Gene mapping in the river buffalo (Bubalus bubalis L)

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Submitted on 1 Jan 1991

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Gene mapping in the river buffalo (Bubalus bubalis L)

Ha de Hondt, Aa Bosma, M den Bieman, Na de Haan, Lfm van Zutphen

To cite this version:

Gene

_mapping

in the river buffalo

(Bubalus

bubalis

_L)

HA de

Hondt

AA Bosma

M

den Bieman

NA de Haan

LFM

van

Zutphen

3

!

National Research

Centre,

Unit

_of

Cell

_{Biology, Dokki, Egypt;}

2

Utreciat

University, Faculty of Veterinary Medicine,

Department

of

Cell

Biology

and

_Histology,

and 3

Department of Laboratory

Animal

_Science,

The Netherlands

(Proceedings

of the 9th

_European

_Colloquium

on

_Cytogenetics

of Domestic

_Animals;

Toulouse-Auzeville,

10-13

_July

₁₉₉₀₎

river buffalo

_/

_{gene mapping}

_/

somatic cell hybrids

/

enzymes

/

enzyme gene loci

Somatic cell

_{hybridization}

was used as a tool to examine

_synteny

_{of genes} in the buffalo. Parental cells were blood

_lymphocytes

from two

_Egyptian

river

_buffaloes,

and cells of the HPRT-

_{(hypoxanthine}

_{phosphoribosyltransferase-negative)}

Chinese

hamster cell line

_wg3h

C

1

2

(Echard

et

al,

1984).

Hybrid

cells were

produced by

po-lyethylene

glycol

(PEG)-mediated

fusion,

followed

_by

hypoxanthine-aminopterine-thymidine

(HAT)

selection.

_{Twenty-one independent hybrid}

cell lines were

esta-blished. For enzyme

analysis,

hybrid

cells and hamster cells were

lysed

in

lysis

buffer

_(Meera

_Khan,

_1971).

Buffalo cardiac

_muscle,

skeletal muscle and

erythro-cytes were

homogenized

in the same

buffer,

and were used as

samples

of reference

for buffalo enzyme

activity.

Starch

gel electrophoresis

(Harris

and

Hopkinson,

1978)

was carried out to

_analyze

the

_following

_enzymes:

_{glyceraldehyde-3-phosphate}

dehy-drogenase

(GAPD;

EC

_1.2.1.12);

lactate

_{dehydrogenase}

_(LDH;

EC

_1.1.1.27);

malic

enzyme

(ME;

EC

1.1.1.40);

phosphoglucomutase

(PGM;

EC

5.4.2.2);

superoxide

dismutase

_(SOD;

EC

_1.15.1.1.);

_{triose-phosphate}

isomerase

_(TPI;

EC

_5.3.1.1).

Two different buffer

_systems

were used for the

_separation

of hamster _enzymes from

buf-falo enzymes: a

Tris/borate/EDTA (ethylenediaminetetraacetic acid)

_system,

pH

8.6,

(Shows

et

_al,

₁₉₆₄₎

for

_LDH,

ME and

_SOD,

and a

Tris/citrate

_system,

_{pH 6.2,}

(Nichols

and

_Ruddle,

₁₉₇₃₎

for GAPD. For PGM and

_TPI,

both buffer

_systems

were used. After

_{electrophoresis}

(5

_h,

maximum 30 mA or 300

_V/gel),

_gels

were

stained for enzyme

activity

according

to Harris and

_Hopkinson

_(1978).

The

nomen-clature for the buffalo _{enzymes is} based on the _concept of

_homology.

Abbreviations

and criteria for

_homology

are in

_agreement

with the ISGN

_system

_(Human

Gene

Mapping

5,

1979).

Zymograms

are

presented

in

figure

1.

Buffalo tissues and hamster cells showed one zone of GAPD

_activity,

with

different

_mobility.

