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Differential Expression of Genes: DISCUSSION(1)

DISCUSSION(1)

The present study demonstrates the reproductive toxic and spermatotoxic effects of THP and DNB by conventional means (histology and DIC microcopy) and by novel approaches, including flow cytometric analysis of isolated epididymal spermatozoa and transcriptional profiling of select gene products within the proteolytic UPP. Our goal was to determine if transcriptional analysis and flow cytometry could capture minor changes in the male reproductive system of rats exposed to reprotoxic chemicals. We were particularly interested in examining threshold toxic exposures at which a conventional histological examination did not reveal any pathology.

No damage to testicular or epididymal tissues was identified by histology prior to Day 30 of THP exposure (Supplemental Table 1). Similarly, only some of the 2-mg/kg DNB-exposed animals showed testicular or epididymal histopathology (Supplemental Table 2).

Contrary to ambiguous histopathological findings, the THP-induced sperm damage and breakdown were manifested by altered flow cytometric median values of ubiquitin on Days 30 and 42 (Fig. 3). While the difference on Day 18 was not significant (P = 0.79), several treated samples already exhibited a flattened, shifted flow cytometry histogram, suggestive of sperm fragmentation, and a subpopulation of highly fluorescent cells, suggestive of increased sperm ubiquitination (see Fig. 3B).

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Differential Expression of Genes: DISCUSSION(9)

While we found that both constitutive p subunits and their inducible counterparts were expressed in the testis and epididymis, only the constitutional subunits showed significant expression changes in the tissues of the exposed animals. Products of two of the three examined UPP enzyme genes not directly associated with the proteasome, … Continue reading

Differential Expression of Genes: DISCUSSION(8)

However, several other genes showed significant expression level changes in certain segments alone after either THP or DNB exposure. This indicates that THP and DNB cause significant changes in gene expression in specific compartments of the male reproductive system. For example, Psmb8 showed a P-value of 0.156 between the control … Continue reading

Differential Expression of Genes: DISCUSSION(7)

Inducible proteasomal core subunits PSMB9, PMSB8, and PSMB10 replace their constitutive counterparts PMSB5, PMSB1, and PMSB2 in the professional antigen-presenting cells but are also present in other cell types, such as eye lens cells and the sperm acrosome. The expression of inducible subunit Pmsb8 as measured by semiquantitative RT was … Continue reading

Differential Expression of Genes: DISCUSSION(6)

We have shown that defective spermatozoa become ubiquitinated in the caput epididymis, presumably by the enzymatic ubiquitination machinery residing within epididymal fluid. The percentage of defective spermatozoa is reduced after passage through the corpus epididymis. Since proteasomes have also been detected in the epididymal fluid, it is possible that defective … Continue reading

Differential Expression of Genes: DISCUSSION(5)

Alzheimer disease has been linked to the aberrant transcription of the ubiquitin-B (UB-B) gene, caused by a +1 frame-shift during transcription. This misreading results in the translation of the dysfunctional UB-B +1 protein with the elongated C-terminus not capable of ligation to a substrate protein. Consequently, the amyloid protein within … Continue reading