OPTIMIZATION OF COUNTING PROCESS OF DOPAMINERGIC NEURONS IN SUBSTANTIA NIGRA OF PARKINSONIAN MICE
V. G. Khaindrava,1,* P. V. Ershov,1 V. E. Anciperov,2 J. V. Obukhov,2
A. K. Nanaev,3 M. V. Ugrumov 3
1 P. K. Anokhin Institute of Normal Physiology RAMS, 2 Institute of Radiotechnics and Electronics RAS
and 3 N. K. Koltzov Institute of Developmental Biology RAS, Moscow;
* e-mail: khaindrava82@mail.ru
Parkinson’s disease (PD) results from degeneration of dopaminergic (DA-ergic) neurons of sunstantia nigra
pars compacta (SNc). In experimental studies, this condition is modelled by administration of neurotoxin’s precursor
1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). The followed degeneration of DA-ergic neurons
is estimated by cell counting. Cell counting with serial sections is exceedingly hard and expensive. Therefore
this method is not used extensively in spite of its high precision. Widely known Konigsmark’s formula
(KF) allows not to count cells in all serial section, but only in sections selected at regular interval. However, its
precision decreases with increasing the interval and other parameters. In given work we have developed mathematical
method of approximation (MA) by improving of KF. MA allows reducing the time and chemical products
expenses for processing and analysis of the material with maintenance of necessary counting precision.
Two groups of C56/BL mice were used in this study. The first group received MPTP in a dose of 8 mg/kg with 1
week interval and showed 20 % decrease in DA-ergic neurons in SNc. The second group received MPTP in a
dose of 12 mg/kg four times with 2-h interval and showed 40 % decrease in DA-ergic neurons in SNc. Degeneration
took place mostly in the middle part of SNc within rostra-caudal direction and led to rise of sharp differences
in the number of neurons from section to section. These differences substantially decreased precision of
FK (6 % error with counting in every 5th section), whereas precision of MA was sufficiently good (4 % error
with counting in every 7th section). Thus, we have developed MA, that allows us to decrease expenses of the time
and chemicals for estimation of DA-ergic cells number in SNc of parkinsonian mice.
Key words: Konigsmark’s formula, substantia nigra, dopaminergic neurons, neurotoxin, mathematical analysis, parkinsonism, tyrosinhydroxilase, mice
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