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Differential Expression of Genes: RESULTS(12)

UBE2 showed a distinct perinuclear focus in the principal cells of the control epididymis, reminiscent of endoplasmic reticulum localization pattern (Fig. 9B). Some of the principal cells showed a nuclear localization of the ubiquitinated protein species recognized by antibody MK12-3 (Fig. 9B). The endoplasmic reticulum-like localization pattern was absent from the DNB epididymis (Fig. 9B’), while the nuclear accumulation of anti-ubiquitin immunoreactive proteins was more pronounced (Fig. 9B0, insert). UCHL1 showed similar patterns in both the control (Fig. 9C) and DNP-exposed epididymis (Fig. 9C0).

In both types of tissue, a diffuse labeling was present in the cytosol of the principal cells, while a concentrated labeling that coincided with that of the ubiquitinated proteins was seen in the clear cells. A similar distribution pattern was followed by the proteasomes in the principal and clear cells of the control (PSMA/PMSB; Fig. 9D) and DNB-exposed (Fig. 9D0) epididymis. In this example, proteasomes were identified by a polyclonal antibody that recognized multiple 20S proteasomal a and p subunits. Noteworthy was the accumulation of ubiquitin and/or ubiquitinated substrates on the apical surface of the epididymal epithelium, recognized by anti-ubiquitin antibody KM691. Negative control slides did not show any of the patterns described above for the testis or epididymis in the control or DNB-exposed animals.

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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