Multiple Pathways for High Voltage-Activated Ca2+ Influx in Anterior Pituitary Lactotrophs and Somatotrophs

作者：Tzour A; Sosial E; Meir T; Canello T; Naveh Many T; Gabizon R; Nussinovitch I^*

来源：Journal of Neuroendocrinology, 2013, 25(1): 76-86.

DOI：10.1111/j.1365-2826.2012.02372.x

• Ca2+ channels
• anterior pituitary cells
• somatotrophs
• lactotrophs
• lipid rafts

The present study demonstrates that a significant proportion of high voltage-activated (HVA) Ca2+ influx in native rat anterior pituitary cells is carried through non-L-type Ca2+ channels. Using whole-cell patch-clamp recordings and specific Ca2+ channel toxin blockers, we show that approximately 35% of the HVA Ca2+ influx in somatotrophs and lactotrophs is carried through Ca(v)2.1, Ca(v)2.2 and Ca(v)2.3 channels, and that somatotrophs and lactotrophs share similar proportions of these non-L-type Ca2+ channels. Furthermore, experiments on mixed populations of native anterior pituitary cells revealed that the fraction of HVA Ca2+ influx carried through these non-L-type Ca2+ channels might even be higher (approximately 46%), suggesting that non-L-type channels exist in the majority of native anterior pituitary cells. Using western blotting, immunoblots for alpha(1C), alpha(1D), alpha(1A), alpha(1B) and alpha(1E) Ca2+ channel subunits were identified in native rat anterior pituitary cells. Additionally, using reverse transcriptase-polymerase chain reaction, cDNA transcripts for alpha(1C), alpha(1D), alpha(1A) and alpha(1B) Ca2+ channel subunits were identified. Transcripts for alpha(1E) were nonspecific and transcripts for alpha(1S) were not detected at all (control). Taken together, these results clearly demonstrate the existence of multiple HVA Ca2+ channels in the membrane of rat native anterior pituitary cells. Whether these channels are segregated among different membrane compartments was investigated further in flotation assays, demonstrating that Ca(v)2.1, Ca(v)1.2 and caveolin-1 were mostly localised in light fractions of Nycodenz gradients (i.e. in lipid raft domains). Ca(v)1.3 channels were distributed among both light and heavy fractions of the gradients (i.e. among raft and nonraft domains), whereas Ca(v)2.2 and Ca(v)2.3 channels were distributed mostly among nonraft domains. In summary, in the present study, we demonstrate multiple pathways for HVA Ca2+ influx through L-type and non-L-type Ca2+ channels in the membrane of native anterior pituitary cells. The compartmentalisation of these channels among raft and nonraft membrane domains might be essential for their proper regulation by separate receptors and signalling pathways.

• 出版日期2013-1
• 单位河北医科大学