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Inositol lipid-mediated signalling in response to endothelin and ATP in the mammalian testis

  • Chapter
Signal Transduction Mechanisms

Part of the book series: Developments in Molecular and Cellular Biochemistry ((DMCB,volume 15))

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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 A1F4 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-βl and the a-subunits(s) of the G-protein(s) Gq and/or G11. 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.

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Abbreviations

PtdIns(4,5)P 2 :

phosphatidylinositol 4,5-bisphosphate

Ins, Insp, InsP 2, InsP 3, InsP 4, InsP 5, InsP 6 (and equivalent abbreviations with added locants):

myoinositol and myoinositol phosphates numbered by reference to D-myoinositol 1 - phosphate as InslP (see Biochemical Journal (1989) 258, 1-2]

ET-1:

endothelin-1

ET-3:

endothelin-3; PKC - protein kinase C

PDB:

phorbol 12,13-dibutyrate

IBMX:

isobutylmethylxanthine

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  1. Simon A. Rudge

    Present address: Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY, 11794-8651, USA

  2. Graham R. Brown

    Present address: Department of Molecular Medicine, Karolinska Institute, S-17176, Stockholm, Sweden

Authors and Affiliations

  1. Centre for Clinical Research in Immunology and Signalling and School of Biochemistry, University of Birmingham, Birmingham, B15 2TT, UK

    Simon A. Rudge, Phillip J. Hughes, Graham R. Brown, Robert H. Michell & Christopher J. Kirk

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Editors and Affiliations

  1. Department of Biochemistry, University of West Indies, St. Augustine, Trinidad, West Indies, Caribbean

    Junor A. Barnes  & Haldane G. Coore  & 

  2. Department of Geriatric Medicine, Karolinska Institute, Stockholm, Sweden

    Abdul H. Mohammed

  3. Department of Pathology College of Medicine Saskatoon Cancer Center, University of Saskatoon, Saskatoon, Saskatchewan, Canada

    Rajendra K. Sharma

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Rudge, S.A., Hughes, P.J., Brown, G.R., Michell, R.H., Kirk, C.J. (1995). Inositol lipid-mediated signalling in response to endothelin and ATP in the mammalian testis. In: Barnes, J.A., Coore, H.G., Mohammed, A.H., Sharma, R.K. (eds) Signal Transduction Mechanisms. Developments in Molecular and Cellular Biochemistry, vol 15. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2015-3_17

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