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 V1a-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)P2 hydrolysis [EC50 for endothelin-1 (ET-1) approximately 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 approximately 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)P2 hydrolysis. PtdIns(4,5)P2 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)P2 hydrolysis in the TM4 murine Sertoli cell line (EC50 approximately 0.6 nM), and this response was negatively regulated by protein kinase C activation. 5. No receptor-stimulated 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-beta 1 and the alpha-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 (V1a-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.

Adenosine Triphosphate↗

PubMedDate not supplied

Hormone-mediated inositol lipid breakdown in hepatocytes and WRK1 cells: relationship to receptor function.

All hormones and neurotransmitters which provoke their intracellular effects by increasing the cytosolic concentration of Ca2+ in their target cells also stimulate the breakdown of inositol phospholipids. Much evidence suggests that this breakdown is intimately involved in the mechanism which couples cell-surface receptor activation to intracellular Ca2+ mobilization. Recent results indicate that the primary, receptor-mediated event in stimulated cells is a phosphodiesteric hydrolysis of phosphatidylinositol 4,5-bisphosphate to yield inositol trisphosphate and diacylglycerol. It is likely that both products of this reaction fulfill 'second messenger' roles within stimulated cells.

Acetylcholine↗

PubMedDate not supplied

A vasopressin-like peptide in the mammalian sympathetic nervous system.

Vasopressin was among the first mammalian hormonal peptides to be identified and to have its structure determined. Its only undisputed physiological role is as a circulating neurohypophyseal antidiuretic hormone. Other notable effects of vasopressin on peripheral tissues include contraction of vascular smooth muscle, leading to elevation of blood pressure, and activation of glycogenolysis in liver. It has long been clear that vascular smooth muscle and hepatocytes are relatively insensitive to the low concentrations of vasopressin normally present in the circulation, and the physiological significance of their responses has therefore been in doubt. We now report that a new bioactive and immunoreactive vasopressin-like peptide (VLP) is widely distributed in the sympathetic nervous system of mammals, both in the principal noradrenergic neurones of ganglia and in nerve fibres innervating peripheral tissues. In addition to other peptides described in the mammalian sympathetic nervous system, VLP must be considered as a possible mediator of the non-adrenergic responses to sympathetic activation. Moreover, many of the effects previously attributed to circulating vasopressin may be neurally evoked.

Animals↗

PubMedDate not supplied

Stepwise enzymatic dephosphorylation of inositol 1,4,5-trisphosphate to inositol in liver.

Many receptors for hormones, neurotransmitters and other signals cause hydrolysis of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and effect a rise in cytosolic Ca2+ concentration. The inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) liberated during PtdIns(4,5)P2 breakdown seems to serve as a second messenger that activates the release of Ca2+ from a nonmitochondrial intracellular compartment. As expected if it is an important intracellular messenger, Ins(1,4,5)P3 is relatively rapidly degraded, both within stimulated cells and when added to homogenates of blowfly salivary gland or to permeabilized, but not intact, hepatocytes. Here we report that the dephosphorylation reactions responsible for the conversion of Ins(1,4,5)P3 to free inositol in rat liver are catalysed by two or more enzymes, and that these reactions are distributed between the plasma membrane and cytosol. The Ins(1,4,5)P3 5-phosphatase and inositol 1-phosphate (Ins(1)P) phosphatase of liver appear similar to enzymes described previously in erythrocytes and brain.

Animals↗

PubMedDate not supplied

The effect of advice to stop smoking on arterial disease patients, assessed by serum thiocyanate levels.

Serum thiocyanate has shown itself to be useful in distinguishing smokers from non-smokers. Our anti-smoking attitude resulted in 12 out of 32 smokers stopping smoking during the trial. Further, our evidence suggests that a patient who says he has given up smoking entirely can probably be believed, but a claim to have reduced smoking may be less reliable.

Adult↗