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Published in: Medical Oncology 10/2023

01-10-2023 | Insulins | Original Paper

Effect of insulin on IR and GLP1-R expressions in HT22 cells

Authors: Melek Tunc-Ata, Zeynep Mine Altunay, Aysel Alphan, Vural Kucukatay

Published in: Medical Oncology | Issue 10/2023

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Abstract

Insulin is a significant growth factor that specifically binds to the insulin receptor (IR) in the brain and then activates the PI3K-AKT pathway. Glucagon-like peptide 1 (GLP-1) has a variety of functions including neuroprotection, support for neurogenesis, and increasing insulin signal. This study aims to investigate the effect of insulin administered to immortalized clonal mouse hippocampal cell line (HT22) at different doses and intervals on IR, insulin receptor A (IRA), insulin receptor B (IRB), and Glucagon-like peptide 1 receptor (GLP1-R) mRNA expression and protein levels. The cells were planted in 6 well plates at a density of 3 × 105/4 × 105. Cells treated with insulin at different concentrations (5, 10, and 40 nM) were collected at 0.5, 2, 8, 16, and 24 h. RT-PCR and western blot analysis were used to measure mRNA expression and protein levels. Our results showed that insulin has short and long-term effects on IR and GLP1-R expression depending on dose and time. These findings may guide future studies targeting IR isoforms and GLP1-R in particular, as well as determining the optimal dose and duration of insulin stimulation in insulin signaling research.
Literature
2.
go back to reference Ullrich A, Bell JR, Chen EY, Herrera R, Petruzzelli LM, et al. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature. 1985;313:756–61.CrossRefPubMed Ullrich A, Bell JR, Chen EY, Herrera R, Petruzzelli LM, et al. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature. 1985;313:756–61.CrossRefPubMed
3.
go back to reference Frasca F, Pandini G, Scalia P, Sciacca L, Mineo R, et al. Insulin receptor isoform A, a newly recognized, high-affinity insulin-like growth factor II receptor in fetal and cancer cells. Mol Cell Biol. 1999;19:3278–88.CrossRefPubMedPubMedCentral Frasca F, Pandini G, Scalia P, Sciacca L, Mineo R, et al. Insulin receptor isoform A, a newly recognized, high-affinity insulin-like growth factor II receptor in fetal and cancer cells. Mol Cell Biol. 1999;19:3278–88.CrossRefPubMedPubMedCentral
4.
go back to reference Belfiore A, Frasca F, Pandini G, Sciacca L, Vigneri R. Insulin receptor isoforms and insulin receptor/insulin-like growth factor receptor hybrids in physiology and disease. Endocr Rev. 2009;30:586–623.CrossRefPubMed Belfiore A, Frasca F, Pandini G, Sciacca L, Vigneri R. Insulin receptor isoforms and insulin receptor/insulin-like growth factor receptor hybrids in physiology and disease. Endocr Rev. 2009;30:586–623.CrossRefPubMed
5.
6.
go back to reference van der Heide LP, Ramakers GMJ, Smidt MP. Insulin signaling in the central nervous system: learning to survive. Prog Neurobiol. 2006;79:205–21.CrossRefPubMed van der Heide LP, Ramakers GMJ, Smidt MP. Insulin signaling in the central nervous system: learning to survive. Prog Neurobiol. 2006;79:205–21.CrossRefPubMed
7.
