11 research outputs found

    Primer Sequences for Genes Quantified by Real-Time PCR<sup>a</sup>.

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    <p>Primer Sequences for Genes Quantified by Real-Time PCR<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162432#t002fn001" target="_blank"><sup>a</sup></a>.</p

    Mammary gland mRNA and protein expression of various serotonergic and immune markers is altered in response to high fat diet and <i>Tph1</i> deficiency.

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    <p>Wild type or <i>Tph1</i> deficient dams were either fed a high fat or low fat diet throughout pregnancy and lactation. Shown is (<i>A</i>) mammary gland mRNA expression of tryptophan hydroxylase 1, (<i>B</i>) total mammary gland protein expression of tryptophan hydroxylase 1, (<i>C</i>) mammary gland mRNA expression of nitric oxide synthase, (<i>D</i>) mammary gland mRNA expression of C-X-C motif chemokine 5, (<i>E</i>) mammary gland mRNA expression of tumor necrosis factor alpha. Values are means ± SEMs. Stars indicate statistical significance between groups (* = 0.05<<i>P</i><0.01, ** = 0.01<<i>P</i><0.001, *** = 0.001<<i>P</i><0.0001, **** = <i>P</i><0.0001). WT, wild type; <i>Tph1</i><sup><i>-/-</i></sup>, <i>Tph1</i> deficient; LFD, low fat diet; HFD, high fat diet.</p

    All dams fed a high fat diet weighed more than those fed a low fat diet, but only wild type high fat diet fed dams killed more pups at the onset of lactation.

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    <p>Wild type or <i>Tph1</i> deficient dams were either fed a high fat or low fat diet throughout pregnancy and lactation. Shown in (<i>A</i>) are the effects on dam weight and in (<i>B</i>) on pup mortality. Values are means ± SEMs. Stars indicate statistical significance between groups (* = 0.05<<i>P</i><0.01, ** = 0.01<<i>P</i><0.001, *** = 0.001<<i>P</i><0.0001). WT, wild type; <i>Tph1</i><sup><i>-/-</i></sup>, <i>Tph1</i> deficient; LFD, low fat diet; HFD, high fat diet; P0, day 0 of pregnancy; P7, day 7 of pregnancy; P17-20, day 17 to 20 of pregnancy; L0, day 0 of lactation; L10, day 10 of lactation.</p

    Milk fatty acid composition and mRNA expression of fat-related genes were altered in response to high fat or low fat diet fed to wild type or <i>Tph1</i> deficient dams.

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    <p>Wild type or <i>Tph1</i> deficient dams were either fed a high fat or low fat diet throughout pregnancy and lactation. Shown are the effects on (<i>A</i>) relative abundance of various fatty acids in milk from early lactation, (<i>B</i>) relative abundance of various fatty acids in milk on day 10 of lactation, (<i>C</i>) mammary gland mRNA expression of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3, (<i>D</i>) mammary gland mRNA expression of insulin-like growth factor binding protein 5, (<i>E</i>) mammary gland mRNA expression of cholecystekinin. Values are means ± SEMs. Stars indicate statistical significance between groups (* = 0.05<<i>P</i><0.01, ** = 0.01<<i>P</i><0.001, *** = 0.001<<i>P</i><0.0001, **** = <i>P</i><0.0001). WT, wild type; <i>Tph1</i><sup><i>-/-</i></sup>, <i>Tph1</i> deficient; LFD, low fat diet; HFD, high fat diet.</p

    Composition of Control and High Fat Diets<sup>a</sup>.

