Analysis of isoenzymes in trophoblast cells

Cell Biology International Reports, VoL 10, No. 3, March 1986 ANALYSIS OF ISOENZYMES IN TROPHOBLAST G.Novelli,F.Catizone,B.Dallapiccola.Department Universities The expression fetal

of Urbino and Rome

of proteins

development

development diagnosis requires

of Rossano

in early human

use in monitoring

at risk for genetic

metabelic

u

pre-

mm

dim _ _

seases. We have determined of glycolytic

the isoenzymatic and dehydrogenase

in 6 trophoblast

samples

collected

8 and 12 weeks of gestation

ween

m

patenz Z

m m

mm m m

--

mm

m

m =m

m

mes

Calabro,Italy

of first trimester fetal this information in vi-

ew of its potential

terns

CELLS of Genetics,

is still poorly understo

od.The

gnancies

II and Hospital

149

m

mm

m

mm

bet

m m

in women

requesting legal abortion. Chorionic villi were selected under an inverted microscope to avoid contamination of tissue of maternal origin.Fetal origin of examined cells was established by proving absence of GLO !I and ALDH~.Cell extracts were processed I as previously described .Phenotypes were examined

on cellulose

acetate

GPI

pGM

ALP

LOH

G6PD 6PGD

ICD

MDH

ADH

GDH

and starch-

gel electrophoresis according to the methods of Hat{is and Hopkinson and van Someten et al. We have shown that: enzymes glucose

HK

Fig.l-

Diagram

showing

approximate

ve mobilities isoenzymes examined phoretic systems not identical)

relati(electro

1.Both HK and HK iso i 2 of hexokinase are present; 2.The phosphate isomerase isoenzyme co-

ded by a locus on chromosome !9 is expressed; 3.The phosphoglucomutase isoenzymes, PGM PGM and PGM are present" a. There a-

1'

2

3'

'

re three aldolase bands,possibly c o r r e s p o ~ dinE to the A and C loci and their respective heteromeric

combinations;

ement with observations eues,trophoblast

5. In agre-

in other

ceils express

fetal

tis

the five ma

lactate dehydrogenase isoenzymes (LDH I ); 6.Glucose-6-phosphate dehydrogena5 se and 6-Phosphogluconate dehydrogenase a-

observed; rms

8. Not l e s s

of malate

than four anedic

dehydrogenase

jor

which are derived

-LDH

MDHm

re present with single bands; 7.Only the ICDs isoenzyme of citrate dehydrogenase is

tide subunit of alcohol dehydrogenase is present; i0. Glucose dehydrogenase is pr~ sent as a single band (Fig.l).

types;

both

fo

are expressed,

from the MDHs

9. Only the ADHlalpha

and

polype R

REFERENCES 1.

Oallapiccola,B.,Novelli,G.,Palloni,R.,Catizone,Y.,Magnani,M.in:-First Trimester Fetal Diagnosis ( F r a c c a r o , M . , S i m o n i , G . , B r a m b a t i , B . , e d s . ) . S p r i n g e r , B e r l i n 1985,pp.

2.

Harrls,R.,Hopkinson,D.A.in:-Handbook of Enzyme Elsevier-North Holland,Amsterdam,Z976.

3.

van Someren,H.,van

2a2-245.

M.P.(I974)

Electrophoresis

in Human

Genetics.

Henegouwen,H.B.,Wurzer-Figurelii,E.,Doppert,B.,Vervioe~,M.,Khan,

Humangenetik 25,!89-201.

ACKNOWLEDGEMENTS This work was partly supported by a grant from Progetto Yinalizzato Ingegneria tica e Basi Molecolarl Malattie E r e d i t a r i e , I t a l i a n CNR,con~raet N.SZO140a-51

0309-1651/86/030149-01/$03.00/0

Gene-

© 1986 Academic Press Inc. (London) Ltd