go back to reference Heni M, Kullmann S, Preissl H, Fritsche A, Haring HU. Impaired insülin action in the human brain: causes and metabolic consequences. Nat Rev Endocrinol. 2015;11:701–11.CrossRefPubMed Heni M, Kullmann S, Preissl H, Fritsche A, Haring HU. Impaired insülin action in the human brain: causes and metabolic consequences. Nat Rev Endocrinol. 2015;11:701–11.CrossRefPubMed
8.
go back to reference Biessels GJ, Reagan LP. Hippocampal insulin resistance and cognitive dysfunction. Nat Rev Neurosci. 2015;16:660–71.CrossRefPubMed Biessels GJ, Reagan LP. Hippocampal insulin resistance and cognitive dysfunction. Nat Rev Neurosci. 2015;16:660–71.CrossRefPubMed
9.
go back to reference Messier C, Teutenberg K. The role of insulin, insulin growth factor, and insulin-degrading enzyme in brain aging and Alzheimer’s disease. Neural Plast. 2005;12:311–28.CrossRefPubMedPubMedCentral Messier C, Teutenberg K. The role of insulin, insulin growth factor, and insulin-degrading enzyme in brain aging and Alzheimer’s disease. Neural Plast. 2005;12:311–28.CrossRefPubMedPubMedCentral
10.
go back to reference Campbell JE, Drucker DJ. Pharmacology, physiology, and mechanisms of incretin hormone action. Cell Metab. 2013;17:819–37.CrossRefPubMed Campbell JE, Drucker DJ. Pharmacology, physiology, and mechanisms of incretin hormone action. Cell Metab. 2013;17:819–37.CrossRefPubMed
11.
go back to reference Madsbad S, Kielgast U, Asmar M, Deacon CF, Torekov SS, et al. An overview of once-weekly glucagon-like peptide-1 receptor agonists–available efficacy and safety data and perspectives for the future. Diabetes Obes Metab. 2011;13:394–407.CrossRefPubMed Madsbad S, Kielgast U, Asmar M, Deacon CF, Torekov SS, et al. An overview of once-weekly glucagon-like peptide-1 receptor agonists–available efficacy and safety data and perspectives for the future. Diabetes Obes Metab. 2011;13:394–407.CrossRefPubMed
12.
go back to reference Long-Smith CM, Manning S, McClean PL, Coakley MF, O’Halloran DJ, et al. The diabetes drug liraglutide ameliorates aberrant insulin receptor localization and signaling in parallel with decreasing both amyloid-beta plaque and glial pathology in a mouse model of Alzheimer’s disease. Neuromol Med. 2013;15:102–14.CrossRef Long-Smith CM, Manning S, McClean PL, Coakley MF, O’Halloran DJ, et al. The diabetes drug liraglutide ameliorates aberrant insulin receptor localization and signaling in parallel with decreasing both amyloid-beta plaque and glial pathology in a mouse model of Alzheimer’s disease. Neuromol Med. 2013;15:102–14.CrossRef
13.
go back to reference Zhou M, Chen S, Peng P, Gu Z, Yu J, et al. Dulaglutide ameliorates STZ-induced AD-like impairment of learning and memory ability by modulating hyperphosphorylation of tau and NFs through GSK3beta. Biochem Biophys Res Commun. 2019;511:154–60.CrossRefPubMed Zhou M, Chen S, Peng P, Gu Z, Yu J, et al. Dulaglutide ameliorates STZ-induced AD-like impairment of learning and memory ability by modulating hyperphosphorylation of tau and NFs through GSK3beta. Biochem Biophys Res Commun. 2019;511:154–60.CrossRefPubMed
14.
go back to reference Yang Y, Ma D, Xu W, Chen F, Du T, et al. Exendin-4 reduces tau hyperphosphorylation in type 2 diabetic rats via increasing brain insulin level. Mol Cell Neurosci. 2016;70:68–75.CrossRefPubMed Yang Y, Ma D, Xu W, Chen F, Du T, et al. Exendin-4 reduces tau hyperphosphorylation in type 2 diabetic rats via increasing brain insulin level. Mol Cell Neurosci. 2016;70:68–75.CrossRefPubMed
