Position of the Phi and Po2 loci in the Hal linkage group in pigs

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Position of the Phi and Po2 loci in the Hal linkage

group in pigs

P. Vögeli

To cite this version:

Original

article

Position of the Phi and Po2 loci in the Hal

_linkage

_group

in

_pigs

P.

_Vögeli

Insfifute of Animal Sciences,

Breeding

Section, Federal Institute of

_Technology

_(ETH), CH-8092

Zurich, Switzerland

(received

17

_May

_1988,

_accepted

8 October

1988)

.

Summary —

Families of Swiss Landrace

₍₁₆₅

litters with 1348

_offspring)

were tested for halothane

sensitivity, A-0(S),

H, Phi, PGD and Po2

_phenotypes.

Informative

matings

for the determination of the _{gene sequence of these} linked loci were selected. Recombinations were observed between

Phi-Hal, Phi-H and H-Po2. On the basis of these results the most

_likely

order. of loci is Hal-Phi-H. Confirmation for a locus _{for genes for Po2 separate} from the locus for H is

presented.

The location of Po2 is between H and

Pgd.

A _{gene order}

_{S-Hal-Phi-H-Po2-Pgd}

is

_proposed.

pig - Iinkage

group -

S-Hal-Phi-H-Po2-Pgd -

halothane

sensitivity -

gene order

Résumé — La

_position

des loci Phl et Po2 dans le _groupe de liaison S,

Met

Phl,

H,

PoZ

Pgd

des porcs. Des familles de Porc amélioré suisse

(165 portées

avec 1348

descendants)

ont été tes-tées pour la sensibilité à 1 halothane, ainsi que pour les

phénotypes

A-O

(S),

H, PHI, PGD, et Po2. Des

_{accouplements}

_{informatifs pour déterminer l’ordre} de ces loci liés ont été sélectionnés. Des recombinaisons ont été trouvées entre Phi-Hal, Phi-H et H-Po2. Ces résultats

_{ont permis}

de

préci-ser la

position

du locus Phi dans _{le groupe de liaison. L’ordre le}

plus probable

des loci est Hal-Phi-H. Ces données confirment que les

_gènes

Po2 et H se situent à deux loci distincts. Po2 se situe

entre H

_{et Pgd.}

En conclusion

_{est proposé Ibrdre génique S-Hal-Phi-H-Po2-Pgd.}

porcs - groupe de Ilaison -

S-MeM

W

-H-Po2-Pgd -

sensibilité à l’halothane - ordre de loci

Introduction

The

_linkage

between the H blood group locus and the loci for the variants of

6-phospho-gluconate dehydrogenase

(Pgtn

and

_{phosphohexose}

isomerase

_(Phi)

was first described

by

Andresen

₍₁₉₇₁

_{). Rasmusen}

& Christian

₍₁₉₇₆₎

reported

an association between

H

_genotypes

and

_{susceptibility}

to halothane-induced stress.

_Jorgensen

et al.

₍₁₉₇₆₎

pos-tulated that the association between H and

_porcine

stress and the

_linkage

of Phi and H

was the causal link for the association between Phi

_genotypes

and stress

_{susceptibility.}

The inheritance of halothane-induced stress has been shown to be controlled

_by

a

Bampton, 1977).

N

_symbolizes

Hal N and n, Hal n.

_Pigs

that are N/N or Nln should there-fore be HAL- non reactors and nln

_signifies

a HAL+reactor.

Linkage

studies between Hal and Phi have not made it

possible

to

_place

the Hal locus

accurately

within the

_linkage

_group.

_Using

a method for calculation of relative

_linkage

disequilibrium

coefficients,

Andresen

(1979) proposed

that Halwas located between Phi and H. The gene order

Phi-Hal-H-Pgd

was also

supported by

Rasmusen et al.

(1980).

Their

_{data, however,}

did not

_permit

them to

_distinguish

between the order

_{Phi-Hal-H-Pgd}

as

_opposed

to

Hal-Phi-H-Pgd.

Gu6rin et al.

(1983)

described two recombinants which

supported

the order as

_Hal-Phi-Pgd.

