J Neural Transm (1990) [Suppl] 32: 161-164

by Springer-Verlag 1990

Monoamine oxidase activity in blood platelets of

migraine patients

M. M. Caramona 1, M. D. Cotrim 1, C. Fontes Ribeiro 2 , and T. Macedo 2

1 Laboratorio de Farmacologia, Faculdade de Farmacia, 2 Instituto de

Farmacologia e Terapeutica Experimental, Faculdade de Medicina, Coimbra,
Portugal

Summary. Biochemical changes in platelets of migraine patients during the

attacks have been reported before, however there are some conflicting
results. In an attempt to define the biochemical lesion in the platelets, we
have carried out a survey of platelets monoamine oxidase in migraine
patients with and without aura. Platelet MAO activity in platelets from
migraine patients was significantly reduced when compared with normal
platelets.

Introduction

Monoamine Oxidase (MAO) plays an important role in the oxidative

deamination of a wide variety of biogenic amines like catecholamines and
serotonin. Based on substrate specificities and susceptibilities towards in-
hibitors, MAO can be divided in two forms, namely MAO-A and MAO-B.
In human platelets the MAO is present in the MAO-B type (Donnelly and
Murphy, 1977).
Interest in MAO activity in migraine patients was stimulated by the
observation that the "cheese reaction", (the hypertensive reaction to certain
foods in patients receiving MAO inhibitors), was similar to the dietary
migraine episode; as this reaction is thought to be due to tyramine, it was
suggested that patients with dietary migraine might be sensitive to tyramine-
containing foods by virtue of a deficiency of MAO (Hanington and Harper,
1968).
Platelet MAO activity has been found to be significantly decreased
during migraine attacks when compared with activity in the crises-free
period (Glover et aI., 1977, 1982). It was also found that mean platelet
MAO activity between attacks in males with classical migraine, with tension
headaches and with cluster headaches was significantly lower than that in
162 M. M. Caramona et al.

a control group of normal men (Glover et aI., 1981). However, conflicting

results have been reported possibly due to differences in technical proced-
ures used or to the heterogenity of the migraine patients (Thomas, 1982).
Having in mind the headache classification of the National Institute of
Neurological Diseases and Blindness (USA-1988) and in an attempt to
define more clearly the biochemical lesion of migraine, we have carried out a
survey of platelet MAO activity in migraine patients with and without aura
using only platelet samples from subjects in interval periods, i.e. between
attacks.

Materials and methods

The migraine patients were selected from a group of females and males presenting
themselves without an attack at Coimbra University Hospital (Headache Group
of Neurology Service). The subgroups with and without aura were classified on
the basis of the criteria of the National Institute of Neurological Diseases and
Blindness (USA -1988).
The normal subjects belong to the Blood Donnors Service from the same
hospital and all of them were healthy volunteers.
All the subjects were in diet (phenylethylamine and tyramine absent), without
medication for at least 14 days and not depressed (Back Scale). The migraine
patients had not experienced an attack during at least the last five days.
The platelets were prepared using a modificated method described by Fowler
et al. (1979). Twenty ml blood samples were obtained by venipuncture and collected
in standard heparine/lithium tubes; the subjects were aged between 20 and 60 years
and the blood obtained, in the morning (9-10 a.m.). After being gently mixed, the
blood was centrifuged at 200 g for 10 min. to remove the red cells. The cells were
washed twice with 0.15 M NaCI and the combined supernatants further centrifuged
at 3,000 g for 15 min. to produce a platelet preparation. The pellet was resuspended
in 2 ml of 0.01 M potassium phosphate (pH 7.4) and stored at - 80C. Before use
the suspension was sonicated in a Tissumizer Ultrasonic Desintegrator at low
power for 2 min., to produce a more homogeneous preparation.
Monoamine Oxidase Assay: MAO activity was determined using 14C-Phenyl-
ethylamine HCI (PEA, 50 mCi/mmole, New England Nuclear) as a preferential
substrate for MAO type B. The reaction mixture containing 50 ~l platelets suspen-
sion and 50 ~l of 0.01 phosphate buffer (pH 7.4) with the substrate was incubated
during 20 min.; the de aminated product was extracted and measured by liquid
scintillation counting (Caramona, 1982).
To obtaine information about the Km and V max in the human platelets the PEA
concentration~ used were 2, 4,8, 16,32, 64 ~M. The individual measurements were
performed with PEA 100 ~M. MAO activity is expressed in nanomoles of substrate
metabolized per milligram of protein per hour of incubation.
The protein content of the platelet suspensions (milligram of protein per ml of
suspension) was determined by the method of Lowry et al. (1951) with bovine serum
albumin as a standard.

Results

The rates of deamination in Dlatelets from controls and in those of migraine