The 3 intermediate zones in

_{hybrids expressing}

buffalo GAPD

LDH is a tetrameric _enzyme coded for

by

two

_separate

_genes, LDHA and LDHB.

In buffalo

_tissues,

5

_isozymes

_{(LDH-1-LDH-5)}

could be

_{distinguished.}

These may

be considered as

being

composed

of different combinations of the A and B

subunits,

subunit A

_being

the main subunit in skeletal muscle

_(M)

and subunit B

_being

the

main subunit in cardiac muscle

_(H)

and

erythrocytes

(RBC).

In the

_hybrid

_panel,

all 4 combinations of _presence or absence of buffalo LDHA and LDHB were found.

Two forms of

_ME,

the

_cytosolic

_enzyme ME1 and the mitochondrial enzyme

1975).

From the relative

_activity

of the two ME zones in buffalo

_tissues,

and

by comparison

with the relative

_mobility

as found for bovine ME

_(Heuertz

and

Hors-Cayla,

1981),

it was concluded that the most anodal zone _represents buffalo

1VIE2,

and the more cathodal zone buffalo MEI. In

_{hybrids expressing}

buffalo

_ME1,

intermediate zones could be detected. Buffalo ME2 was not found in _any of the

hybrid

cell lines.

In _man, 3 _genes are described for PGM

_(Harris

and

_Hopkinson,

_1978).

The relative

_migration

order of bovine _{enzymes is} the same as that of human _enzymes.,

ie,

from cathode to anode:

PGM1,

PGM2 and PGM3

_(Heuertz

and

_Hors-Cayla,

1981).

The zone with the lowest

_mobility

in buffalo tissues was also found in some

of the

_hybrids.

From its relative

_activity

and relative

_mobility,

it was concluded that this zone _represents buffalo PGM1. The absence of intermediate zones in

_hybrids

expressing

buffalo PGM1 indicates that this _{enzyme is} a monomer.

In buffalo

_tissues,

two

_separate

zones of SOD

_activity

were found. From the

absence of the cathodal zone in

_erythrocytes

_(RBC),

and

_{by comparison}

with

zymograms of other

species

(van

Someren et

_al,

_1974;

McAvin et

_al,

_1988),

it

was concluded that the anodal zones _represent the

_cytosolic

_enzyme SOD1 and the cathodal zone the mitochondrial _enzyme SOD2. The one intermediate zone

in

_hybrids

_expressing

buffalo SOD1 indicates a dimeric structure for this _enzyme. Human SOD2 has been proven to be a tetramer

_(Harris

and

_Hopkinson,

_1978).

No buffalo SOD2

_activity

could be detected in any of the

_hybrids

at the site

corresponding

to SOD2 in buffalo tissues. The conclusion that buffalo SOD2 is

expressed

in

_hybrids

is based on the _presence of one minor intermediate zone, and

the

_supposition

that hamster SOD2 is far more active than buffalo SOD2.

Human TPI1 is a dimeric _enzyme

_present

in all tissues

_(Harris

and

_Hopkinson,

1978).

Many

secondary

isozymes

can be

detected;

their number varies with the

_type

of tissue and the buffer

system

used for

_separation.

In the buffalo tissues as well as

in the hamster and

_hybrid

_cells,

_secondary

_isozymes

of minor

_activity

were found

(not shown).

In buffalo tissues and hamster

cells,

one

_major

zone of

_activity

was

detected. The one intermediate zone in

_{hybrids expressing}

buffalo TPI1 indicates a dimeric structure for this enzyme.

The

_segregation

_pattern

of buffalo genes in the

panel

of

_hybrid

cell lines

investigated

is

_given

in table I.

_Independence

of _presence or absence of two _genes

was

_{analyzed using}

the

X

2

test with Yates’ correction for 2 x

_{2 contingency}

tables.

Synteny

was concluded when P < 0.01.

_By

this

_criterion,

_GAPD,

_{LDHB, TPI1,}

ME1 and SOD2 can be considered

_syntenic.