The recombinants were both

_{HAL- offspring}

of

matings

between Hal Nln _{and Hal} Wn _{animals. The failure of these animals to react to}

halo-thane

could, however,

have resulted from the

_incomplete

penetrance

of the Haln gene. The S locus controls the

_expression

of the A and 0

_antigens

of the A-O blood _group

system

in

_{pigs by}

an

_epistatic

interaction

_(Rasmusen,

1964 and

_Hojny

&

Hdla,

1965).

Two alleles are

_known,

S

_being

dominant over s.

The

_relationship

between A-O _{blood group}

_phenotypes

determined

_by

_genes at the S

locus and HAL+ animals

(Rasmusen

&

_{Christian, 1976)}

was in

_agreement

with the

asso-ciations found between A-O and H blood group

systems

(Rasmusen, 1972).

Hojny

(1974)

suggested

that this association

resulted,

indirectly,

from the

genetic linkage

between the H

_system

and the S locus. Rasmusen

_{(1981) proposed}

the order

_{Phi-Hal-S-H-Pgd on}

the basis of recombinants between S and H as well as between S and Phi-Hal.

Later,

two

.other

reports

provided

evidence that the S locus is not within the

_{Phi-Pgd region,}

but

adjacent

to Phi

_(Hojny

et

al., 1985;

Van Zeveren et

al.,

1985).

-Recently

it has been found that the serum

_{postalbumin-2 (Po2 locus)}

also

_belongs

to

the S-

_{(Phi-Hal-H)-Pgd}

_linkage

_{group and is}

_probably

located between the H and

_Pgd

loci

_(Juneja

et al., 1983;

Gahne &

_Juneja,

1985;

Cepica

et al.,

1986).

The aim of this paper is to _{reconsider the gene order in the}

_linkage

_group,

_especially

of the Phi and Po2 loci and to establish the

haplotypes (including

Hal

_genotypes)

in a

population

of Swiss Landrace

_pigs.

Estimation of recombination

_frequencies

is

_given

elsewhere

_(V6geli

et al.,

1988).

Materials and Methods

Description

of the data

Data for this

_study

came from Swiss Landrace

pigs kept

at the

_experimental

station of the Institute of Animal Sciences

_during

the

period

1983-1988. The total number of

_offspring

was 1348. The animals came from 165 litters

_{produced by}

29 boars and 64 sows over 3

successive

_generations.

Halothane test

At an _{age of 8} to 12 weeks the animals were tested for halothane

_{sensitivity by}

the

method of Eikelenboom & Minkema

_(1974).

The anesthetic was a _{mixture of oxygen and}

4% halothane

_{(1.5 liters/min). Negatively reacting}

animals were

_exposed

for 5 min. In

HAL

+

_pigs

the anesthesia was withdrawn as soon as the

symptoms

of

_hyperthermia

(muscular rigidigy,

increased heart rate and elevated

body temperature)

became

Serological

tests

The A and 0

_reagents

were

_prepared

from normal serum of 2

_goats

and were used in the

hemolytic

test. The alloimmune anti-Aw was

_applied

in the dextran

_{agglutination}

test.

The blood _group factors Ha and Hc were tested

_using

two

_reagents

each. One of each

exhibited

_dosage

_{effects, i.e.,}

_{they hemolysed}

red blood cells of

_homozygous

_(H

_a

/ h

O

,

H C_{/ H}

C

)

pigs

sooner than those derived from

_heterozygous

_{(H a / H -, H C / H -)}

_pigs.

The

_validity

of the reaction

_pattern

of these

_reagents

was verified in International

_Pig

Comparison

Tests

₍₁₉₈₄

and

1987,

_the

latter

_{being organized by}

our

_laboratory).

Electrophoresis

-The Phi and

_{Pgd phenotypes}

were determined

by

horizontal one-dimensional agarose

or

_starch-gel

_{electrophoresis}

of

hemolysates

of

_erythrocytes

_(Saison

&

Giblett, 1969;

Gahne &

_{Juneja, 1985).}

The Po2 variants were detected

_by

two-dimensional

electropho-resis

_by

the method of

_Juneja

era/.

