[Proteomic analysis of the testis and differential expression of Annexin A3 in hypospadiac rats].
OBJECTIVE
To investigate the effect of in utero exposure to di-n-butyl phthalate (DBP) on the protein expression in the penile tissue of hypospadiac rats, isolate and identify differentially expressed proteins, and determine the role of the differential expression of Annexin A3 in the development of hypospadia in the rat offspring after maternal exposure to DBP.


METHODS
Twenty pregnant SD rats were randomly assigned to an experimental group, intragastrically administered DBP at 800 mg/kg, and a control group, given soybean oil at 5 ml/kg, both for 5 days. Three days after birth, the penises of the newborn rats were removed, and the total protein extracted for 2D-electrophoretic separation and image analysis. Differentially expressed protein spots were screened and identified by mass spectrometry, and the changes in the expression of Annexin A3 detected by Western blotting and immunohistochemistry.


RESULTS
Thirty-one differentially expressed protein spots were screened, of which 17 were identified by mass spectrometry and the SwissProt database, including pyruvate kinase M2, alpha-enolase, and Annexin A3. Western blot showed that Annexin A3 was mainly located in the urethral epithelia and had a lower expression in the hypospadiac rats (1.851 +/- 0.014, n = 10) than in the controls (2.603 +/- 0.012, n = 10) (P < 0.05).


CONCLUSIONS
A pedigree of differentially expressed proteins in the penises of DBP-induced hypospadia and normal rats was established by the proteomic method. The differential expression of Annexin A3 may play an important role in the development of hypospadia.
Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China. doctorjkchang@163.com
Jun-Kai
JK
Chang
Wei
W
Zhang
Bai-Xin
BX
Shen
Yun-Fei
YF
Wei
Li-Feng
LF
Zhang
Yong
Y
Wang
Ning-Han
NH
Feng
D000818:Animals; D000831:Animals, Newborn; D017318:Annexin A3 / Q000378:metabolism*; D003993:Dibutyl Phthalate / Q000009:adverse effects; D004842:Epispadias; D005260:Female; D007021:Hypospadias / Q000378:metabolism*; D008297:Male; D018811:Maternal Exposure; D010413:Penis / Q000378:metabolism*; D011247:Pregnancy; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley
D020543:Proteome; D003993:Dibutyl Phthalate; D017318:Annexin A3