_LDHA,

PGM1 and SOD1

_segregate

independently

from each other and from the two

_syntenic

_groups

_(P

>

_0.01),

and

are

probably

located on _separate chromosomes.

The _genes found to be

_syntenic

in the buffalo are also

_syntenic

in Bos taurus cattle

(Fries

et

_al,

_1989),

and in man

(Human

Gene

_Mapping

_10,

_1989).

This indicates conservation of

_genomic

_segments

in these 3

_species.

In the _mouse,

_synteny

exists

for

_GAPD,

LDHB and

_TPI1,

but not for ME1 and SOD2

_(Mouse

_Genome,

_1990).

The 3 genes found to

_segregate

_{independently}

in the buffalo

_belong

to

_independent

groups in the 3 other

species

as well.

REFERENCES

Echard

G,

Gellin J, Gillois M

₍₁₉₈₄₎

Localisation des

_genes

MPI, PKM2, NP sur le chromosome 3 du _porc

_(Sus

_scrofa

_L)

et

_{analyse cytog6n6tique}

d’une

_lign6e

de hamster

chinois issue de la DON

_(wg3h).

Genet S61

Evol 16,

261-270

Fries R, Beckmann

_{JS, Georges}

M, Soller

M,

Womack J

₍₁₉₈₉₎

The bovine gene map.

Anim G6n6t 20, 3-29

Harris H,

Hopkinson

DA

₍₁₉₇₈₎

Handbook of _enzyme

_{electrophoresis}

in human

_genetics.

North Holland Publ

_Co,

Amsterdam

Heuertz

_{S, I-Iors-Cayla}

MC

₍₁₉₈₁₎

Cattle gene

mapping by

somatic cell

_{hybridization:}

study

of 17 enzyme markers.

_Cytogenet

Cell Genet

_30,

137-145

Human Gene

_Mapping

5

₍₁₉₇₉₎

International

_System

for Human Gene Nomenclature.

Cytogenet

Cell Genet 25, 96-116

Human Gene

_Mapping

10

₍₁₉₈₉₎

Tenth International

Workshop

on Human Gene

_Mapping.

Cytogenet

Cell Genet 51, 1-1148

lVIcAvin

_JC,

Patterson D, Womack JE

_{(1988) Mapping}

of bovine PRGS and PAIS _genes

in

_hybrid

somatic cells:

_syntenic

conservation with human chromosome 21. Biochem Genet

26, 9-18

Meera Khan P

₍₁₉₇₁₎

_Enzyme

_{electrophoresis}

on cellulose acetate

_gel:

_{zymogram patterns}

in man-mouse and man-Chinese hamster somatic cell

_hybrids.

Arch Biochem

_Biophys

145,

470-483

Nichols

_EA,

Ruddle FH

₍₁₉₇₃₎

A review

_{of enzyme polymorphism, linkage}

and

electropho-retic conditions for mouse and somatic cell

_hybrids

in starch

_gels.

J Histochem

_Cytochem

21, 1066-1081

Povey S,

Wilson

_DE,

Harris

_{H, Gormley IP, Perry P,}

Buckton KE

₍₁₉₇₅₎

Subunit structure

of soluble and mitochondrial malic enzyme: demonstration of human mitochondrial enzyme in human-mouse

hybrids.

Hum Genet London 39, 203-212

Shows TB, Tashian

_RE,

Brewer GJ

₍₁₉₆₄₎

_{Erythrocyte glucose-6-phosphate}

dehydroge-nase in Caucasians: new inherited variant. Science _145, 1056-1057

van Someren

_{H, Beijersbergen}

van

_{Henegouwen H,}

Los

_W,

Wurzer-Figurelli E, Doppert

B,

Vervloet

_M,

Meera Khan P

₍₁₉₇₄₎

_{Enzyme electrophoresis}

on cellulose acetate

_gel.

II.

_Zymogram

_patterns in man-Chinese hamster somatic cell

_hybrids.

_Humangenetik

25,