Flow cytometric evaluation of the effects of doxorubicin on rat spermatogenesis

Suter, Laura; Bobadilla, María; Koch, Elke; Bechter, R.

doi:10.1016/s0890-6238(97)00019-1

Cited by 23 publications

(22 citation statements)

References 20 publications

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“…Recently, new approaches for detecting testicular toxicity with FCM have been reported; for example, detection of apoptosis (Krishnamurthy et al, 1998;Shiratsuchi et al, 1997), delay of spermatogenesis with BrdU staining and precise classification with the combination of other parameters (Suter et al, 1997b(Suter et al, , 1998a(Suter et al, and 1998b. Our procedure is suitable to combine with another staining kit, especially for a specific antibody, because the membrane of isolated cells remains intact due to no use of severe treatments with trypsin, detergent or pepsin.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, sensitivity is the major limitation of the FCM analysis in evaluating testicular toxicity. Several improved methods have been reported using in combination another parameter such as the somatic-specific antibody (Hittmair et al, 1994), the amount of mitochondria (Petit et al, 1995), or both of them (Suter et al, 1997a(Suter et al, , 1997b(Suter et al, , 1998a(Suter et al, , and 1998b.…”

Section: Introductionmentioning

confidence: 99%

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Assessment of quantitative dual-parameter flow cytometric analysis for the evaluation of testicular toxicity using cyclophosphamide- and ethinylestradiol-treated rats.

Katoh

Kitajima²,

Saga³

et al. 2002

J. Toxicol. Sci.

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-In the drug discovery process, effects to the human spermatogenesis must be fully evaluated before the first human trial. To estimate testicular toxicity, histopathological evaluation has been recommended in addition to the traditional mating procedure. However, it is laborious and time-consuming. Flow cytometric analysis (FCM) has also been applied to estimate testicular toxicity because of its speed, simplicity, and the objectivity of the data. Using cyclophosphamide (CP)-and ethinylestradiol (EE)-treated rat testis, we attempted to validate our dual-parameter, DNA ploidy and cell-size FCM, in a high-throughput toxicity study. Our results showed that CP damaged some spermatogonia and some early meiotic spermatocytes and EE caused severe decrease of spermatogenic cells except for spermatogonia as well as marked decrease of somatic cells, most probably Leydig cells. This is the first report discriminating between the changes of spermatogonia and that of somatic cells with FCM analysis. These results demonstrate that this method is a very useful and powerful tool to assess testicular toxicity, especially in high-throughput toxicological studies.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Assessment of quantitative dual-parameter flow cytometric analysis for the evaluation of testicular toxicity using cyclophosphamide- and ethinylestradiol-treated rats.

Katoh

Kitajima²,

Saga³

et al. 2002

J. Toxicol. Sci.

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“…Testicular damage by a toxicant is generally evaluated by analyzing parameters such as fertility, pregnancy outcome, testicular cell morphology, and sperm production and motility [7,9,12]. Traditional evaluation approaches also involve histopathologic examination of testicular tissue, which includes the description of several cell types, the determination of spermatogenic stages, and the detection of morphologic and cell-kinetic abnormalies in the spermatogenic process [13].…”

mentioning

confidence: 99%

“…Traditional evaluation approaches also involve histopathologic examination of testicular tissue, which includes the description of several cell types, the determination of spermatogenic stages, and the detection of morphologic and cell-kinetic abnormalies in the spermatogenic process [13]. However, these methods are subjective and time-consuming [10][11][12]. Moreover, the morphologic observation limits a local evaluation of the testicular tissue.…”

mentioning

confidence: 99%

“…Recently, flow cytometry (FCM) has become a useful tool for objective quantification of the types of testicular cell involved in spermatogenesis and it supplies valuable information for the detection of testicular toxicity [11,12]. As compared with current methods for the evaluation of spermatogenic impairment, FCM offers advantages in terms of objectivity, rapidity, analysis of large number of cells providing high statistical significance, and unbiased sampling of cells [10][11][12]. It also provides quantitative values for evaluating different cell types on the basis of their DNA ploidy/stainability level [10][11][12][13].…”

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confidence: 99%

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Flow Cytometric Assessment of Ethylene Glycol Monoethyl Ether on Spermatogenesis in Rats.

Yoon

Hong

Cho

et al. 2003

The Journal of Veterinary Medical Science

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ABSTRACT. The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells.

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