_(1983).

Parentage

control

Tests for other blood marker

_systems

_(B,

G, ADA, PGM, P11, P01A,

P12 )

were conduc-ted on all animals for the exclusion of incorrect

_pedigrees.

Haplotyping

The method used in the

_present

_study

to determine the

_haplotypes

was based on

dedu-cing linkage

phases involving

Hal and marker loci of both the

parents

and their

_offspring.

A detailed

_description

of the

procedure

is

_given

_{by V6geli}

et al.

_(1988).

Several instances of

_crossing

over were _{observed in progeny from}

_{multiheterozygous}

_parents

mated to

multihomozygous

parents.

These were used to determine the order of the loci.

Results

Table

I provides

a _{summary of recombinations}

_involving

the Hal and Phi loci recovered in

progeny from

_matings

in which one

_parent

was at least

_{triply heterozygous}

and the other

doubly

or

_{multiply homozygous.}

All the recombinations are informative with

_respect

to the location of the Phi locus. The structure of

_{parental haplotypes}

was inferred from various informative

_matings.

Assuming

that the _gene order is Phi-Hal-H as

_{suggested by}

Rasmusen

₍₁₉₈₁₎

and not

Hal-Phi-H,

the first 5 recombinants of the first 3

_{matings given}

in Table I would have

required

the occurrence of a double crossover,

i.e.,

a crossover between Phi and Hal as

well as a crossover between Hal and H which is

statistically extremely unlikely.

Mating

of boar 8888 with female 8849

_produced

a recombinant

_(offspring

9925)

resul-ting

in an

_unexpected

halothane

_negative

reaction of this

_offspring.

This recombinant

could be

explained

as

_being

a result of double crossover.

However,

incomplete

penetran-ce of HaM / HaM seems more

_{likely. Unfortunately,}

the recombinant

_offspring

9925 was

not saved for

_breeding

to determine his actual

_genotype.

In the

_offspring

of animals with Hafn Hain

_genotype

mated to Ha! /

HaM,

about 10% are classified as HAL-

_(Gahne

&

Table II shows most informative recombinants between

S, Hal,

Phi and H on one side and Po2 and

_Pgd

on the other. All five recombinants are informative in

_determining

the

position

of the Po2 locus. From these data the _gene order is

_{H-Po2-Pgd as}

_{proposed by}

Juneja

et al.

_(1983).

_{If the gene order} were

_Po2-H-Pgd,

all 5 recombinants could

_only

have resulted from double crossovers

_(Phi

Î-P

0

2 Î-H-Pgd),

which is

_{highly improbable.}

Table III shows the

_parents

and

_offspring

of 2 litters which include recombinants

invol-ving

a crossover between loci for Phi and H

_types.

These marker loci are also consistent with a _{gene order of Phi-H-Po2} as

_opposed

to H-Phi-Po2.

Discussion

The

_{expected Hal genotype}

of

_{offspring receiving}

(a)

recombinant

_haplotype

(s)

can be determined if the sequence between Hal and marker loci has been established. The

most

likely

order of the marker loci

including

Hal was indicated as

_S-(Phi-Hal)

-(H-Po2)-Pgd by

Hojny

et al.

₍₁₉₈₅₎

and van Zeveren et al.

_(1985).

As these authors did not detect

sequence for the Phi and Hal loci

proposed by

Gu6rin et al.

_(1983).

However,

they

confir-med that the two loci are _{located very close} to each other.

The most

_important

_{contribution of this paper is the evidence that the Phi locus is}

located,

most

_probably,

between Hal and H as

_{proposed by}

Guérin et al.

₍₁₉₈₃₎

and van

Zeveren et al.

₍₁₉₈₈₎

and not between S and Hal as

_{previously reported}

_by

Andresen

(1981)

and Rasmusen

_(1981).

This location is more

_firmly

established

_by

_complex

S-Hal-Phi-H-Po2-Pgd haplotypes

of the

_majority

of

_parents

and

_{offspring, including}

recom-binants.

