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Proceedings Paper

Verifying of endocrine disruptor chemical affect to the mouse testes: can raman spectroscopy support histology study?
Author(s): Bibin Bintang Andriana; Yusuke Oshima; Sota Takanezawa; Tat Wei Tay; Catherine Linda Rosawati Soeratman; Mohammad Shah Alam; Hiroki Mitsuoka; Xiao Bo Zhu; Toshiaki Suzuki; Yuko S. Yamamoto; Naoki Tsunekawa; Yoshiakira Kanai; Masamichi Kurohmaru; Hidetoshi Sato
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Paper Abstract

One of suspect environmental endocrine disruptors that affect mouse male reproduction by altering the morphology of Sertoli cells and spermatogenic cells is phthalate. The effects of mono(2-ethylhexyl)phthalate (MEHP), one of metabolites of di(2-ethylhexyl)phthalate , on immature mouse testes in vivo were examined. We have recently shown that MEHP induced Sertoli cells necrosis and spermatogenic cells apoptosis in mice by TUNEL method, F-actin staining, and ultrastructural study, but there is no data for biochemical changing of testes due to those methods could not explore. To verify in detail of it, we conducted Raman spectroscopy study with 785 nm wavelength laser line, 50mW of laser power and 3 minutes of exposure time to analysis the MEHP-treated testicular tissue, which has been fixatived by 4% paraformaldehyde (PFA). Five weeks old (5 w.o) male mice were used in this experiment. As the results, the alterations were observed by Raman spectroscopy that there are significantly differences of DNA, actin filament, type IV collagen and amide I between control group (0 μM MEHP) and treatment group (100 μM MEHP). These results significantly support histology staining observation (such as the apoptotic spermatogenic cells which is associated with DNA fragmentation and F-actin disruption) and ultrastructural observation (such as mitochondria rupture and disintegration of nucleus membrane). Raman spectroscopy can be used for 4% PFA-fixatived tissue observation. However, we recommend that Raman spectroscopy may be able to be expanded as an armamentarium not just for the clarification of histology staining and ultrastructural study, but furthermore, it may be as a non-invasion assessment for screening animal tissue toxicity of chemical in future.

Paper Details

Date Published: 24 February 2009
PDF: 11 pages
Proc. SPIE 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems, 71670Y (24 February 2009); doi: 10.1117/12.808700
Show Author Affiliations
Bibin Bintang Andriana, The Institute of Physical and Chemical Research (RIKEN) (Japan)
The Univ. of Tokyo (Japan)
Yusuke Oshima, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Sota Takanezawa, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Tat Wei Tay, The Univ. of Tokyo (Japan)
Catherine Linda Rosawati Soeratman, The Univ. of Tokyo (Japan)
Mohammad Shah Alam, The Univ. of Tokyo (Japan)
Hiroki Mitsuoka, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Xiao Bo Zhu, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Toshiaki Suzuki, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Yuko S. Yamamoto, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Naoki Tsunekawa, The Institute of Physical and Chemical Research (RIKEN) (Japan)
Yoshiakira Kanai, The Univ. of Tokyo (Japan)
Masamichi Kurohmaru, The Univ. of Tokyo (Japan)
Hidetoshi Sato, The Institute of Physical and Chemical Research (RIKEN) (Japan)


Published in SPIE Proceedings Vol. 7167:
Frontiers in Pathogen Detection: From Nanosensors to Systems
Philippe M. Fauchet, Editor(s)

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