Prolonging the circulatory retention of SPIONs using dextran sulfate : in vivo tracking achieved by functionalisation with near-infrared dyes

<p>Intraperitoneal (Panels <b>A</b> and <b>C</b>) and oral (Panel <b>B</b>) glucose tolerance tests were performed on 6 hr fasted conscious C57BL/6J WT (closed symbols) and <i>Slc30a8</i> KO (open symbols) male mice as described in Materials and Methods. The IPGTT results in Panel <b>A</b> show the mean glucose concentrations ± S.E.M. in WT (n = 11; mean age ∼20 weeks) and <i>Slc30a8</i> KO (n = 9; mean age ∼20 weeks) animals. The OGTT results in Panel <b>B</b> show the mean glucose concentrations ± S.E.M. in WT (n = 16; mean age ∼22 weeks) and <i>Slc30a8</i> KO (n = 9; mean age ∼22 weeks) animals. The IPGTT results in Panel <b>C</b> show the mean glucose concentrations ± S.E.M. in WT (n = 17; mean age ∼4 weeks) and <i>Slc30a8</i> KO (n = 19; mean age ∼4 weeks) animals. *p<0.05 <i>versus</i> WT.</p

Analysis of insulin sensitivity in female C57BL/6J <i>Slc30a8</i> KO mice <i>in vivo</i>.

<p>Insulin tolerance tests were performed on 5 hr fasted conscious C57BL/6J WT (closed symbols) and <i>Slc30a8</i> KO (open symbols) female mice as described in Materials and Methods. Results show the mean glucose concentrations ± S.E.M. in WT (n = 8; mean age ∼18 weeks) and <i>Slc30a8</i> KO (n = 8; mean age ∼18 weeks) animals. In these groups of mice the initial glucose concentration at time zero was higher in KO mice (142.6±6.1 vs 120.8±4.9 mg/dl; p<0.02).</p

Analysis of insulin content and glucose-stimulated insulin secretion in male C57BL/6J <i>Slc30a8</i> KO mouse islets <i>in situ</i>.

<p>Islets were isolated from male C57BL/6J WT and <i>Slc30a8</i> KO mice and then insulin content (<b>Panel A</b>) and glucose-stimulated insulin secretion (<b>Panel B</b>) were assayed as described in Materials and Methods. Results show the mean data ± S.E.M. from 3 islet preparations isolated from ∼18 week old male mice. *p<0.05 <i>versus</i> 5 mM glucose.</p

Phenotypic characterization of high fat fed male C57BL/6J <i>Slc30a8</i> KO mice.

<p>Male WT (n = 5; mean age 44.6 weeks), heterozygous (HET) (n = 14; mean age 39.6 weeks) and KO (n = 6; mean age 49.5 weeks) mice were fed a high fat diet with body weights measured weekly (<b>Panel A</b>). After 8 weeks mice fasted for 4 hours and then weighed. One hour later mice were anesthetized and blood was isolated. Blood glucose (<b>Panel B</b>) and plasma cholesterol (<b>Panel C</b>) and insulin (<b>Panel D</b>) levels were determined as described in Materials and Methods. One week later IPGTTs were performed on 6 hr fasted conscious C57BL/6J WT (closed symbols) and <i>Slc30a8</i> KO (open symbols) male mice as described in Materials and Methods (Panel <b>E</b>). Results represent mean data ± S.E.M. *p<0.05 <i>versus</i> WT.</p

Normalized expression of selected <i>SLC30</i> genes by quantitative RT-PCR in 9–23 week old human fetal pancreas and adult human islets.

<p>Data in triplicate (mean ± S.E.M.) is normalized to endogenous <i>GAPDH</i> and quantified and expressed relative to 9 week old fetal samples. <i>SLC30A2</i> expression was detectable in the fetal pancreas from 19 weeks. *p<0.05 from 9 weeks for <i>SLC30A8</i> (black bars).</p

Phenotypic characterization of fasted C57BL/6J <i>Slc30a8</i> KO mice.

<p>Length: *F WT vs. F KO, p<0.01; Cholesterol: *M WT vs. M Het p<0.01; Insulin: *F WT vs. F Het, p<0.001; **F WT vs. F KO, p<0.05; Proinsulin: *M WT vs. KO, p<0.05.</p><p>At 16 weeks of age mice were fasted for 5 hours and then weighed. One hour later mice were anesthetized, their length was measured and blood isolated. Blood glucose and plasma cholesterol, triacylglycerol, glycerol, insulin and glucagon levels were determined as described in Materials and Methods. Results represent mean data ± S.E.M. obtained from the indicated number of animals in parentheses.</p

Biochemical characterization of C57BL/6J <i>Slc30a8</i> KO mice.

<p><b>Panel A:</b> Immunohistochemical staining of wild type and <i>Slc30a8</i> KO mouse pancreas with antisera raised to insulin, glucagon, and ZnT-8 was performed as described in Materials and Methods. Representative pictures are shown. WT, wild type; KO, knockout. <b>Panel B:</b> Zinc content in isolated islets was determined as described in Material and Methods. Results represent the mean ± S.E.M. (n = 3 independent islet preparations for each genotype with each islet preparation assayed in quintuplicate). *p<0.05 <i>versus</i> WT.</p

Analysis of insulin content and GSIS in female C57BL/6J <i>Slc30a8</i> KO mouse islets <i>in situ</i>.

<p>Islets were isolated from female WT and <i>Slc30a8</i> KO mice and then insulin content (<b>Panel A</b>) and GSIS (<b>Panel B</b>) were assayed as described in Materials and Methods. Results show the mean data ± S.E.M. from 6 islet preparations isolated from ∼18 week old female mice. *p<0.05 <i>versus</i> 5 mM glucose.</p