24 February 2009 Verifying of endocrine disruptor chemical affect to the mouse testes: can raman spectroscopy support histology study?
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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.
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Bibin Bintang Andriana, Bibin Bintang Andriana, Yusuke Oshima, Yusuke Oshima, Sota Takanezawa, Sota Takanezawa, Tat Wei Tay, Tat Wei Tay, Catherine Linda Rosawati Soeratman, Catherine Linda Rosawati Soeratman, Mohammad Shah Alam, Mohammad Shah Alam, Hiroki Mitsuoka, Hiroki Mitsuoka, Xiao Bo Zhu, Xiao Bo Zhu, Toshiaki Suzuki, Toshiaki Suzuki, Yuko S. Yamamoto, Yuko S. Yamamoto, Naoki Tsunekawa, Naoki Tsunekawa, Yoshiakira Kanai, Yoshiakira Kanai, Masamichi Kurohmaru, Masamichi Kurohmaru, Hidetoshi Sato, Hidetoshi Sato, } "Verifying of endocrine disruptor chemical affect to the mouse testes: can raman spectroscopy support histology study?", Proc. SPIE 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems, 71670Y (24 February 2009); doi: 10.1117/12.808700; https://doi.org/10.1117/12.808700
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