L. Marchat et al., ISOENZYMATIC DIAGNOSIS OF FILARIAE - A METHOD FOR SEPARATION OF LACTATE-DEHYDROGENASE ISOENZYMES FROM MOLINEMA-DESSETAE (NEMATODA, FILARIOIDEA), Comparative biochemistry and physiology. B. Comparative biochemistry, 109(2-3), 1994, pp. 451-457
Lactate dehydrogenase (LDH) is highly active in filariae and could be
a valuable tool for phyllogeny studies. Unfortunately, the isoenzymati
c diagnosis of filariae is often difficult for LDH because of a poor m
obility of the enzymes in starch gels which are the most commonly used
in such studies. We propose here a method to separate filarial LDH is
oenzymes using disc electrophoresis. The experiments were carried out
on male and female Molinema dessetae in order to compare their respect
ive isoenzymes. The study of several parameters such as buffer systems
, percentage of bisacrylamide and progression time led to optimize the
enzyme separation. LDH from male and female filariae were compared to
mammal LDH-H4 and LDH-M4. Five and four LDH isoenzymes were found, re
spectively, in male and female worms. Relative concentration of each i
soenzyme diverged between male and female worms. Mammal muscle LDH-M4
type moved between LDH2 and LDH3 from female worms, and between LDH1 a
nd LDH2 from male worms. Mammal heart H4 type enzyme was very differen
t in electrophoretic mobility. The ratio of each isoenzyme was determi
ned by densitometry. The major isoenzymes from female worms will be st
udied as a biochemical target for chemotherapeutic attack.