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Nicotinic acid adenine dinucleotide phosphate (NAADP) is a potent Ca2؉ -mobilizing messenger that in many cells releases Ca 2؉ from the endolysosomal system. Recent studies have shown that NAADP-induced Ca 2؉ mobilization is mediated by the two-pore channels (TPCs). Whether NAADP acts as a messenger in astrocytes is unclear, and downstream functional consequences have yet to be defined. Here, we show that intracellular delivery of NAADP evokes Ca 2؉ signals from acidic organelles in rat astrocytes and that these signals are potentiated upon overexpression of TPCs. We also show that NAADP increases acidic vesicular organelle formation and levels of the autophagic markers, LC3II and beclin-1. NAADP-mediated increases in LC3II levels were reduced in cells expressing a dominant-negative TPC2 construct. Our data provide evidence that NAADP-evoked Ca 2؉ signals mediated by TPCs regulate autophagy. Increases in cytosolic Ca2ϩ regulate a myriad of cellular functions including information processing in the central nervous system (1). In many cells, these increases can be driven by mobilization of intracellular Ca 2ϩ stores (2). Much attention has focused on the endoplasmic reticulum as a Ca 2ϩ store (3), but accumulating evidence also implicates acidic organelles such as lysosomes in the control of Ca 2ϩ dynamics (4). In particular, NAADP 2 has emerged as a novel intracellular Ca 2ϩ -mobilizing messenger that links cell surface stimulation to the release of Ca 2ϩ from acidic Ca 2ϩ stores (5). Changes in the concentration of cytosolic Ca 2ϩ in glial cells are key for bidirectional control of neuronal activity (1). Previous studies have shown that extracellular application of NAADP can evoke Ca 2ϩ signals in astrocytes, consistent with a messenger role for NAADP in this cell type, following its internalization (6). Interpretation of these results, however, is clouded by the demonstrated lack of specificity with respect to related nucleotides (6) and by the potential activation by NAADP of cell surface purinergic receptors (7). Whether NAADP acts as an intracellular messenger in astrocytes is therefore unclear.Although the role of inositol trisphosphate and ryanodine receptors is established in mediating Ca 2ϩ release from the ER in response to inositol trisphosphate and cyclic ADP-ribose, respectively (2), the molecular basis for Ca 2ϩ release by NAADP from acidic organelles is less certain (8). In a series of recent studies, however, a novel family of Ca 2ϩ channels, known as the two-pore channels (TPCs), have emerged as likely targets (9). Thus, TPCs localize to endosomes and/or lysosomes through an identified targeting motif and enhance NAADPmediated cytosolic Ca 2ϩ signals when overexpressed (10 -12). Inhibition of TPC expression/function using siRNA (10), TPC knock-out mice (11), or a dominant-negative TPC construct (10) reduces NAADP-evoked Ca 2ϩ signals, and biophysical analyses indicate that TPCs are NAADP-gated Ca 2ϩ -permeable channels (13). Moreover, a functional role for TPCs has been identified in events such ...

The mechanisms that regulate programmed cell death, such as apoptosis, and the cellular "self-eating" phenomenon of autophagy, share many regulatory systems and common pathways. These mechanisms have been extensively investigated over the last few years. Some intracellular structures may determine and control the autophagic fate of the cell such as the endoplasmic reticulum, mitochondria, and lysosomes. The coordination and interrelation of these organelles are crucial in maintaining calcium levels and general cellular homeostasis, as well as in regulating cell life and death under physiological and pathological conditions, including cancer, neurodegeneration, and aging. In this review, we discuss the crosstalk between the aforementioned organelles and their influence in apoptotic and autophagic processes.

Although AraC anti-leukaemia effects could be potentiated by 3MA autophagy inhibition after 24 h of exposure, leukaemia cell resistance, the main causes of treatment failure, is also promoted by autophagy initial stage impairment by 3MA, denoting the complex role of autophagy in leukaemia cells’ response to chemotherapy.