European Journal of Zoological Research
Abstract
Author(s): Forutan Salehi Nezhad, Akram Yar Ahmadi and Zahra Ghandali Varnaseri
Testis is an organ that its natural function depends on temperature. It was determined that heat stress in male
reproductive system followed by at leasttwo negative results. One of them is the exposure of oxidative reaction and
the other one is impairment in DNA structure. Thus the present study was conducted aiming at evaluating oxidative
reaction indicators and the exposure of DNA damages by using TUNEL method in Sertoli cells under heat stress at
42 in order to determine how heat stress can affect Sertoli cells and study the sort of resulted damages in these
cells under these conditions, for this testes of 9 male lambs of 3-10 months were provided from slaughter house and
the separation and culture of Sertoli cells were performed on these samples. To evaluate heat stress effect, Petry
Dishes containing Sertoli cells were divided into 3 categories; first category was control group and was incubated
at 32 . Second category which were under mild thermal stress at 39 centigrade for 6 hours and the third category
were under severe thermal stress at 42 centigrade for 6 hours. In order to account cells which are involved in
planned death, colored TUNEL was done on three positive, negative control group and test. Four parameters
related to oxidative reaction in control group and affected groups under thermal stress were conducted in peroxidation-
lipids test via measuring MDA concentration (TBARS),measuring the activity of super-oxide-dismothase
enzyme, measuring nitric oxide quantity and FRAP test to study total value of oxidation compounds in the culture
medium of cell. Finally DNA damages and oxidative reaction occurrence in control groups, under mild heat stress
and under severe heat stress, were studied by experimental design (Random design) and statistical test (One way
ANOVA) in statistical software (SPSS). The results of present study showed that by increasing temperature the rate
of living cells decreases and the difference between living cells percent in control group 32 centigrade and the
groups of 39 and 42 centigrade is significant (P<0.05). TBARS rate increases from control group to higher
temperature and there is a significant difference between control group and the group of 42 centigrade (P<0.05). by
increasing the temperature the amount of nitric oxide increases. More specifically, there is a significant difference
between control group and the group of 42 centigrade (P<0.05). TUNEL dying evaluating showed that applying
thermal stress causes an increase in the number of damaged cells from DNA and this increase in the group of 42
centigrade is more significant than control group (P<0.05).
