Expression patterns of intestinal calcium transport factors and ex-vivo absorption of calcium in horses

Sprekeler, Nele; Müller, Tobias; Kowalewski, Mariusz P.; Liesegang, Annette; Boos, Alois

doi:10.1186/1746-6148-7-65

Cited by 15 publications

(7 citation statements)

References 33 publications

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“…Together these genes, TRPV5, TRPV6, calD 9k , calD 28k , PMCA, NCX1, CYP27B1, CYP24A1, and VDR regulate transcellular calcium transport in epithelial cells [ 3 , 11 ]. Different studies published on calcium related genes in mice [ 17 ], humans [ 18 , 19 ], sheep [ 20 – 23 ], rabbits [ 24 ], goats [ 21 – 23 ], horses [ 25 – 27 ], and dogs [ 28 , 29 ], demonstrated a correlation between the expression of calcium transport genes and the capacity of cells for calcium absorption [ 24 ]. Therefore, gene function is strongly related to their location and magnitude of expression.…”

Section: Discussionmentioning

confidence: 99%

Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney

et al. 2016

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The aim of this study was to determine the relative abundance and relationship of vitamin D responsive and calcium transporting transcripts (TRPV5, TRPV6, calD9k, calD28k, PMCA, NCX1, CYP27B1, CYP24A1, and VDR) in ovine, canine and, equine kidney using quantitative real-time PCR (RT-qPCR), and then perform a comparison between the three species. Renal tissue samples were harvested post-mortem from 10 horses, 10 sheep, and five dogs. Primers were designed for each gene. For each sample total RNA was extracted, cDNA synthesised, and RT-qPCR was performed. RT-qPCR data were normalised and statistical comparison was performed. Due to their consistent correlation with each other in each species, TRPV6, calD9k/calD28k, and PMCA appeared to be the main pathways involved in active transepithelial calcium transport in the kidney of sheep, dogs and horses. The results indicate that all of the studied genes were expressed in the renal tissue of studied species, although the expression levels and correlation of transcripts with each other were different from species to species. All vitamin D responsive and calcium transporting transcripts were highly correlated with VDR in equine kidney, but not in sheep and dogs. The CYP27B1 and CYP24A1 mRNAs showed a different renal expression pattern and correlation in horses compared with sheep and dogs. Given the high urinary calcium concentration and low serum 1,25(OH)2D concentration in horses, it could be expected that CYP27B1 expression would be lower than CYP24A1 in the horse, and this did not appear to be the case. The findings suggest that despite low serum vitamin D concentrations, vitamin D still plays a significant role in calcium metabolism in horses, especially given the strong correlations between VDR and vitamin D responsive transcripts in these animals.

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Section: Discussionmentioning

confidence: 99%

Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney

et al. 2016

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“…Real Time (Taqman) PCR was performed using the ABI 7500 Fast Real-Time PCR System (Applied Biosystems, Rotkreuz, Switzerland), as described previously [ 20 ]. In brief, samples were amplified in duplicates in a 25-μl reaction mixture containing 12.5 μl FastStart Universal Probe Master (Roche Diagnostics, Mannheim, Germany), 300 nM of each primer, 200 nM Taqman probe and 5 μl cDNA.…”

