Isolation of a murine glucose-dependent insulinotropic polypeptide (GIP) cDNA from a tumor cell line (STC6–14) and quantification of glucose-induced increases in GIP mRNA

Schieldrop, Phil J.; Gelling, Richard W.; Elliot, Ruan; Hewitt, Jeff; Kieffer, Timothy J.; McIntosh, Christopher H.S.; Pederson, Raymond A.

doi:10.1016/0167-4781(96)00103-0

Cited by 7 publications

(2 citation statements)

References 7 publications

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“…b-actin was amplified for 20 cycles using published primers and cycling conditions (Riechmann et al, 1994). Primers were designed to amplify nucleotides 110-471 of the coding sequence for mouse GIP mRNA (Schieldrop et al, 1996). They are (5 0 )-CTC GAC CTC GAG GTC CAA GG-(3 0 ) and (5 0 )-AGG TCA GAG TCC AGT CCT GC-(3 0 ), respectively.…”

Section: Rt-pcr Assaysmentioning

confidence: 99%

Novel insulin/GIP co‐producing cell lines provide unexpected insights into Gut K‐cell function in vivo

Ramshur

Rull

Wice

2002

Journal Cellular Physiology

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Enteroendocrine (EE) cells represent complex, rare, and diffusely-distributed intestinal epithelial cells making them difficult to study in vivo. A specific sub-population of EE cells called Gut K-cells produces and secretes glucose-dependent insulinotropic peptide (GIP), a hormone important for glucose homeostasis. The factors that regulate hormone production and secretion, as well as the timing of peptide release, are remarkably similar for K-cells and islet beta-cells suggesting engineering insulin production by K-cells is a potential gene therapeutic strategy to treat diabetes. K-cell lines could be used to study the feasibility of this potential therapy and to understand Gut K-cell physiology in general. Heterogeneous STC-1 cells were transfected with a plasmid (pGIP/Neo) encoding neomycin phosphotransferase, driven by the GIP promoter-only cells in which the GIP promoter was active survived genetic selection. Additional clones expressing pGIP/Neo plus a GIP promoter/insulin transgene were isolated-only doubly transfected cells produced preproinsulin mRNA. Bioactive insulin was stored and then released following stimulation with arginine, peptones, and bombesin-physiological GIP secretagogues. Like K-cells in vivo, the GIP/insulin-producing cells express the critical glucose sensing enzyme, glucokinase. However, glucose did not regulate insulin or GIP secretion or mRNA levels. Conversely, glyceraldehyde and methyl-pyruvate were secretagogues, indicating cells depolarized in response to changes in intracellular metabolite levels. Potassium channel opening drugs and sulphonylureas had little effect on insulin secretion by K-cells. The K-cell lines also express relatively low levels of Kir 6.1, Kir 6.2, SUR1, and SUR2 suggesting secretion is independent of K(ATP) channels. These results provided unexpected insights into K-cell physiology and our experimental strategy could be easily modified to isolate/characterize additional EE cell populations.

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Section: Rt-pcr Assaysmentioning

confidence: 99%

Novel insulin/GIP co‐producing cell lines provide unexpected insights into Gut K‐cell function in vivo

Ramshur

Rull

Wice

2002

Journal Cellular Physiology

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“…PreproGIP mRNA Expression-The GIP precursor is a protein of 144 (rodent) or 153 (human) amino acids (33)(34)(35) expressed in the endocrine intestinal K cells that exhibits the highest density in the proximal jejunum but is found in the entire intestinal mucosa (17). Normal processing is believed to depend on cleavage at the conserved sites 19 RGPR 22 2 at the N terminus and 65 R2EAR 68 at the C terminus, resulting in the biologically active peptide GIP (proGIP-(23-64)) and an N-terminal and C-terminal fragment (Fig.…”

Section: Structure Of the Gip Precursor Gip And Pc1/3 Co-localizatiomentioning

confidence: 99%

Prohormone Convertase 1/3 Is Essential for Processing of the Glucose-dependent Insulinotropic Polypeptide Precursor

Ugleholdt

Poulsen

Holst

et al. 2006

Journal of Biological Chemistry

100

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Glucose-Dependent Insulinotropic Polypeptide (GIP)

Wolfe¹,

Boylan²,

Kieffer³

et al. 1999

Gastrointestinal Endocrinology

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Isolation of a murine glucose-dependent insulinotropic polypeptide (GIP) cDNA from a tumor cell line (STC6–14) and quantification of glucose-induced increases in GIP mRNA

Cited by 7 publications

References 7 publications

Novel insulin/GIP co‐producing cell lines provide unexpected insights into Gut K‐cell function in vivo

Novel insulin/GIP co‐producing cell lines provide unexpected insights into Gut K‐cell function in vivo

Prohormone Convertase 1/3 Is Essential for Processing of the Glucose-dependent Insulinotropic Polypeptide Precursor

Glucose-Dependent Insulinotropic Polypeptide (GIP)

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