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Nuclear Morphometric Analysis of Leydig Cells of Male Pubertal Rats Exposed In Utero to Di(n-butyl) Phthalate.

Wakui S, Motohashi M, Satoh T, Shirai M, Mutou T, Takahashi H, Wempe MF, Endou H, Inomata T, Asari M - J Toxicol Pathol (2013)

Bottom Line: Nuclear pleomorphism of hyperplastic LCs is common and is considered to be continuous progressive degeneration.The results of the 5-wk-old DBP group were similar to those of the vehicle group; LC nuclei of the 7-wk-old DBP group showed normal ploidy and similar amounts of DNA.However, the size, elongation and peripheral chromatin aggregation parameters were significantly higher, and the reticular chromatin distribution and isolated chromatin aggregation parameters were significantly lower compared with the vehicle group.

View Article: PubMed Central - PubMed

Affiliation: Department of Toxicology, Laboratory Animal Science, and Veterinary Anatomy, Azabu University School of Veterinary Medicine, 1-17-71 Chuo, Sagamihara, Kanagawa 252-5201, Japan.

ABSTRACT
We recently reported that prenatal rat exposure to di(n-butyl) phthalate (DBP) induced Leydig cell (LC) hyperplasia after nine weeks (wks) of age, yet the number of LCs was similar to that of the vehicle group until seven weeks. Nuclear pleomorphism of hyperplastic LCs is common and is considered to be continuous progressive degeneration. Thus, computer-assisted image cell nuclear analysis of LCs was performed on 5- and 7-wk-old Sprague-Dawley (SD) rats whose dams had been administered DBP (i.g.) at 100 mg/kg/day or vehicle (corn oil) on gestation day 12 to 21. The results of the 5-wk-old DBP group were similar to those of the vehicle group; LC nuclei of the 7-wk-old DBP group showed normal ploidy and similar amounts of DNA. However, the size, elongation and peripheral chromatin aggregation parameters were significantly higher, and the reticular chromatin distribution and isolated chromatin aggregation parameters were significantly lower compared with the vehicle group. The present study quantitatively demonstrated nuclear morphological alterations in rat LCs at 7 wks old (puberty) due to the prenatal DBP administration before apparent LC hyperplasia developed.

No MeSH data available.


Related in: MedlinePlus

Representative features of Leydig cells nuclei of the 7-wk-old vehicle group (A) and 7-wk-old DBP group (B). DNA Feulgen stain without counterstain; bar = 25 μm.
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fig_001: Representative features of Leydig cells nuclei of the 7-wk-old vehicle group (A) and 7-wk-old DBP group (B). DNA Feulgen stain without counterstain; bar = 25 μm.

Mentions: Phthalates are chemicals used as plasticizers in polyvinyl chloride to impart flexibility and durability, and comprise up to 40% of the plastic volume. The phthalate esters, including di(n-butyl) phthalate (DBP), have an estrogenic or anti-androgenic effect on the development of the male reproductive system, and the specific primary cellular target of DBP has been considered to be testicular Leydig cells (LCs)22,23,24,25,26. Recently we reported that prenatal administration of DBP induced atypical Leydig cell (LC) hyperplasia at nine weeks and older, although the numbers and proliferative activities of LCs were similar to those of the vehicle group until seven weeks27,28. The progression from normal structure to hyperplasia has been considered a continuous event29. The degree of abnormal morphological aberration of individual cell nuclei is one of the important features in assigning a grade to pathological changes, and nuclear aberrations are always analyzed by a subjective assessment of chromatin pattern, size, and shape of the nuclei18,19,20,21,30,31. Progressive degeneration of LC nuclei before suffering hyperplasia is difficult to recognize by routine light microscopy, because the qualitative morphological alterations of LCs nuclei after prenatal DBP exposure are unclear (Fig. 1Fig. 1.


Nuclear Morphometric Analysis of Leydig Cells of Male Pubertal Rats Exposed In Utero to Di(n-butyl) Phthalate.

Wakui S, Motohashi M, Satoh T, Shirai M, Mutou T, Takahashi H, Wempe MF, Endou H, Inomata T, Asari M - J Toxicol Pathol (2013)

Representative features of Leydig cells nuclei of the 7-wk-old vehicle group (A) and 7-wk-old DBP group (B). DNA Feulgen stain without counterstain; bar = 25 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921929&req=5

fig_001: Representative features of Leydig cells nuclei of the 7-wk-old vehicle group (A) and 7-wk-old DBP group (B). DNA Feulgen stain without counterstain; bar = 25 μm.
Mentions: Phthalates are chemicals used as plasticizers in polyvinyl chloride to impart flexibility and durability, and comprise up to 40% of the plastic volume. The phthalate esters, including di(n-butyl) phthalate (DBP), have an estrogenic or anti-androgenic effect on the development of the male reproductive system, and the specific primary cellular target of DBP has been considered to be testicular Leydig cells (LCs)22,23,24,25,26. Recently we reported that prenatal administration of DBP induced atypical Leydig cell (LC) hyperplasia at nine weeks and older, although the numbers and proliferative activities of LCs were similar to those of the vehicle group until seven weeks27,28. The progression from normal structure to hyperplasia has been considered a continuous event29. The degree of abnormal morphological aberration of individual cell nuclei is one of the important features in assigning a grade to pathological changes, and nuclear aberrations are always analyzed by a subjective assessment of chromatin pattern, size, and shape of the nuclei18,19,20,21,30,31. Progressive degeneration of LC nuclei before suffering hyperplasia is difficult to recognize by routine light microscopy, because the qualitative morphological alterations of LCs nuclei after prenatal DBP exposure are unclear (Fig. 1Fig. 1.

Bottom Line: Nuclear pleomorphism of hyperplastic LCs is common and is considered to be continuous progressive degeneration.The results of the 5-wk-old DBP group were similar to those of the vehicle group; LC nuclei of the 7-wk-old DBP group showed normal ploidy and similar amounts of DNA.However, the size, elongation and peripheral chromatin aggregation parameters were significantly higher, and the reticular chromatin distribution and isolated chromatin aggregation parameters were significantly lower compared with the vehicle group.

View Article: PubMed Central - PubMed

Affiliation: Department of Toxicology, Laboratory Animal Science, and Veterinary Anatomy, Azabu University School of Veterinary Medicine, 1-17-71 Chuo, Sagamihara, Kanagawa 252-5201, Japan.

ABSTRACT
We recently reported that prenatal rat exposure to di(n-butyl) phthalate (DBP) induced Leydig cell (LC) hyperplasia after nine weeks (wks) of age, yet the number of LCs was similar to that of the vehicle group until seven weeks. Nuclear pleomorphism of hyperplastic LCs is common and is considered to be continuous progressive degeneration. Thus, computer-assisted image cell nuclear analysis of LCs was performed on 5- and 7-wk-old Sprague-Dawley (SD) rats whose dams had been administered DBP (i.g.) at 100 mg/kg/day or vehicle (corn oil) on gestation day 12 to 21. The results of the 5-wk-old DBP group were similar to those of the vehicle group; LC nuclei of the 7-wk-old DBP group showed normal ploidy and similar amounts of DNA. However, the size, elongation and peripheral chromatin aggregation parameters were significantly higher, and the reticular chromatin distribution and isolated chromatin aggregation parameters were significantly lower compared with the vehicle group. The present study quantitatively demonstrated nuclear morphological alterations in rat LCs at 7 wks old (puberty) due to the prenatal DBP administration before apparent LC hyperplasia developed.

No MeSH data available.


Related in: MedlinePlus