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Differential Expression of Genes: DISCUSSION(9)

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, the Ubel and Ube2d3 genes, also showed significant changes overall or in some segments. This indicates that the screening of the ubiquitin system provides valuable information about the effect of toxic exposure on the male reproductive system.

In the future, we will explore the simultaneous screening of all 19S, 20S, and 11S subunits as well as an expanded repertoire of ubiquitin-conjugating and deubiquitinating enzymes. Such an analysis could be greatly facilitated by automated transcriptional profiling—for example, by a microarray designed specifically to capture all gene products within the UPP.

In summary, our data indicate that DNB and THP exposures alter the expression of select genes within the UPP in a time-and dose-dependent manner. In particular, the Psmbl, Psmb2, Psmb5, and Ube2d3 genes that encode the constitutive proteasomal core and ubiquitin-activating enzyme UBE2 were affected. This change is most significant in the testis and corpus epididymis, but the magnitude of effect differs slightly by the administered toxicant. Such an altered gene expression may correlate with aberrant spermatogenesis in the testis and impair the processing of both normal and defective spermatozoa in the epididymis. Transcriptional profiling and flow cytometric analysis of the UPP thus capture the subtle effects of reproductive toxicity not observed by conventional histology and functional cytology and offer a prospective new tool to detect and manage reproductive toxicology. In addition to mapping the effect of reprotoxic exposure on the expression of a specific subset of functionally related genes in the testis and epididymis, the present study showed a gradient in the expression of the UPP pathway genes that, for most genes, seemed to follow the pattern of testis > caput > corpus > cauda or caput > testis > corpus > cauda.

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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