Probably

because of

incomplete

penetrance

of the Hal gene one animal with

offspring

275 and

_{695) being}

informative with

_respect

to the location of the Phi locus

were classified as HAL+. These two reactors

_certainly

are Hal !l Hal !

_homozygotes

because the

_probability

of a Hah l HaIN or HalN / Hain

_{pig being}

_falsely

tested as HAL+ is very low

(V6geli

et

_al.,

_1988).

The data in Tables II and III are consistent with a _{gene order of}

_{Phi-H-Po2-Pgd.}

The

data assembled in Tables

I,

II and III and

those

contained in earlier

_publications

indicate

a _{gene order}

_{S-Hal-Phi-H-Po2-Pgd.}

The

_knowledge

of the halothane locus and its linka-ge

relationships

is

_{already being}

used in

_practical

animal

_breeding

to reduce the

frequen-cy of the Hal ! gene

(Gahne

&

_Juneja,

1985;

V6geli

et

al.,

1988). Looking

to the

future,

molecular

_analysis

of the halothane

_linkage

_{group may}

_provide

a means for

_identifying

more reliable markers for the stress _genes as well as the

_identity

_{of the halothane gene} itself. One

step

in this

_development

is the

_assignment

of the Hal

_linkage

_group to

chro-mosome 6

_by

in situ

_{hybridization (Davies}

ef al.,

1988).

Acknowledgements

This work was

_{supported by}

grants of the ETH-Zurich, the Commission for

_Support

of Scientific

Research, Berne, the Swiss Performance

Testing

Station,

Sempach,

and other

public

and

private

organizations

in Switzerland.

_Appreciation

is

_expressed

to _{my coworkers Christine} Karonis, Barbara Kuhn and R. Kuhne for excellent technical assistance and Drs. N.S. Fechheimer, Valerie Madison,

References

Andresen E.

₍₁₉₇₁₎

_{Linear sequence} of the autosomal loci PHI, H and 6-PGD in

_pigs.

Anim. Blood

Groups

Biochem. Genet. 2, 119-120

Andresen E.

₍₁₉₇₉₎

Evidence

_indicating

the sequence Phi, Hal, H of the three

_dosely

linked loci in

pigs.

Nord Veterinaermeal. 31, 443-444

Andresen E.

₍₁₉₈₁₎

Evidence for a five-locus

linkage

group

involving

direct and associative interac-tions with the A-O blood group locus in

pigs.

In :

_(E.

Brummerstedt

_{ed.), Papers}

dedicated to Prof. Dr. J.

_Moustgaard,

208-212. The

_Royal

Danish

_{Agricultural Society, Copenhagen.}

!epica

S.,

Hradecki

J.,

Hojny

J.,

Kuryl

J. &

_Grzybowski

G.

₍₁₉₈₆₎

Localization of the Po2 locus in the S, Phi, Hal, H, Po2,

Pgd linkage

group in

_pigs.

Anim. Genet. 17, 283-286

Davies W., Harbitz L, Fries R.,

Stranzinger

G. &

_Hauge

J.G. (1988) Porcine

_{malignant hyperthermia}

carrier detection and chromosomal

_{assignment using}

a linked

_probe.

Anim. Genet. 19, 203-212 2 Eikelenboom G. & Minkema D.

₍₁₉₇₄₎

Prediction of

_pale,

soft, exudative muscle with a non-lethal

test for the halothane-induced

porcine malignant hyperthermia syndrome. Tijdschr. Diergeneesk.

99,

421-426

Gahne B. &

_Juneja

R.K.

₍₁₉₈₅₎

Prediction of the halothane

(Hal )

genotypes of

_{pigs by deducing}

Hal, Phi, Po2,

Pgd haplotypes

of parents and

offspring:

results from a

_{large-scale practice}

in Swedish breeds. Anim. Blood

_Groups

Biochem. Genet. 16, 265-283

Gu6rin G., Ollivier L. & Sellier P. (1983) Etude de groupe de liaison Hal, Phi et

Pgd

chez le Porc:

disposition

relative des trois locus et estimation des taux de recombinaison. Genet. Sel. Evol. 15,

55-64

Hojny

J.

