The Role of Neurotransmitters in Protection against Amyloid-β Toxicity by KiSS-1 Overexpression in SH-SY5Y Neurons

Abstract

Recent studies have suggested that the kisspeptin (KP) and kissorphin (KSO) peptides have neuroprotective actions against the Alzheimer's amyloid-β (Aβ) peptide. Overexpression of the human KiSS-1 gene that codes for KP and KSO peptides in SH-SY5Y neurons has also been shown to inhibit Aβ neurotoxicity. The in vivo actions of KP include activation of neuroendocrine and neurotransmitter systems. The present study used antagonists of KP, neuropeptide FF (NPFF), opioids, oxytocin, estrogen, adrenergic, cholinergic, dopaminergic, serotonergic, and γ-aminobutyric acid (GABA) receptors plus inhibitors of catalase, cyclooxygenase, nitric oxide synthase, and the mitogen activated protein kinase cascade to characterize the KiSS-1 gene overexpression neuroprotection against Aβ cell model. The results showed that KiSS-1 overexpression is neuroprotective against Aβ and the action appears to involve the KP or KSO peptide products of KiSS-1 processing. The mechanism of neuroprotection does not involve the activation of the KP or NPFF receptors. Opioids play a role in the toxicity of Aβ in the KiSS-1 overexpression system and opioid antagonists naloxone or naltrexone inhibited Aβ toxicity. The mechanism of KiSS-1 overexpression induced protection against Aβ appears to have an oxytocin plus a cyclooxygenase dependent component, with the oxytocin antagonist atosiban and the cyclooxygenase inhibitor SC-560 both enhancing the toxicity of Aβ.

Re-evaluation of Culture Condition of PC12 and SH-SY5Y Cells Based on Growth Rate and Amino Acid Consumption

“…We also found that treatment of amyloid peptides such as Aβ and Aβ 25-35 , nearly halved cell viability. The extent of sensitivity to amyloid peptides was similar to that of previously published values (9,11,12,13,15,18). These evidence suggests that the use of DMEM (supplemented with 10%FBS and antibiotics) is the best culture medium for both PC12 and SH-SY5Y cells.…”

“…As models of neurons, rat PC12 pheochromocytoma cells (6) and human SH-SY5Y neuroblastoma cells (7,8) have been used by many investigators to search for neuroprotective substances. However, the composition of the culture media they have used is quite different from researcher to researcher: RPMI1640 (differentiated PC12) (9, 10), RPMI (PC12) (11), DMEM (PC12) (12,13), DMEM (SH-SY5Y) (14), DMEM/Ham's F-12 (1:1) (SH-SY5Y) (15,16), MEM/F-12 (SY-SY5Y) (17) and DMEM/F-12 + non-essential amino acids (SH-SY5Y) (18), all supplemented with serum and antibiotics. It is not clear why the inclusion of F-12 and non-essential amino acids is necessary to culture SH-SY5Y cells.…”

Search of Neuroprotective Polyphenols Using the “Overlay” Isolation Method

“…Furthermore, it has recently been reported that neuronal protection caused by blackberry polyphenols is produced by mechanisms other than modulating reactive oxygen species (ROS) levels [ 52 ]. It is possible that both oxidative and other mechanisms, such as: Autophagy [ 21 ]; heat shock protein (HSP)-70 [ 22 ]; JAK2/STAT5/Bcl-xL [ 24 ]; phosphor HSP-20 [ 28 ]; amyloid disaggregation [ 29 ]; and, the overexpression of the KiSS gene [ 30 ], may be involved in the induction of neurotoxicity (column D in Table 1 ).…”

“…However, there was no uniformity in the culture condition of PC12 and SH-SY5Y cells in previous investigations. The culture media used was: Dulbecco’s modified Eagle’s medium (DMEM); the minimum essential medium (MEM); RPMI1640; a mixture of DMEM and Ham′s F12 (1:1); and, non-essential amino acids (NEAA), which were supplemented with fetal bovine serum (FBS), alone or together with, horse serum (HS) (column A in Table 1 ) [ 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 ]. Differentiated and undifferentiated cells (column B), inoculated at different cell densities, were exposed to considerably different concentrations of neurotoxic agents (column C) in uncoated, collagen, or poly-lysine-coated plates ( Table 1 ).…”

Bioinformatics analyses of significant genes, related pathways and candidate prognostic biomarkers in Alzheimer’s disease

“…GABRA6 [64], GABRA1 [65], GABRG2 [66], SLC4A10 [67], HCN1 [68], GABRA4 [69], KCNC2 [70], SCN2A [71], SYN2 [72], FGF12 [73], SCN8A [74], OLFM3 [75], PLCB1 [76], KCNQ5 [77], TUBB2A [78], SIK1 [79], ABCC2 [80], SLC6A12 [81] and COL6A2 [82] have been reported to be closely related to the occurrence and development of epilepsy, but these genes might be novel target for AD. Previous investigation demonstrates that RGS4 [83], CXCL11 [84], EGR1 [85], CALB1 [86], BDNF (brain derived neurotrophic factor) [87], TERT (telomerase reverse transcriptase) [88], NEFL (neurofilament light) [89], SNAP25 [90], RPH3A [91], NRN1 [92], SYT1 [93], GRIN2B [94], AVP (arginine vasopressin) [95], VSNL1 [96], HTR2A [97], PAK3 [98], STXBP5L [99], HCRTR2 [100], SYP (synaptophysin) [101], SYT10 [102], PRKCE (protein kinase C epsilon) [103], NRG1 [104], KISS1 [105], NRXN3 [106], RAB3A [107], IGF1 [108], PLK2 [109], CBLN4 [110], CAP2 [111], SV2B [112], CAMK4 [113], INA (internexin neuronal intermediate filament protein alpha) [114], GAP43 [115], TTR (transthyretin) [116], CXCR2 [117], IL1R2 [118], CXCR4 [119], CCR2 [120], MYOCD (myocardin) [121], S100A12 [122], CXCR3 [123], PROK2 [124], CXCL8 [125], RGS1 [126], NOTCH3 [127], P2RX7 [128], NGFR (nerve growth factor receptor) [129], GDF15 [130], CR1 [131], ADORA2A [132], GPER1 [133], FCGR2B [134], MMP9 [135], CNN2 [136...…”