Psychometric evaluation of the Menstrual Joy Questionnaire.

Analysis of 10 terms in the Menstrual Joy Questionnaire of Delaney, Lupton, and Toth indicated that the 34 undergraduate students did not agree on the definitions of scale items. The authors discuss the use of this questionnaire as a stimulus in experimental research and as a measure of positive perimenstrual experience.

Photoaffinity labeling of muscle-type nicotinic acetylcholine receptors and neuronal/nicotinic alpha-bungarotoxin binding sites with a derivative of alpha-bungarotoxin.

Neuronal/nicotinic alpha-bungarotoxin binding sites (nBgtS) found in the nervous system are not well characterized. In this study, photolabile toxin derivatives have been used in affinity labeling protocols to investigate the subunit composition of nBgtS expressed by different neuron-like cell lines. Data obtained was compared to the known subunit composition of toxin-binding muscle-type nicotinic acetylcholine receptors (nAChR). Muscle-type nAChR-rich membranes prepared from Torpedo electroplax contain components with corrected apparent molecular sizes of 41, 46, 50, 62 and 66 kDa that are reactive with toxin. The photoaffinity labeling patterns for preparations derived from cells of the TE671 clone, which express muscle-type nAChR, are very similar to that of cells of the IMR-32 or SH-SY5Y clonal lines, which express nBgtS. There is consistent labeling of four polypeptides with corrected apparent molecular weights of 40, 43, 47 and 56 kDa. These results suggest that both mammalian muscle-type nAChR and mammalian nBgtS are similarly composed of at least four kinds of subunits.

Morphological and biochemical differentiation of the human medulloblastoma cell line TE671.

Cells of the human medulloblastoma clonal line TE671 exhibit polymorphism when grown in vitro in serum-supplemented medium. Under these conditions, cell numbers double every 18 h during log phase growth. These tumorigenic precursors of cerebellar interneurons are not contact-inhibited and approach densities of one million cells per cm2. TE671 cells in proliferative growth express a class of nicotinic acetylcholine receptors that are fully sensitive to functional blockade by the neurotoxin alpha-bungarotoxin (Bgt). TE671 cells grown in medium containing dibutyryl cyclic adenosine monophosphate (dbcAMP) rapidly undergo a distinctive morphological transformation characterized by neurite extension and formation of cell-cell contacts. The rate of cell division and cell saturation densities are diminished coordinately with these treatments. Sodium fluoride and forskolin induce similar changes in cell division and morphology as does dbcAMP, and these effects are potentiated by aluminum and the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, respectively. The high-affinity binding of Bgt to TE671 cells also is reduced on exposure to dbcAMP in a time and dose-dependent manner. The results suggest that activation of adenylate cyclase and the concomitant elevation of intracellular cAMP levels may be involved in the morphological transformation of TE671 cells to a mature, neuronal phenotype and in changes in the level of expression of a subtype of human neuronal nicotinic receptors. These studies establish a unique, neural tube-derived model system for investigation of the mechanisms involved in these processes.

Nicotinic agonists regulate alpha-bungarotoxin binding sites of TE671 human medulloblastoma cells.

The TE671 human medulloblastoma cell line expresses a variety of characteristics of human neurons. Among these characteristics is the expression of membrane-bound high-affinity binding sites for alpha-bungarotoxin, which is a potent antagonist of functional nicotinic acetylcholine receptors on these cells. These toxin binding sites represent a class of nicotinic receptor isotypes present in mammalian brain. Treatment of TE671 cells during proliferative growth phase with nicotine or carbamylcholine, but not with muscarine or d-tubocurarine, induced up to a five-fold increase in the density of radiolabeled toxin binding sites in crude membrane fractions. This effect was blocked by co-incubation with the nicotinic antagonists d-tubocurarine and decamethonium, but not by mecamylamine or by muscarinic antagonists. Following a 10-13 h lag phase upon removal of agonist, recovery of the up-regulated sites to control values occurred within an additional 10-20 h. These studies indicate that the expression of functional nicotinic acetylcholine receptors on TE671 cells is subject to regulation by nicotinic agonists. Studies of the murine CNS have consistently indicated nicotine-induced up-regulation of nicotinic acetylcholine receptors, thereby supporting the identification of the toxin binding site on these cells as the functional nicotinic receptor. Although a mechanism for this effect is not apparent, nicotine-induced receptor blockade does not appear to be involved.

Allosteric modification of alpha-bungarotoxin binding by the 'calcium channel antagonist' verapamil.

The putative calcium channel antagonist verapamil has been shown to affect a variety of other ion channels as well as neurotransmitter receptors. We report that verapamil reversibly inhibits binding of the nicotinic receptor probe 125I-alpha-bungarotoxin to membranes from rat brain, Torpedo californica, the rat pheochromocytoma cell line PC12 and the human medulloblastoma cell line TE671, with Ki values of 24.0, 90.5, 165.4 and 869.6 microM, respectively. These effects are calcium independent and insensitive to a variety of organic and inorganic calcium channel antagonists. While Hill coefficients suggest mechanistic differences for verapamil action in the tissues examined, derivative graphical analysis demonstrates a common mechanism of heterotypic, allosteric inhibition. Verapamil may be useful as a probe to elucidate aspects of receptor-effector coupling.