RESULTS(3)

GGT catalytic activity (Fig. 7) and GGT protein level (Fig. 8) in the initial segment decreased after 3-day EDL but returned to the sham control value after incubations with TPA but not with FCS.

Presence of bFGF-Like Proteins in RTF and the Rat Epididymis

Using a bFGF pAb, 16-kDa and 43-kDa protein bands were detected in rRTF (20 |jLg). However, no protein bands were detected in oRTF or no-ABP oRTF preparations using this bFGF antibody (Fig. 9A). Using a bFGF mAb, 16-, 24-, 43-, and 57-kDa proteins were detected in rRTF, and 34-kDa and 67-kDa proteins were detected in no-ABP oRTF and oRTF by Western blot analysis (Fig. 9B).

Using the bFGF pAb, the 43-kDa bFGF-like protein was present in luminal fluids (5 |xg total proteins) from rat rete testis, initial segment, caput, and cauda (Fig. 10A). The level of this 43-kDa bFGF-like protein remained constant along the epididymal duct (Fig. 10A). A 16-kDa protein band was also detected in 1-2 (jlI (25-50 jxg total proteins) of luminal fluids from the initial segment, caput, and cauda (data not shown). flovent inhaler

As shown in Figure 10B, the 43-kDa bFGF-like protein was also present in the homogenates from the initial segment, caput, corpus, and cauda. The level of this protein increased from the proximal to the distal epididymal regions (Fig. 10B). In the initial segment following 3-day EDL, the level of this bFGF-like protein decreased in comparison to the level in the sham control (Fig. 10C). The 43-kDa bFGF-like protein level in the initial segment did not return to the sham control value after treatment with rRTF (Fig. 10D).
Fig7Regulation of gamma-glutamyl
FIG. 7. The recovery of decreased GGT catalytic activity in EDL 3 D initial segments by TPA. Initial segments from EDL 3 D-operated rats were incubated with or without TPA or 10% FCS at 35°C for 6 h. The sham-treated initial segment was used as a control. GGT catalytic activity was determined in the homogenates of the treated tissues and expressed as a percentage of GGT catalytic activity in the sham-treated initial segment in each experiment. Data shown represent mean percentage ± SEM from three individual experiments in each group. Mean percentages for GGT catalytic activity with different letters were significantly different (p < 0.05).

Fig7Regulation of gamma-glutamyl
FIG. 8. The recovery of the decreased GGT protein level in EDL 3 D initial segments by TPA. Initial segments from EDL 3 D-operated rats were incubated with or without different concentrations of TPA or 10% FCS at 35°C for 6 h. The sham-treated initial segment was used as a control. The homogenates of treated initial segments were subjected to Western blot analyses. Fifty micrograms of homogenate were separated by 12% SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was then probed with rabbit anti-GGT antibody (1:500) for 3 h at room temperature. Molecular masses, kDa, of detected GGT proteins are indicated on the left.

Fig9Regulation of gamma-glutamyl
FIG. 9. The presence ot bFGF-like proteins in rKl F as revealed by Western blot analyses.

Fig10Regulation of gamma-glutamyl
FIG. 10. The presence of bFGF-like proteins in the rat epididymis and the effect of EDL on the bFGF-like protein level in the initial segment as revealed by Western blot analyses.