15.
go back to reference Khan M, Rutten BPF, Kim MO. MST1 regulates neuronal cell death via JNK/Casp3 signaling pathway in HFD mouse brain and HT22 cells. Int J Mol Sci. 2019;20:2504.CrossRefPubMedPubMedCentral Khan M, Rutten BPF, Kim MO. MST1 regulates neuronal cell death via JNK/Casp3 signaling pathway in HFD mouse brain and HT22 cells. Int J Mol Sci. 2019;20:2504.CrossRefPubMedPubMedCentral
16.
go back to reference Varshney P, Dey CS. P21-activated kinase 2 (PAK2) regulates glucose uptake and insülin sensitivity in neuronal cells. Mole Cell Endocrinol. 2016;429:50–61.CrossRef Varshney P, Dey CS. P21-activated kinase 2 (PAK2) regulates glucose uptake and insülin sensitivity in neuronal cells. Mole Cell Endocrinol. 2016;429:50–61.CrossRef
17.
go back to reference Li Y, Xiang Q, Yao YH, Liu JJ, Wang Y, et al. Inactivated AMPK- α2 promotes the progression of diabetic brain damage by Cdk5 phosphorylation at Thr485 site. Biochimie. 2020;168:277–84.CrossRefPubMed Li Y, Xiang Q, Yao YH, Liu JJ, Wang Y, et al. Inactivated AMPK- α2 promotes the progression of diabetic brain damage by Cdk5 phosphorylation at Thr485 site. Biochimie. 2020;168:277–84.CrossRefPubMed
18.
go back to reference Moruzzi N, Lazerri-Barcelo F, Valladolid-Acebes I, Moede T, Paschen M, et al. Tissue-specific expression of insulin receptor isoforms in obesity/type 2 diabetes mouse models. J Cell Mol Med. 2021;25:4800–13.CrossRefPubMedPubMedCentral Moruzzi N, Lazerri-Barcelo F, Valladolid-Acebes I, Moede T, Paschen M, et al. Tissue-specific expression of insulin receptor isoforms in obesity/type 2 diabetes mouse models. J Cell Mol Med. 2021;25:4800–13.CrossRefPubMedPubMedCentral
19.
go back to reference Zhu L, Zhou J, Pan Y, Lv J, Liu Y, et al. Glucagon-like peptide-1 receptor expression and its functions are regulated by androgen. Biomed Pharmacother. 2019;120: 109555.CrossRefPubMed Zhu L, Zhou J, Pan Y, Lv J, Liu Y, et al. Glucagon-like peptide-1 receptor expression and its functions are regulated by androgen. Biomed Pharmacother. 2019;120: 109555.CrossRefPubMed
20.
go back to reference Aoyama E, Watari I, Podyma-Inoue KA, Yanagishita M, Ono T. Expression of glucagon-like peptide-1 receptor and glucose-dependent insulinotropic polypeptide receptor is regulated by the glucose concentration in mouse osteoblastic MC3T3-E1 cells. Int J Mol Med. 2014;34:475–82.CrossRefPubMed Aoyama E, Watari I, Podyma-Inoue KA, Yanagishita M, Ono T. Expression of glucagon-like peptide-1 receptor and glucose-dependent insulinotropic polypeptide receptor is regulated by the glucose concentration in mouse osteoblastic MC3T3-E1 cells. Int J Mol Med. 2014;34:475–82.CrossRefPubMed
21.
go back to reference Apostolatos A, Song S, Acosta S, Peart M, Watson JE, et al. Insulin promotes neuronal survival via the alternatively spliced protein kinase CδII isoform. J Biol Chem. 2012;287:9299–310.CrossRefPubMedPubMedCentral Apostolatos A, Song S, Acosta S, Peart M, Watson JE, et al. Insulin promotes neuronal survival via the alternatively spliced protein kinase CδII isoform. J Biol Chem. 2012;287:9299–310.CrossRefPubMedPubMedCentral
22.
