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Separatration of early stage acrosome reacted sperm and analyses
of the proteins
Kenji Yamatoya, Chizuru Ito, Cheng Chen, Mamiko Maekawa,
Yoshiro Toyama, Kiyotaka Toshimori
Dept. Anatomy and Developmental Biology, Grad. Sch. Med., Univ. Chiba
During the acrosome reaction, the contents of sperm acrosome are released from
the original location to the final destination and working place. The molecular
distribution and nature are posttranslationally modified. We reported that flow
cytometric analyses using antiacrosomal membrane protein antibodies anti
IZUMO1 and antiSPACA1 can discriminate the early stage of acrosome reaction,
classifying into 7 populations.
In this study, we analyzed the 7 populations and found that the populations
number 4 and 5 contain acrosomeswelling sperm. Acrosomeswelling occurs at
a very early stage of acrosome reaction which starts before the exposure of inner
acrosomal membrane. One of the acrosomal membrane proteins, SPACA1, was
cleaved during this swelling stage. Western blot showed that SPACA1 was 35 kDa
before acrosome reaction and became 21 kDa in acrosomeswelling sperm.
Thus, this flow cytometric method is useful to analyze the detailed acrosome
reaction process and the molecular changes during the acrosome reaction which
are the last preparatory process for the spermegg interaction.
P
Cell adhesion molecule Nectin
Kannika Adthapanyawanich
Cell adhesion molecule1 Cadm1 Nectin3
Cadm1
Poliovirus receptor
Nectin3 Nectin2
Cadm1 Nectin3
Cadm1 Nectin3
Cadm1//Nectin3/ Cadm1/
Nectin3/
Cadm1 Nectin3
P
SF
SF1
2 anti
Müllerian hormone type 2 receptor Cre CreloxP
SF1
0 21 SF1
7
14 21
14 21 SF1
AMH
p27
WT1SOX9GATA4androgen receptor
SF1
SF1
P
Autoimmune responses induced by immunization with xenogenic
testicular germ cells alone
Experimental autoimmune orchitis EAO is one of the models of immunological
male infertility. Classically, the immunization of mice with a testicular
homogenate emulsified in complete Freund's adjuvant followed by intravenous
injections of Bordetella pertussis is necessary for the induction of murine EAO.
Later, we previously established a mouse EAO model that can be induced by two
subcutaneous injections of viable syngeneic testicular germ cells TGC alone,
and the autoantigens involved in the mouse EAO have been analyzed using two
dimensional gel systems and western blotting. In the present study, to examine
whether the xenogenic TGC can induce a mouse EAO, we immunized mice with
viable TGC taken from the rat or the guinea pig. The results showed that mouse
EAO was also inducible by the rat TGC but not the guinea pig TGC. Therefore,
the results suggest that the autoantigens that induce the mouse EAO are present in
not only the mouse TGC but also xenogenic TGC = the rat TGC. Analyses of the
common autoantigens responsible for EAO induction are now in progress.
P
Diethylhexyl phthalate
Di2ethylhexyl phthalate=DEHP
DEHP
A/J
8 0%0.01%0.1%DEHP 8
0.01%DEHP 0.1%DEHP
0.01%
0.1%DEHP
DEHP
MHC Class RTPCR
DEHP
IL10IFNγ
DEHP
P
The effect of cadmium on immunoenvironment in the testis
Yuki Ogawa 1,2 , Masahiro Itoh 1 , Shuichi Hirai 1 , Munekazu Naito 1 ,
Ning Qu 1 , Hayato Terayama 1 , Hidenobu Miyaso 2 , Yoshiharu Matsuno 2 ,
Masatoshi Komiyama 2 , Chisato Mori 2
1
Dept. Anat., Tokyo Med. Univ., 2 Dept. Bioenv. Med., Grad. Sch. Med., Chiba Univ.
Cadmium, one of various environmental toxicants, is known to suppress systemic
immunity and to injure the testicular capillary endothelia with resultant necrosis of
testicular tissues in mice and rats treated with high doses. Recently, it also became
evident that cadmium can affect the integrity of the bloodtestis barrier BTB,
the endocrine function of Leydig cells, apoptosis of germ cells and systemic
immunity, even on treatment with a low dose that does not induce spermatogenic
disturbance. Experimental autoimmune orchitis EAO, i.e., an organ
specific autoimmunity of the testis, can be induced by repeated immunization
with testicular antigens, and its pathology is characterized by lymphocytic
inflammation and spermatogenic disturbance. In the present study, we investigated
the morphological and functional changes of testes in mice treated with a low dose
of cadmium chloride CdCl 2 and also examined its toxicity as to susceptibility to
EAO.