fatness genetic back-
study. ground, especially involving H-FABP,
The differences among breeds that A-FABP and LEP genes.
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