scholarly journals Time-Dependent and Compartment-Specific Effects of In Utero Exposure to Di(n-butyl) Phthalate on Gene/Protein Expression in the Fetal Rat Testis as Revealed by Transcription Profiling and Laser Capture Microdissection

2007 ◽  
Vol 97 (2) ◽  
pp. 520-532 ◽  
Author(s):  
S. Plummer ◽  
R. M. Sharpe ◽  
N. Hallmark ◽  
I. K. Mahood ◽  
C. Elcombe
Keyword(s):  
Protein Expression ◽  
Laser Capture Microdissection ◽  
Time Dependent ◽  
In Utero Exposure ◽  
Transcription Profiling ◽  
Rat Testis ◽  
Specific Effects ◽  
Fetal Rat ◽  
Butyl Phthalate ◽  
Laser Capture

In utero exposure of high-dose di-n-butyl phthalate resulted in opposite effects on testicular cell apoptosis in late embryonic and pubertal male rat offspring

2017 ◽  
Vol 36 (12) ◽  
pp. 1236-1247 ◽  
Author(s):  
H Shen ◽  
K Liao ◽  
H-F Wu ◽  
H-C Lu ◽  
Y Li ◽  
...  
Keyword(s):  
Western Blot ◽  
Cell Apoptosis ◽  
In Utero ◽  
Male Rat ◽  
In Utero Exposure ◽  
High Dose ◽  
Testicular Cell ◽  
Rat Testis ◽  
Rat Offspring ◽  
Butyl Phthalate

Objective: To investigate the effects of in utero exposure to high-dose di- n-butyl phthalate (DBP) on testicular cell apoptosis in late embryonic and pubertal male rat offspring. Methods: Twenty pregnant Sprague-Dawley (SD) rats were divided into two groups. During gestation day (GD) 12 to GD 19, control group was given 1 ml day−1 of olive oil and experimental group was given DBP 500 mg kg−1 day−1 by gavage. On GD 19.5 and postnatal day (PND) 45, the testes were removed. Morphological analysis of the testes was observed by transmission electron microscopy and hematoxylin and eosin (H&E) staining. Testicular cell apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The expression of Bcl-2, Bax, and p53 was presented by immunohistochemistry (IHC) and western blot. Data of the two groups was compared using independent samples t-test and Mann–Whitney test by SPSS 20.0. Results: H&E staining showed that spermatogenetic cells were significantly decreased in DBP exposed pubertal rat testis. The apoptosis index of testes in DBP-treated group was significantly lower on GD 19.5 but higher on PND 45 than that of the controls ( p < 0.01). IHC and western blot revealed significantly increased expression of Bcl-2 in GD 19.5 rat testis and Bax and p53 in PND 45 rat testis after DBP exposure, compared with the control ( p < 0.05). Conclusion: In utero exposure of high-dose DBP resulted in opposite effects on testicular cell apoptosis in late embryonic and pubertal rat offspring. The overexpression of Bcl-2, Bax, and p53 might be related to the occurrence of abnormal apoptosis and finally produce male infertility.

Differential expression of peroxiredoxin 6 in fetal rat testis following in utero exposure to di(nbutyl) phthalate

Toxicology ◽  
2007 ◽  
Vol 240 (1-2) ◽  
pp. 86-95 ◽  
Author(s):  
Wei Zhang ◽  
Hua Shen ◽  
Long Ma ◽  
Baixin Shen ◽  
Zicheng Xu ◽  
...  
Keyword(s):  
Differential Expression ◽  
In Utero ◽  
In Utero Exposure ◽  
Peroxiredoxin 6 ◽  
Rat Testis ◽  
Fetal Rat

Novel method for predicting chemoresistance to paclitaxel in epithelial ovarian cancer patients

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 15007-15007
Author(s):  
D. Silasi ◽  
T. J. Rutherford ◽  
P. E. Schwartz ◽  
R. Chen ◽  
A. Alvero ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Protein Expression ◽  
Western Blot ◽  
Epithelial Ovarian Cancer ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Laser Capture Microdissection ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Laser Capture

15007 Background: No available test exists to guide the selection of effective chemotherapeutic regimen in recurrent ovarian cancer. Preliminary studies in our lab have identified a protein, MyD88, a major component in the inflammatory pathway, to be highly expressed in epithelial ovarian cancer cells that exhibit primary or acquired Paclitaxel chemoresistance. The objective of this study was to develop a sensitive approach that can detect expression of MyD88 in ovarian cancer tissue. We report the development of a test based on Laser capture microdissection that allows detection of MyD88 in a 6000 cell sample. Methods: Tumor tissue was obtained at surgery from epithelial ovarian cancer patients and snap-frozen in liquid nitrogen. Eight micron sections were prepared on polyethylene covered glass slides and tumor cells were dissected with a Laser capture microdissection system. Protein expression was detected by Western blot analysis. Results: Protein expression was detected by Western blot analysis in 1000 microdissected cells. An inverse correlation was observed between MyD88 expression in tumor cells and clinical response to Paclitaxel. Furthermore, this method allows the isolation of CD-45 positive cells from the tumor and analysis of their protein expression. Conclusions: We describe for the first time a method that will allow us to predict chemoresistance. Laser capture microdissection is a powerful technique that can be used to study the protein profile of each of the cellular components present in the tumor microenvironment. This technique will facilitate our understanding of the proteins necessary for tumor growth and may help to identify novel markers or potential protein targets. No significant financial relationships to disclose.

scholarly journals Reconstitution of rat fetal testis during the masculinisation programming window induces focal dysgenesis consistent with testicular dysgenesis syndrome

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ellie Smart ◽  
Joni Macdonald ◽  
Lee B. Smith ◽  
Rod T. Mitchell
Keyword(s):  
Testicular Dysgenesis Syndrome ◽  
Fetal Testis ◽  
Specific Effects ◽  
Fetal Rat ◽  
Testicular Dysgenesis ◽  
Seminal Vesicle Weight ◽  
Consistent Feature ◽  
Butyl Phthalate ◽  
Testis Tissue

Abstract Focal dysgenesis is a consistent feature of testicular dysgenesis syndrome (TDS) in humans. Rodent studies show that perturbation of androgens (e.g. following phthalate exposure) during a fetal masculinisation programming window (MPW) predisposes to a TDS phenotype. This study aimed to determine whether dissociation and reconstitution of rat fetal testis tissue during the MPW can be used to model and manipulate seminiferous cord development, including induction of focal dysgenesis, as described in TDS. Dissociated fetal rat testes were xenotransplanted subcutaneously into recipient mice for 4 weeks. Transplanted mice were treated with vehicle or di-n-butyl-phthalate (DBP, a plasticising chemical known to induce testicular dysgenesis in vivo in rats). Testosterone production by the transplants was measured in recipient mice and immunofluorescence was performed on the retrieved transplants to identify features consistent with focal testicular dysgenesis. Re-aggregation of rat fetal testis tissue xenotransplants during the MPW results in reconstitution of seminiferous cords. Features of focal testicular dysgenesis were present in re-aggregated testis, including ectopic Sertoli cells and intratubular Leydig cells (ITLCs). DBP exposure of recipient mice reduced androgen-dependent seminal vesicle weight (8.3 vs 26.7 mg; p < 0.05), but did not enhance features of focal dysgenesis including number of ITLCs (0.07 vs 0.10 cells/mm2; p > 0.05). We conclude that seminiferous cord reformation during the MPW results in development of focal dysgenesis. The system may be used to separate specific effects (e.g. androgen suppression) of individual chemical exposures from other mechanisms that may be conserved in TDS.

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