Archive for the ‘Epididymis by Testicular’ Category


Regulation of Gamma-Glutamyl Transpeptidase: RESULTS(3)

Nov 27, 2012 Author: Walter Mcneil | Filed under: Epididymis by Testicular

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

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  • Regulation of Gamma-Glutamyl Transpeptidase: RESULTS(2)

    Nov 26, 2012 Author: Walter Mcneil | Filed under: Epididymis by Testicular

    When the initial segment after 3-day EDL was treated with 40% > 10-kDa oRTF, GGT catalytic activity returned to the sham control value (Fig. 3B). However, treatment with < 10-kDa oRTF did not recover the decreased GGT catalytic activity (Fig. 3B). GGT catalytic activity in the initial segment after 3-day EDL returned to sham control values after treatment with 20% rRTF (Fig. 3C).

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  • Regulation of Gamma-Glutamyl Transpeptidase: RESULTS(1)

    Nov 25, 2012 Author: Walter Mcneil | Filed under: Epididymis by Testicular

    RESULTS(1)

    Effects of EDL on GGT Catalytic Activity in Various Regions of the Rat Epididymis

    GGT catalytic activity was measured in epididymal tissue homogenate from rats that had previously undergone unilateral EDL and a sham operation. For sham-operated rats, the levels of GGT catalytic activity were initial segment > caput > corpus = cauda > testes (Fig. 1). GGT catalytic activity in the initial segment decreased by 3 days and 5 days of EDL, but did not differ significantly from that in controls after 1 day and 14 days of EDL. In the caput and corpus regions, GGT catalytic activity did not decrease following 1 day, 3 days, and 5 days of EDL, but decreased by 14 days of EDL. GGT catalytic activity in cauda and testis remained unchanged after EDL for the experimental time periods. buy asthma inhalers (more…)

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  • For immunoblotting with bFGF antibodies, the filters were blocked with 1% BSA and 1% nonfat dry milk in TBST and then incubated with 1:500 bFGF monoclonal antibody (mAb; Sigma) or with 1:400 bFGF pAb (Chemi-con International, Temecula, CA). After being washed with TBST twice, the filters were incubated with horseradish peroxidase-conjugated goat anti-mouse antibody (Santa Cruz Biotech) or horseradish peroxidase-conjugated goat anti-rabbit antibody (Santa Cruz Biotech). After extensive washing with TBST, the filters were developed by the Boehringer-Mannheim (Indianapolis, IN) chemilumines-cence method described in the manufacturer’s protocol. The Western blotting study of bFGF-like protein in the luminal fluid from various epididymal regions was repeated twice, and the other experiments were repeated on samples from at least three individual experiments. buy flovent inhaler

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  • MATERIALS AND METHODS(3)

    RIA

    The concentrations of testosterone in oRTF preparations were determined using an ImmuChem testosterone 125I RIA kit (ICN Biomedicals, Costa Mesa, CA) according to the manufacturer’s protocol. asthma inhalers

    GGT Assay

    GGT catalytic activity assay was performed as described previously . Protein concentrations of samples were determined by the Bio-Rad (Richmond, CA) protein assay. For the comparative study of GGT catalytic activity among various tissues and of the effects of EDL on the GGT catalytic activity in these tissues, GGT catalytic activity was expressed as nmol/min per milligram protein. For the observation of whether RTF preparations and bFGF could recover GGT catalytic activity, the GGT catalytic activity was expressed as percentage of GGT catalytic activity in relation to that of each sham control.

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  • Preparations of Luminal Fluids from the Rat Initial Segment, Caput, and Cauda

    Luminal fluid from the initial segment, caput, and cauda of adult rats was collected as described previously . This luminal fluid was centrifuged at 3000 rpm in an Eppendorf microcentrifuge to remove the spermatozoa. The supernatant was collected and stored at — 80°C before use. buy antibiotics online

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  • MATERIALS AND METHODS(1)

    Animals

    Normal adult male retired breeder Sprague-Dawley rats (450-550 g; Hilltop Laboratories, Philadelphia, PA) were maintained on a 12L:12D cycle, with free access to food and water, in the University of Virginia vivarium. All experiments in this study were approved by the University of Virginia Animal Welfare Committee.

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  • Previous studies have shown that macromolecules in ovine RTF (oRTF) are beneficial for maintenance of the ability to metabolize testosterone by cultured principal cells from the proximal epididymal regions , and these macromolecules can stimulate protein secretion in the initial segment of the rat epididymis . Ovine RTF contains a growth factor , and a number of growth factors are produced by the testis . Therefore, we hypothesize that testicular factors such as growth factors are involved in the regulation of GGT catalytic activity and GGT protein level in the initial segment.

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