Physiological saline of Control animals

Experiments were carried out on male Wistar rats.
Experimental DM1 was induced by single streptozotocin administration (65 mg/kg, intraperitoneally) to
3-month old rats. Control animals received physiological saline (0.9% NaCl pH 4.5). DM1 development in
rats was associated with pronounced hyperglycemia
(22.1±1.0 mmol/liter), persistent glucosuria (28-56
mmol/liter) and polyuria. Neonatal DM2 was induced
by streptozotocin administration (80 mg/kg) to 5-day
old rat pups [6]. In 2.5-3 months, DM2 sings appeared
detected using glucose tolerance test. Only rats with
glycemia 1.5-2-fold surpassing the normal after glucose load test (2 mg/kg) were included in the study.
Marked glucosuria (14-28 mmol/liter) was also noted
in rats with DM2.
To obtain heterogeneous cell membrane fraction
from rat brains, the cortex was homogenized in 1mM
NaHCO3
buffer pH 7.5. Homogenate was centrifuged
at 8000g for 20 min (4o
C). Supernatant was diluted in
buffer and centrifuged at 80,000g for 30 min (4o
C).
Precipitate was resuspended and used in subsequent
experiments. Protein concentration was measured by
the method of Bradford; γ-globulin was used for construction of the calibration curve.
Specifi c binding of 125I-insulin by rat brain cell
membranes was assayed by radioligand method with
plotting the curves of competitive displacement of
receptor-bind labeled insulin by unlabeled hormone.
The brain cell membrane fraction was incubated with
125I-insulin in the presence of 0.1-1000 ng/ml of unlabeled hormone. Non-specifi c binding was determined
by adding 10 μg/ml unlabeled insulin. Data of radioligand analysis was transformed into Scatchard coordinates to calculate KD and RО that characterize affi nity
and number of binding sites, respectively.
Expression of IRS2 protein gene was d


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