ISSN:
1573-4919
Keywords:
testis
;
signalling
;
inositol lipid
;
endothelin
;
purinergic receptors
;
Sertoli cells
;
peritubular myoid cells
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
Notes:
Abstract The testis is a complex organ in which local control is achieved by signalling between its constituent cells. Herein we describe the responses of cultured rat testicular cells and a mouse Sertoli cell-line to stimulation by endothelin and ATP, and elsewhere we have shown that rat peritubular myoid cells possess phosphoinositidase C-coupled Vla-vasopressin receptors identical to those of liver (Howl, J.et al, 1995, Endocrinology 136: 2206–2213). 1. Peritubular myoid cells from pre-pubertal rats responded through ETA receptors with PtdIns(4,5)P 2 hydrolysis [EC50 for endothelin-1 (ET-1)∼0.4 nM], elevation of intracellular [Ca2+], and tyrosine phosphorylation of a variety of cellular proteins. They also showed enhanced adenylate cyclase activity, with an EC50 for ET-1 of∼3 nM, also through ETA receptors. Pharmacological elevation of [cAMP] did not immediately change the ET-1-stimulated formation of inositol phosphates, but attenuated the response after several hours. 2. Pre-pubertal rat Sertoli cells showed no detectable responses to ET-1, but responded to FSH with elevated [cAMP] and to ATP with PtdIns(4,5)P 2 hydrolysis. PtdIns(4,5)P 2 hydrolysis was equally responsive to ATP and UTP, and so appears to be activated by P2U-purinergic receptors. This response was enhanced by protein kinase C inhibition and attenuated by PKC activation. 3. Despite its lack of effect on rat Sertoli cells in primary culture, ET-1 provoked PtdIns(4,5)P 2 hydrolysis in the TM4 murine Sertoli cell line (EC50∼0.6 nM), and this response was negatively regulated by protein kinase C activation. 5. No receptorstimulated activation of phosphoinositase C was detected in ‘germ cell’ populations, but the non-specific G protein activator AIF 4 − provoked inositol phosphate accumulation in these cells, so demonstrating their potential to respond through yet to be identified G protein-coupled receptors with phosphoinositidase C activation. 6. Immunoblotting studies showed the presence in rat testis of phosphoinositidase C-β1 and the α-subunits(s) of the G-protein(s) Gq and/or Gll. These studies show that testicular myoid and Sertoli cells use at least three G protein-coupled receptors (Vla-vasopressins, ETA-endothelin and P2U-purinergic) to signal through phosphoinositidase C activation, that ET-1 can activate multiple signalling pathways in myoid cells, and that the ET-1-stimulated phosphoinositidase C responses of myoid and Sertoli cells have different regulatory characteristics.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01076574
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