Validation of Flow Cytometry Analysis in the Objective Assessment of Spermatogenesis: Comparison to the Quantitative Testicular Biopsy

Hirsch, Irvin H.; McCue, Peter; Kulp-Hugues, Deborah; Sedor, John; Flanigan, Maureen

doi:10.1016/s0022-5347(17)35480-0

Cited by 23 publications

(9 citation statements)

References 28 publications

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“…The flow cytometric analysis of quantitative changes in cellular composition provides an exact estimation of mitotic and meiotic steps in the testicular tissue. DNA flow cytometry has been established as a sensitive and precise method to determine parameters of spermatogenesis (Hacker-Klom et al 1986, Hellstrom et al 1990, Suresh et al 1992, Hirsch et al 1993, Spano and Evenson 1993. The discrimination between somatic and spermatogenic cells by dual staining of testicular cells with propidium iodide and FITC-labelled anti-vimentin antibody provides additional information about the proportions of germinative and somatic cells , Blottner and Roelants 1998.…”

Section: Discussionmentioning

confidence: 99%

Studies on the European hare. 56. Seasonal variation of testicular activity in European brown hare Lepus europaeus

Blöttner¹,

Faber²,

Roelants³

2000

Acta Theriol.

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Testicular activity in brown hare Lepus europaeus Pallas, 1778 was studied during the annual cycle. Testicular mass, spermatozoa and testosterone were estimated. Percentages of haploid, diploid and tetraploid cells were monitored using DNA flow cytometry and the proportions of somatic and spermatogenetic cells were determined after selective labelling of somatic cells. Testis mass was high from January to July and declined thereafter to the nadir in September. Testis growth was reactivated significantly in December. Changes in testis mass corresponded with the spermatogenic efficacy (spermatozoa/g testis). High spermatogenic activity was characterized by intensive meiotic transformation of spermatocytes to spermatids, high percentages of haploid cells and low proportions of cells in the G2/M phase of mitosis. Proportions of haploid cells declined rapidly during the testis involution. Spermatogenesis was newly activated from November. The proportions as well as the ratios of spermatogenic and somatic cells differed significantly between the periods of testis involution and recrudescence. Testosterone level showed a pronounced increase in autumn preceding the intensification of spermatogenesis; the lowest concentration was found during prominent testis involution in August. The results suggest that the regulation of seasonal testicular activity is characterized by co-ordinated shifts in the relationships between mitosis, meiosis and testosterone production.

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Section: Discussionmentioning

confidence: 99%

Studies on the European hare. 56. Seasonal variation of testicular activity in European brown hare Lepus europaeus

Blöttner¹,

Faber²,

Roelants³

2000

Acta Theriol.

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“…In rodents, testicular toxicity has been evaluated with FCM using many kinds of chemicals and compounds such as nitrobenzene (Iida et al, 1997), vin-blastine (Jagetia et al, 1996), methoxyacetic acid (Spanò et al, 1991;Krishnamurthy et al, 1998;Suter et al, 1998b) and 2-bromopropane (Son et al, 1999). In clinical studies, FCM analysis has also been applied for the diagnosis of human male infertility and has been proven to correlate highly with the result of histopathology (Hellstrom et al, 1990;Lee and Choo, 1991;Hittmair et al, 1992;Hirsch et al, 1993;Giwercman et al, 1994;Kostakopoulos et al, 1997).…”

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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-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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“…I. M. El-Ashmawy concluded that grape seed extracted is a useful herbal remedy, especially for controlling oxidative damages and is considered as a potent protective agent against hepatotoxicity 28 . Several lines of evidence demonstrated that, grapes seed proanthocyanidins exhibited in vivo hepatoprotective and anti-fibrogenic effects against liver injury and act as free radical's scavengers and protective liver damage 20 .…”

Section: Discussionmentioning

confidence: 99%

“… Group 3: GSE rats: Rats were administrated orally GSE (100 mg/kg b·w/day) diluted with saline solution; for three weeks 28,45 .…”

Section: Preparation Of Grape Seed Extracts (Gse)mentioning

confidence: 99%

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2016

IJPSR

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Aim: The present study has been undertaken to investigate the therapeutic effect of GSE (Grape seed extracts) against Tamoxifen (TAM), induced hepatotoxicity in rat. Methods: The rats were divided into four groups • Group 1: rats were injected intraperitoneally (i.p.) with saline for seven days.• Group 2: Rats were treated with TAM in a dose of 45 mg/kg b·w/day, i.p., for seven successive days.• Group 3: Rats were administrated orally GSE (100 mg/kg b·w/day) for three weeks. Group 4: rats were injected (i.p.) (45 mg/kg b·w/day) of Tamoxifen for seven days, then treated daily with a single dose of GSE (100 mg/kg b·w/day) for three weeks respectively. Results: GSE reduced necrosis in the TAM-treated rat. And significantly increased (p<0.05) the levels of MDA and PCC, while the level of GSH was significantly (P<0.05) decreased. Treatment with GSE significantly (P<0.05) reduced. Tamoxifen significantly decreased (P<0.05) the level of NO. GSE significantly increases (P<0.05, Table 2) NO level. Flowcytometric analysis in liver cells, significant increases in apoptotic cells treated by TAM and decreases in the group exposed to TAM and treated with GSE. Conclusion: This study suggests that GSE possesses anti-oxidant effects against TAM toxicity.

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Validation of Flow Cytometry Analysis in the Objective Assessment of Spermatogenesis: Comparison to the Quantitative Testicular Biopsy

Cited by 23 publications

References 28 publications

Studies on the European hare. 56. Seasonal variation of testicular activity in European brown hare Lepus europaeus

Studies on the European hare. 56. Seasonal variation of testicular activity in European brown hare Lepus europaeus

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

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