go back to reference Bassit GE, Patel RS, Carter G, Shibu V, Patel AA, et al. MALAT1 in human adipose stem cells modulates survival and alternative splicing of PKCδII in HT22 cells. Endocrinology. 2017;158:183–95.PubMed Bassit GE, Patel RS, Carter G, Shibu V, Patel AA, et al. MALAT1 in human adipose stem cells modulates survival and alternative splicing of PKCδII in HT22 cells. Endocrinology. 2017;158:183–95.PubMed
23.
go back to reference Bagaméry F, Varga K, Kecsmár K, Vincze I, Szöko E, et al. Lack of insulin resistance in response to streptozotocin treatment in neuronal SH-SY5Y cell line. J Neural Transm. 2020;127:71–80.CrossRefPubMed Bagaméry F, Varga K, Kecsmár K, Vincze I, Szöko E, et al. Lack of insulin resistance in response to streptozotocin treatment in neuronal SH-SY5Y cell line. J Neural Transm. 2020;127:71–80.CrossRefPubMed
24.
go back to reference Frazier HN, Ghoweri AO, Anderson KL, Lin RL, Popa GJ, et al. Elevating insulin signaling using a constitutively active insulin receptor increases glucose metabolism and expression of GLUT3 in hippocampal neurons. Front Neurosci. 2020;14:668.CrossRefPubMedPubMedCentral Frazier HN, Ghoweri AO, Anderson KL, Lin RL, Popa GJ, et al. Elevating insulin signaling using a constitutively active insulin receptor increases glucose metabolism and expression of GLUT3 in hippocampal neurons. Front Neurosci. 2020;14:668.CrossRefPubMedPubMedCentral
25.
go back to reference Koshal P, Kumar P. Neurochemical modulation involved in the beneficial effect of liraglutide, GLP-1 agonist on PTZ kindling epilepsy-induced comorbidities in mice. Mol Cell Biochem. 2016;415:77–87.CrossRefPubMed Koshal P, Kumar P. Neurochemical modulation involved in the beneficial effect of liraglutide, GLP-1 agonist on PTZ kindling epilepsy-induced comorbidities in mice. Mol Cell Biochem. 2016;415:77–87.CrossRefPubMed
26.
go back to reference Wen Y, Wu K, Xie Y, Dan W, Zhan Y, et al. Inhibitory effects of glucagon-like peptide-1 receptor on epilepsy. Biochem Biophys Res Commun. 2019;511:79–86.CrossRefPubMed Wen Y, Wu K, Xie Y, Dan W, Zhan Y, et al. Inhibitory effects of glucagon-like peptide-1 receptor on epilepsy. Biochem Biophys Res Commun. 2019;511:79–86.CrossRefPubMed
27.
go back to reference Hussein AM, Eldosoky M, El-Shafey M, El-Mesery M, Abbas KM, et al. Effects of GLP-1 receptor activation on a pentylenetetrazole-kindling rat model. Brain Sci. 2019;9:108.CrossRefPubMedPubMedCentral Hussein AM, Eldosoky M, El-Shafey M, El-Mesery M, Abbas KM, et al. Effects of GLP-1 receptor activation on a pentylenetetrazole-kindling rat model. Brain Sci. 2019;9:108.CrossRefPubMedPubMedCentral
28.
29.
go back to reference Liu Y, Beyer A, Aebersold R. On the dependency of cellular protein levels on mRNA abundance. Cell. 2016;165:535–50.CrossRefPubMed Liu Y, Beyer A, Aebersold R. On the dependency of cellular protein levels on mRNA abundance. Cell. 2016;165:535–50.CrossRefPubMed
Metadata
Title
Effect of insulin on IR and GLP1-R expressions in HT22 cells
Authors
Melek Tunc-Ata
Zeynep Mine Altunay
Aysel Alphan
Vural Kucukatay
Publication date
01-10-2023
Publisher
Springer US
Keywords
Insulins
Insulins
Published in
Medical Oncology / Issue 10/2023
Print ISSN: 1357-0560
Electronic ISSN: 1559-131X
DOI
https://doi.org/10.1007/s12032-023-02172-w

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