Section: Methodsmentioning

confidence: 99%

TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

Sprekeler

Kowalewski

Boos

2012

Reprod Biol Endocrinol

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BackgroundTransient receptor potential channel type 6 (TRPV6) and Calbindin-D9k (CaBP-9k) are involved in the active calcium (Ca2+) transport mechanism in many tissues including placenta and uterus, suggesting a role in the establishment and maintenance of pregnancy. Moreover, TRPV6 and CaBP-9k seem to support the materno-fetal Ca2+ transport that is crucial for fetal Ca2+ homeostasis, bone growth and development. However, it is unknown if these proteins are also involved in the aetiology of pathologies associated with parturition in cows, such as retained fetal membranes (RFM). The aim of the current study was to create an expression profile of uterine and placentomal TRPV6 and CaBP-9k mRNAs and proteins during pregnancy and postpartum in cows with and without fetal membrane release.MethodsUteri and placentomes of 27 cows in different stages of pregnancy and placentomes of cows with and without RFM were collected. Protein and mRNA expression of TRPV6 and CaBP-9k was investigated by real-time PCR, immunohistochemistry and Western blot.ResultsIn the uterine endometrium, highest TRPV6 and CaBP-9k expression was found in the last trimester of pregnancy, with a particular increase of protein in the glandular epithelium. In the placentomes, a gradual increase in TRPV6 mRNA was detectable towards parturition, while protein expression did not change significantly. Placentomal CaBP-9k expression did not change significantly throughout pregnancy but immunohistochemistry revealed an increase in staining intensity in the maternal crypt epithelium. Immunohistochemical, stronger placental CaBP-9k signals were seen in animals with RFM compared to animals with an undisturbed fetal membrane release, while protein levels, measured by Western blot analyses did not change significantly.ConclusionsThe results of the present study demonstrate a dynamic expression of TRPV6 and CaBP-9k during pregnancy in the bovine uterine endometrium and placentomes, suggesting a functional role for these proteins in Ca2+ metabolism during pregnancy. The temporal and spatial expression patterns indicate that TRPV6 and CaBP-9k may be involved in materno-fetal Ca2+ transport, mainly through an interplacentomal transport, and that both proteins may participate in physiological processes that are crucial for fetal and placental development. However, neither TRPV6 nor CaBP-9k seem to be causative in the retention of fetal membranes.

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“…Mucosal layers were mechanically removed, frozen in liquid nitrogen and stored until being analysed at À80°C. Protein preparation for VDR detection was carried out as described by Sprekeler et al (2011) with modifications. The antibody directed against VDR (Abcam AB54387; Sigma-Aldrich ChemieGmbH, Buchs, Switzerland) was diluted 1:200 in TBST containing 3% nonfat dry milk and 3% BSA.…”

Section: Western Blot Analysismentioning

confidence: 99%

Influence ofUVBexposure on the vitamin D status and calcium homoeostasis of growing sheep and goats

Kovács

Wilkens

Liesegang

2015

Animal Physiology Nutrition

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SummaryThe purpose of this study was to investigate the influence of exposure to ultraviolet radiation (UVB) on vitamin D status, intestinal calcium absorption and bone metabolism in growing sheep and goats. The hypothesis was that growing sheep and goats are able to synthesise vitamin D within their skin as a result of UVB exposure and that respective consequences for their vitamin D blood levels and the associated parameters can be shown. Fourteen 18-week-old lambs and goat kids were kept in an UVB-free environment and randomly assigned to two groups. One group was daily exposed to UVB (300 watt) for 12 weeks, and the other served as a control group. Except for the exposure to UVB, all animals were kept under the same conditions and fed according to their requirements. Before the start of the experiment and every second week, blood samples were taken. Also the left metatarsus of each animal was analysed by quantitative computer tomography to test for bone mineral status before the start, in week 7 and at the end of the experiment. After 12 weeks, the animals were slaughtered and samples were taken from skin, gastrointestinal tract and kidney for further analyses. In this study, exposure to UVB led to increased serum 1,25-dihydroxyvitamin D (1,25VitD) levels in goat kids, whereas in lambs, serum 25-hydroxyvitamin D (25VitD) levels were increased. In both species UVB-exposed animals showed lower 7-dehydrocholesterol (7DHC) values in skin than their respective control groups. These results indicate that growing goat kids and lambs are able to synthesise vitamin D in the skin when being exposed to UVB.

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Expression patterns of intestinal calcium transport factors and ex-vivo absorption of calcium in horses

Cited by 15 publications

References 33 publications

Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney

Evaluation and Comparison of Vitamin D Responsive Gene Expression in Ovine, Canine and Equine Kidney

TRPV6 and Calbindin-D9k-expression and localization in the bovine uterus and placenta during pregnancy

Influence ofUVBexposure on the vitamin D status and calcium homoeostasis of growing sheep and goats

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