Karyotyping prometaphase chromosomes of alpaca (Lama Pacos, fam. Camelidae)

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CENTRE OF GENETICS _{AND BIOTECHNOLOGY}

oF THE ul'ilvERslrY oD TRAS-os-ritoNTES AND ALTO DouRo

17th

_{EUROPEAN COLLOQUIUM ON}

_ANIMAL

CYTOGENETICS

_AND

_{GENE MAPPING}

Book

_of

_AbsÉraeÉs

18-21June

2006

Auditorium of

calouste

_{Gulbenkian Foundation,}

§ene tuapVing -

1S-Uj::,

20C6 _ /_ts|on,

?r:y:{

_-

_---PCSTERS

_SESSIOI.."r

"Gene

Mapping

_{qnd Chromosome Maps,,}

P1

Cytogenetic localizati on of

FtVRl

gene

_in

farm animals

Danielak-Czech _{B, Slota E}

Deparftnent of Animal Immuno_and Cytogenetics, National Research Institute of Aaimal

_ilrùr"tior,

Balice/IGakdw, poland

We have _{localized trinucleotide repeats}

_of

_the FRMI gene in cattle, sheep and _{pigs karyospes}

using in siru pCR method with

p;i".,

_fl-i,i"g

CCG repeats located

_in

_the

_5,LffR

_regulator!

regron of the human FMR1 mRNA.

The

GCTCAGCTCCGTTTCGGT'ff

_CAC##J§'

T

(forward)

and

AGCCCCGCACTTCCACCACCAGCTCCTCC

,A (reverse) were used

for

in

situ pCR

and

biotin-16 dUTp labeling

of

CGG containing

ly!

gene _{fiagment (1}

_cycle:

_{94. C _ 3}

_.j",

6t

C

-

1

yi\72'

C

-

I

mìn; 30 cycles: 94" C

_

I

min, 65'C

_-

I

min, 72. C

_

I

_{m;n) on eFe_}

and

GTG-banded'metaphase

"Éo-o*",

1lides. Fluorescent ampliÈcation signals were detected

by

_{avidin-conjugated}

_ÉfC

.and mapped on cattlg sheep and pig

X

chromosome

at BTA

Xp13,

_{OAR XqZZ}

_{and SSC Xq26}

regions, respectively.

In

conclusior, _{the results obtained suggest that}

there _{are similarities of regulatory seqGnces} in CGG-region of FRML gene in

aif"r"ot

species.

The identified locus-specific sequenc"

_*,

U"

gflird

in _{comparative and evolùtionary}

studies. This work was conducted _as

_part

_oi

_NRLAP statutory activity, project _{no- 3210.1}

P2

Karyofyping

_{prometaphase chromosomes}

of alpaca (Lama_pacos,

_family

_Camelidae) Di Berordino Dt, Nicodemo DI, Fauciutto

el,

'

Cosenza Gt,. Ramunno Lt, King AW.. Coppota

G',

Irn*rrri

il

,

ni u"o

Ci-r,"iri*;Zrt

Rubes J6

rDepartneat

ofAlimal Science uo6 pqed rnspection, Universiry of Naples .Tederico _II,,

Portici,Naples,Italy; 2Department _{of Biomedical}

i:fu1.o,

Universiry of Guelph,Ontario, Cauada;

Alational Research Couucil (CNR), ISPAAM

Laboratory ofAnimal _{Cytogenefics and Gene}

Mapping, Naples, Italy; aDeparment _{of Clinical} Vrcterimry Medicine, _{Centre for Veterinary,science,} University of Cambridge. Cambridge, IJK]5 Eacutty of Veterinary Medicing _{Iasi, Romania;fueterinary} -Research Institute, Brno _{, Czech Republic}

$-

k"o*O

the alpaca (Lama pacos,2n:74.Xy)

belongs

to

the

family

Camelidae, suborder

Tylopoda, which includes the vicugna (Lama

vicugna),llama (Lama glama), grruù.o _@amo guanicoe)

_and

_{the two}

_{camels, bactian} (Camelus bactrianus) _{and dromedary (Camelus}

dromedarius)- Despite the increasing worldwide

economical

_interest

_{in tht}

_peculiar

characteristics

of

_its

_wool

_(extemely

_fne, unallergic

and

naturally _{stained), the'alpaca} species has been

_-so

far- almost forgotten from

a

cltogenetic

_point

_of

_view.

_[r

_order

to

contribute

_{to the}

_{clarification}

_of

_the

chromosomal constitution

of

this

species, we

decided

to

undertake

_a

_{derailed iytogenetic}

fves{rylo1

thus providing, for

the'trsi

,t-,

the

_{QFQ-C-GTG-RBG}

"rd

RBA

banded

karyotypes _{and the GTG_RBG ideograms,}

at a

resolution

of

_nearly

₄₀₀

_bands. -Based _on centromeric

index

and

relative chromosome

length measurements,

_the

_alpaca

_k

_ry"qrp;;

been arranged

_in

_{four goups}

_of

_{chromàiom"s,} as follows: _{goup A, subtelocentrics,}

_{fronfair I}

to 10; group _{B, telocentrics, from par}

_fi

_[o

20;

group C, submetacenkics,

_from

_pau

_2l

_to

_29; group _{D, metacentr.ics, from pair3O}

to 36. Six

paim of nucleolar _{organizer c-hro-oro-".- hur"} Deen

_{also identified}

_by

_using

_a

sequential

RBA./silver _{steinjlg technique}

_uJ".

