Expression of TASK-1 channel in mouse Leydig cells

다국어 초록 (Multilingual Abstract)

Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K+ channel-related Acid-Sensitive K+-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker’s effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK- 1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LHinduced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

목차 (Table of Contents)

• INTRODUCTION
• MATERIALS AND METHODS
• Chemicals
• Animals and testis isolation
• Hematoxylin and eosin (H&E) staining
• Immunohistochemistry (IHC)
• Cell culture
• Total RNA extraction
• Reverse transcriptase-polymerase chain reaction (RTPCR)
• Western blot analysis
• Live/dead cell staining
• Cell viability assay
• Measurement of testosterone concentration
• Statistical analysis
• RESULTS
• TASK-1 is expressed in Leydig cells and Sertoli cellswithin the testes of mice
• TASK-1 is more highly expressed in TM3 cellscompared to TM4 cells
• Blocking TASK-1 induces death in TM3 cells
• DISCUSSION
• REFERENCES