(1974)

H _{blood group genotypes} and

_expression

of A and 0

_antigens

in

_pigs.

Anim. Blood

_Groups

Biochem. Genet. 5, 3-10 0

Hojny

J. & Hila K.

₍₁₉₆₅₎

A contribution to the

_study

of the blood _{group system} A in

_pigs.

In: Blood groups of animals

(J.

Matousek ed.).

Publishing

House of the Czechoslovak

Academy

of Sciences,

Prague. pp.155-161

Hojny,

Cepica

S. &

_Hradecky

J.

₍₁₉₈₅₎

Gene order and recombination rates in the

_linkage

_group

S-Phi-Hal-H-(Po2)-Pgd

in

_pigs.

Anim. Blood

_Groups

Biochem. Genet. _16, 307-318 8

Juneja

R.K., Gahne B.,

Edfors-Lilja

I. & Andresen E.

₍₁₉₈₃₎

Genetic variation at a

_pig

serum

_protein

locus, Po-2 and its

_assignment

to the Phi, Hal, S, H,

Pgd linkage

group. Anim. Blood

Groups

Bio-chem. Genet. 14, 27-36

Jorgensen

P.F.,

Hyldgaard-Jensen

J.,

Moustgaard

J. & Eikelenboom G.

(1976) Phosphohexose

iso-merase

_(PHI)

and

_porcine

halothane _sensitivity. Acta Vet. Scand. 17, 370-372

Ollivier L., Sellier P. & Monin G.

₍₁₉₇₅₎

D6terminisme

_g6n6fique

du

syndrome d’hyperthermie

maligne

chez le porc de Pi6train. Ann. Genet. Sél. Anim. 7,159-166

Rasmusen B.A.

₍₁₉₆₄₎

Gene interaction and the A-O blood _{group system} in

_pigs.

Genetics 50,

191-198

Rasmusen B.A.

(1972)

Gene interaction and the A-O and H _{blood group systems} in

_pigs.

Anim. Blood

Groups

Biochem. Genet. 3, 169-172

Rasmusen B.A. ₍₁₉₈₁₎

_Linkage

of _genes for PHI, halothane

_sensitivity,

A-O inhibition, H red blood cell

antigens

and 6-PGD variants in

_pigs.

Anim. Blood

Groups

Biochem. Genet. 12, 207-209

Rasmusen B.A. & Christian L.L.

₍₁₉₇₆₎

H blood _types in

_pigs

as

_predictors

of stress

_{susceptibility.}

Science 119, 947-948

Rasmusen B.A., Beece C.K. & Christian L.L.

₍₁₉₈₀₎

Halothane

_sensitivity

and

_linkage

of _{genes for} H red blood cell

_{antigens, phosphohexose}

isomerase

_(PHI)

and

_{6-phosphogluconate}

dehydrogena-se

_(6-PGD)

variants in

_pigs.

Anim. Blood

Groups

Biochem. Genet. 11, 93-107

Saison R. & Giblett E.R.

_{(1969) 6-phosphogluconic dehydrogenase polymorphism}

in the

_pig.

Vox

_Sanguinis

16, 514-516 6

Van Zeveren A., Van de

_Weghe

A.,

Bouquet

Y &

_Varewyck

H.

₍₁₉₈₅₎

The

_position

of the

_epistatic

S locus in the halothane

linkage

group in

_pigs.

Mim. Blood

_Groups

Biochem. Genet.16, 297-305

Van Zeveren A., Van de

_Weghe

A.,

Bouquet

Y. & Varewyck H. (1988) The

porcine

stress

_linkage

group. 11. The

position

of the Halothane locus and the _accuracy of the Halothane test

diagnosis

in

Belgian

Landrace

pigs.

J. Anim. Breed. Genet. 105, 187-194

’

Vögeli

P.,

Gerwig

C. & Schneebeli H.

₍₁₉₈₃₎

The A-O and H _{blood group systems,} some _enzyme

systems and halothane

_sensitivity

of two

_divergent

lines of Landrace

pigs using

index selection pro-cedures. Livest Prod. Sci. 10, 159-169