O

fa}orpl,

28

(c

group)

_,3t,

_i2,33

;d

₃₄

@

grì"ii.

m,

j

present karyorypes _{and the GTG/RBG_banded}

ideograms can be- used for clini€1 cytogenetics,

karyotype

standardization, _{gene- mapping,}

:ytotaxgnomy, comparative studies ana

gìn"ti"

tmprovement prograrns

_within

_the

_{a_ify}

Camelidae.

P3

Chromosomal _{mapping of ribosomal}

genes _{in venerid clams}

@ivalvia: Veneridae)

Hurtado _{NS, Iv{ordn} p. pasantes

_JJ

D_pto Bioquimic4 _{Xenéfica e Inmqletez64} Universidade de Vigo, _{E-363 lO Vig;;}

_$irn

The- fàmily Veneridae

_{is a}

_group

_of

_bivalve molluscs found in all the

*o.Ia

,*-

Although th9 famity _{includes some 5@ qreciw,}

Àrny

of

which are

of

conamercial

_àprr*ì"

.and

rydgcted to intensive exploitatioi

"o

Lo

oo

-cnro3osog3t mapping

oi

oNe

*"d"; ;;

specles of Veneridae was _{reported to}

date-In

order to localize both the major

_*J',U.

_SS

P-NA

gene

clusters," fluoreslcent

_in

_situ

hybridization

_(FISIù

_experiments

were

performed on mitotic chromos-omes _{and surface}

Tr"ud

synaptonemal complexes of the venerid

clams

potlria

_exollta,

_{i"ait"po i"*rror^,}

Ruditapes

_philtipinarum

and

i.i"*pt"

pullastra.

_Species

_specific

_p.ob"s

*"..

generated

by

pCR amplificatioi

_of

_{both the} intemal transcribed _{qpacers and the}

5.g rDNA

of

the

majol

_r?NA

_{gene cluster ana}

_tle

wUo1e repeat _{unit of the 55 rDNA cluster.}

The

repeats

_of

_the

_major

_rDNA

_genes

are clustered _{in only one cbrohoso-"}

_priiio

uU

t"

species analyzed, the NOR is termàal

_l"

*r;;

the species and intercalary

_i,

_A"

_otn.rLo.'fn"

also unique 5S rDNA _{gene cluster}

_,rr"*

_

intercalary

_or

_terminJ

_{_ in ;}

-àiÀ.o.o,

chroSoyme

_pair

_in

_three

_of

_the

_fo*

"t"*

species _{but co-localizes with the majoruONa} cluster

_n

Ruditapes _{decussalw. The co_location}

was confirmed _{by FISH on extended chromatin} IIDTES.

P4

Recurent

_airway

_obstruction

_(RAO)

in

horse: _{chromosomal an}

_RII

_*rpf#g

of

three candidate genes

Nfknlqska Ròtzter _-/,^Bugno

_t?,

_Sloto

_{È, Ooy}

9,

th.ypary

BP,

s*iinurn"tÉ,;;;

b

-Gerber

I

]P:rg*:*

of Veterinary Ctiaical Studies,

,VetsuisseFacul-ry_Universityof

r"_à,iJitorura

uepartutent of _{Immrmo and Cytogenetics, National} |eJearch Insrirure of Animal

p.Lar'.i1",É';;'

Poland

3Institute

of _{Genetics, Vetsuisse Faculty,}

University

ofBerne, Switzerland ' Departuent of Veterinary_

_Integrative Biosciences, Texas _{A&M Uuiversiry, USA}

'Animal _{Health Trusg Newmarket,}

Suffolh

_IIK

Ì.

I""uo."ll.

lhuy

obstruction

(RAO)

_of

the

norse rs

lftely

a _{polygenetic disorder and shares} many characteristic _{features with human}

asrhrna

and

cbronic

_{obstructive putroo".y-disease}

(COPD). _{RAO and asthma}

_*

"nu*É"rlr"a

Uy

:T^y^^

-tfonchospasnq

coughiag

ut

*uy

nyperreachvity,

_inflammation

_and

mucus

accumulation. Select genes:

_EGF&

_CLCAI, BLC2 arc of interest in rÈspiratory _airo.a".,

"1m

chronic mucus

ove.pioduction,

_i""i"a.g

asthma_

_In

_{our study, we}

-upp"à

,n".è

fr*

genes

_by

_FISH

_on

_equine

-"Lpnu."

spreads.

Equine

BAC

librarys (CHORI,

-ÀIRi)''*"r.

screened _{with equine gene_specific}

primérs and

positive clones _{were iabelleà with}

_{àffig.r_}

and

then

used

_for

_FISH

_on

_{Ciè-_'U-*a.O}

chromosomes.

_{The BAC}

_clones -

narU"ìr.g

these three genes were mapped _{to the following}

chromosomes:

_{EGFR_}

_{Écxpti,'-Cicer}

ECA5q15, BLC2_ECA8qzz.

_aJÀu-oouii.

_*"

confirmed the _{results with the nff__upplilg}

oo the _{TAMU radiation hybrid}

_{panef-fn"'J"iirg}

data, _{which confirm}

_tlé

_estaUnsn"a

"oorl#utioo

o.f

_{synteny between equine}

_and

nr_u,

chromosomes, _provide

_a

_resource

for

further

association studies

_of

_these

_g"r.,

_i,

"qìir.

RAO.

f7,