Numerous studies recently showed that the inhibitory neurotransmitter, γ-aminobutyric acid (GABA), can stimulate cerebral angiogenesis and promote neurovascular coupling by activating the ionotropic GABA_A receptors on cerebrovascular endothelial cells, whereas the endothelial role of the metabotropic GABA_B receptors is still unknown. Preliminary evidence showed that GABA_A receptor stimulation can induce an increase in endothelial Ca²⁺ levels, but the underlying signaling pathway remains to be fully unraveled. In the present investigation, we found that GABA evoked a biphasic elevation in [Ca²⁺]_i that was initiated by inositol-1,4,5-trisphosphate- and nicotinic acid adenine dinucleotide phosphate-dependent Ca²⁺ release from neutral and acidic Ca²⁺ stores, respectively, and sustained by store-operated Ca²⁺ entry. GABA_A and GABA_B receptors were both required to trigger the endothelial Ca²⁺ response. Unexpectedly, we found that the GABA_A receptors signal in a flux-independent manner via the metabotropic GABA_B receptors. Likewise, the full Ca²⁺ response to GABA_B receptors requires functional GABA_A receptors. This study, therefore, sheds novel light on the molecular mechanisms by which GABA controls endothelial signaling at the neurovascular unit.

Background: Platelets can support cancer progression via the release of microparticles and microvesicles that enhance the migratory behaviour of recipient cancer cells. We recently showed that platelet-derived extracellular vesicles (PEVs) stimulate migration and invasiveness in highly metastatic MDA-MB-231 cells by stimulating the phosphorylation of p38 MAPK and the myosin light chain 2 (MLC2). Herein, we assessed whether the pro-migratory effect of PEVs involves the remodelling of the Ca²⁺ handling machinery, which drives MDA-MB-231 cell motility. Methods: PEVs were isolated from human blood platelets, and Fura-2/AM Ca²⁺ imaging, RT-qPCR, and immunoblotting were exploited to assess their effect on intracellular Ca²⁺ dynamics and Ca²⁺-dependent migratory processes in MDA-MB-231 cells. Results: Pretreating MDA-MB-231 cells with PEVs for 24 h caused an increase in Ca²⁺ release from the endoplasmic reticulum (ER) due to the up-regulation of SERCA2B and InsP₃R1/InsP₃R2 mRNAs and proteins. The consequent enhancement of ER Ca²⁺ depletion led to a significant increase in store-operated Ca²⁺ entry. The larger Ca²⁺ mobilization from the ER was required to potentiate serum-induced migration by recruiting p38 MAPK and MLC2. Conclusions: PEVs stimulate migration in the highly metastatic MDA-MB-231 breast cancer cell line by inducing a partial remodelling of the Ca²⁺ handling machinery.

Exogenous administration of hydrogen sulfide (H₂S) is emerging as an alternative anticancer treatment. H₂S-releasing compounds have been shown to exert a strong anticancer effect by suppressing proliferation and/or inducing apoptosis in several cancer cell types, including colorectal carcinoma (CRC). The mechanism whereby exogenous H₂S affects CRC cell proliferation is yet to be clearly elucidated, but it could involve an increase in intracellular Ca²⁺ concentration ([Ca²⁺]_i). Herein, we sought to assess for the first time whether (and how) sodium hydrosulfide (NaHS), one of the most widely employed H₂S donors, induced intracellular Ca²⁺ signals in primary cultures of human metastatic CRC (mCRC) cells. We provided the evidence that NaHS induced extracellular Ca²⁺ entry in mCRC cells by activating the Ca²⁺-permeable channel Transient Receptor Potential Vanilloid 1 (TRPV1) followed by the Na⁺-dependent recruitment of the reverse-mode of the Na⁺/Ca²⁺ (NCX) exchanger. In agreement with these observations, TRPV1 protein was expressed and capsaicin, a selective TRPV1 agonist, induced Ca²⁺ influx by engaging both TRPV1 and NCX in mCRC cells. Finally, NaHS reduced mCRC cell proliferation, but did not promote apoptosis or aberrant mitochondrial depolarization. These data support the notion that exogenous administration of H₂S may prevent mCRC cell proliferation through an increase in [Ca²⁺]_i, which is triggered by TRPV1.