Biology Reference
In-Depth Information
26.2
Methods
26.2.1
Chemicals and Reagents
As
2
O
3
, NE, DA, and 5-HT were purchased from Sigma Chemical Company
(St. Louis, USA). HPLC-grade methanol was supplied by Sinopharm Chemical
Reagent Co., Ltd. Analytical grade sodium acetate was bought from Shanghai
Chemical Agent Cooperation (Shanghai, China). PrimeScriptTM RT reagent Kit
and SYBR
®
Premix Ex TaqTM kit were supplied by TaKaRa (Dalian, China). Other
reagents were commercially available.
26.2.2
Animals and Treatment
Sixty SPF mice (7 weeks old, 26.3-30.9 g) were supplied by Experimental Animal
Center, Dalian Medical University in China. The animals were caged under a 12 h
dark-light cycle in standard conditions of temperature (18-22°C) and humidity
(50%). The mice were randomly divided into three groups after 7 days acclimatiza-
tion. Groups 1 and 2 were administrated with drinking water alone as control or
4 ppm As
2
O
3
ad libitum for 60 consecutive days, respectively. The protective group
was treated with both 4 ppm As
2
O
3
and 150 mg/kg taurine. The protocol was per-
formed in accordance with the Animal Guideline of Dalian Medical University and
in agreement with the Ethical Committee of Dalian Medical University.
26.2.3
Ultrastructure of Synapses
For the synapses ultrastructure, brain tissues of mice treated with drugs were pre-
pared after the final treatment. Then the tissues were immediately fixed in a 3%
glutaraldehyde solution in PBS (pH 7.2) overnight at 4°C and washed in 0.15 M
PBS for 30 min. Then the samples were postfixed for 2 h in 0.1 M PBS osmium
tetroxide (1%) solution at 4°C, dehydrated in graded EtOH, and embedded in
Araldite (Fluka, Buchs, Switzerland). The ultrathin sections were contrasted using
both lead citrate plus uranyl acetate. And the samples were observed by transmis-
sion electron microscopy (TEM) ( JEM-2000EX, Olympus, Japan).
26.2.4
Quanti fi cation of Monoamine Neurotransmitter
Levels by HPLC
The brains of mice in the experiment were removed and stored at −80°C after
the final treatment. Then the tissues were weighed before homogenizing with
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