2002 Fiscal Year Final Research Report Summary
Production of nitric oxide by Sertoli cell and its influence upon the cells
| Project/Area Number |
12671572
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | National Institute of Radiological Sciences |
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Principal Investigator |
ONODA Makoto Institute of Radiological Sciences, Redox Regulation Research Group, Senior Researcher, レドックス制御研究グループ, 主任研究員 (20260234)
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| Project Period (FY) |
2000 – 2002
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| Keywords | nitric oxide / nitric oxide synthase / Sertoli cell / spermatogenesiss / tight junction / blood-brain barrier / occludin / Z0-1 |
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| Research Abstract |
Rat seminiferous epithelial cells (Sertoli cells : SC) isolated from immature rats were cultured either in the bicameral chamber system or on 15-mm round glass cover slide to validate the effect of nitric oxide (NO) on the protein secretion from the SC and the function of permeability barrier (tight junction) formed by the SC monolayer. The results were as follows.
1. The amount of NO produced spontaneously by SC in culture was relatively minute.
2. The NO concentration in the conditioned media was not significantly increased by the addition of LPS (1 μg/ml) to the basal reservoir of the culture system.
3. The electrical resistance (ER) of SC monolayer was not influenced by the addition of the exogenous NO that released from a NO donor (NOC18).
4. The amount of the total transferrin (TF) secreted by SC was not significantly different statistically among the control and the NOC18 treated epithelial sheets. However, the polarity of the TF secretion by SC was significantly lower for the NO-do
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nor contained cultures. The secretion ratio (apical/basal=1.86 ± 0.22) of TF during 2-day collection period was significantly declined to 1.26 ± 0.04 and 1.47 ± 0.08 in the culture with the exogenously added NOC18 (50 and 100 μM), respectively.
5. Immunostaining for occludin and ZO-1 in untreated-SC showed continuous labeling around the cell periphery in the region of the cell-cell junctional complex, and these colocalized with TJ-associated F-actin filaments.
6. However, incubation with NOC 18 (200 μM and 400 μM) caused complete disorganization of occludin staining and partial disruption of ZO-1 staining within 48 h.
7. Furthermore, descending expressions of occludin, ZO-1 and cortactin were observed in a dose-dependent manner of NOC 18 in NO donor treated SC.
These results suggest that NO produced excessively within testis during acute or chronic pathophysiological conditions disrupts TJ-associated proteins of SC and that NO may perturb blood-testis barrier formation and in turn the regulation of normal spermatogenesis. Less
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Research Products
(12 results)