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    <p>Composition of Control and High Fat Diets<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162432#t001fn001" target="_blank"><sup>a</sup></a>.</p

    Elevating serotonin pre-partum alters the Holstein dairy cow hepatic adaptation to lactation

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    <div><p>Serotonin is known to regulate energy and calcium homeostasis in several mammalian species. The objective of this study was to determine if pre-partum infusions of 5-hydroxytryptophan (5-HTP), the immediate precursor to serotonin synthesis, could modulate energy homeostasis at the level of the hepatocyte in post-partum Holstein and Jersey dairy cows. Twelve multiparous Holstein cows and twelve multiparous Jersey cows were intravenously infused daily for approximately 7 d pre-partum with either saline or 1 mg/kg bodyweight of 5-HTP. Blood was collected for 14 d post-partum and on d30 post-partum. Liver biopsies were taken on d1 and d7 post-partum. There were no changes in the circulating concentrations of glucose, insulin, glucagon, non-esterified fatty acids, or urea nitrogen in response to treatment, although there were decreased beta-hydroxybutyrate concentrations with 5-HTP treatment around d6 to d10 post-partum, particularly in Jersey cows. Cows infused with 5-HTP had increased hepatic serotonin content and increased mRNA expression of the serotonin 2B receptor on d1 and d7 post-partum. Minimal changes were seen in the hepatic mRNA expression of various gluconeogenic enzymes. There were no changes in the mRNA expression profile of cell-cycle progression marker cyclin-dependent kinase 4 or apoptotic marker caspase 3, although proliferating cell nuclear antigen expression tended to be increased in Holstein cows infused with 5-HTP on d1 post-partum. Immunofluorescence assays showed an increased number of CASP3- and Ki67-positive cells in Holstein cows infused with 5-HTP on d1 post-partum. Given the elevated hepatic serotonin content and increased mRNA abundance of <i>5HTR2B</i>, 5-HTP infusions may be stimulating an autocrine-paracrine adaptation to lactation in the Holstein cow liver.</p></div

    Hepatic serotonin content and <i>TPH1</i> and <i>SERT</i> mRNA abundance of multiparous Holstein cows and multiparous Jersey cows administered pre-partum daily I.V. infusions of 1 liter of saline or 1 liter of 1.0 mg/kg bodyweight of 5-hyroxy-L-tryptophan (5-HTP) reconstituted in saline.

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    <p>Final treatment groups were saline-infused Holsteins (<i>n</i> = 6), 5-HTP infused Holsteins (<i>n</i> = 6), saline-infused Jerseys (<i>n</i> = 6), and 5-HTP infused Jerseys (<i>n</i> = 6). (A) Liver serotonin content corrected to ug of protein (B) mRNA abundance of tryptophan hydroxylase 1 (<i>TPH1</i>) and (C) mRNA abundance of serotonin reuptake transporter (<i>SERT</i>). Stars indicate statistical difference between group means (* = 0.05<<i>P</i><0.01). All values are reported as LS means ± SEM.</p

    Immunofluorescent images and counts of cells positive for Ki67 and CASP3 in multiparous Holstein cows and multiparous Jersey cows administered pre-partum daily I.V. infusions of 1 liter of saline or 1 liter of 1.0 mg/kg bodyweight of 5-hyroxy-L-tryptophan (5-HTP) reconstituted in saline.

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    <p>Final treatment groups were saline-infused Holsteins (<i>n</i> = 6), 5-HTP infused Holsteins (<i>n</i> = 6), saline-infused Jerseys (<i>n</i> = 6), and 5-HTP infused Jerseys (<i>n</i> = 6). (A) Percent of cells positive for Ki67 stain of all counted cells in representative images (<i>n</i> = 3) from each treatment group. (B) Immunofluorescent images of Ki67-positive cells. The scale bar indicates 15 microns. The arrow indicates an example of a cell considered positive for Ki67. d1 and d7 are days 1 and 7 post-partum, respectively. CON and 5-HTP represent saline-infused and 5-HTP infused groups, respectively. (C) Immunofluorescent images of CASP3-positive cells. The scale bar indicates 15 microns. The arrow indicates an example of a cell considered positive for CASP3. (D) Count of cells positive for CASP3 in representative images (<i>n</i> = 3) from each treatment group. Stars indicate statistical difference between group means (* = 0.05<<i>P</i><0.01; *** = 0.001<<i>P</i><0.0001). All values are reported as LS means ± SEM.</p
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