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Transgenic mouse model for conditional expression of influenza hemagglutinin-tagged human SLC20A1/PIT1.
PLoS One 2019; 14(10):e0223052Plos

Abstract

To further investigate the role of the phosphate (Pi) transporter PIT1 in Pi homeostasis and tissue mineralization, we developed a transgenic mouse expressing the C-terminal influenza hemagglutinin (HA) epitope-tagged human PIT1 transporter under control of the cytomegalovirus/chicken beta actin/rabbit beta-globin gene (CAG) promotor and a loxP-stop-loxP (LSL) cassette permitting conditional activation of transgene expression (LSL-HA-hPITtg/+). For an initial characterization of this conditional mouse model, germline excision of the LSL cassette was performed to induce expression of the transgene in all mouse tissues (HA-hPIT1tg/+). Recombination was confirmed using genomic DNA obtained from blood samples of these mice. Furthermore, expression of HA-hPIT1 was found to be at least 10-fold above endogenous mouse Pit1 in total RNA isolated from multiple tissues and from cultured primary calvaria osteoblasts (PCOB) estimated by semi-quantitative RT-PCR. Robust expression of the HA-hPIT1 protein was also observed upon immunoblot analysis in most tissues and permits HA-mediated immunoprecipitation of the transporter. Characterization of the phenotype of HA-hPIT1tg/+ mice at 80 days of age when fed a standard chow (0.7% Pi and 1% calcium) showed elevated plasma Pi, but normal plasma iPTH, iFGF23, serum calcium, BUN, 1,25-dihydroxy vitamin D levels and urine Pi, calcium and protein excretion when compared to WT littermates. Likewise, no change in bone mineral density was observed upon uCT analysis of the distal femur obtained from these mice. In conclusion, heterozygous overexpression of HA-hPIT1 is compatible with life and causes hyperphosphatemia while bone and mineral metabolism of these mice are otherwise normal.

Authors+Show Affiliations

Section of Endocrinology and Metabolism, Yale University School of Medicine, New Haven, CT, United States of America.Section of Endocrinology and Metabolism, Yale University School of Medicine, New Haven, CT, United States of America.Section of Endocrinology and Metabolism, Yale University School of Medicine, New Haven, CT, United States of America.Section of Endocrinology and Metabolism, Yale University School of Medicine, New Haven, CT, United States of America.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31613887

Citation

Chande, Sampada, et al. "Transgenic Mouse Model for Conditional Expression of Influenza Hemagglutinin-tagged Human SLC20A1/PIT1." PloS One, vol. 14, no. 10, 2019, pp. e0223052.
Chande S, Ho B, Fetene J, et al. Transgenic mouse model for conditional expression of influenza hemagglutinin-tagged human SLC20A1/PIT1. PLoS ONE. 2019;14(10):e0223052.
Chande, S., Ho, B., Fetene, J., & Bergwitz, C. (2019). Transgenic mouse model for conditional expression of influenza hemagglutinin-tagged human SLC20A1/PIT1. PloS One, 14(10), pp. e0223052. doi:10.1371/journal.pone.0223052.
Chande S, et al. Transgenic Mouse Model for Conditional Expression of Influenza Hemagglutinin-tagged Human SLC20A1/PIT1. PLoS ONE. 2019;14(10):e0223052. PubMed PMID: 31613887.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transgenic mouse model for conditional expression of influenza hemagglutinin-tagged human SLC20A1/PIT1. AU - Chande,Sampada, AU - Ho,Bryan, AU - Fetene,Jonathan, AU - Bergwitz,Clemens, Y1 - 2019/10/15/ PY - 2019/07/07/received PY - 2019/09/12/accepted PY - 2019/10/16/entrez PY - 2019/10/16/pubmed PY - 2019/10/16/medline SP - e0223052 EP - e0223052 JF - PloS one JO - PLoS ONE VL - 14 IS - 10 N2 - To further investigate the role of the phosphate (Pi) transporter PIT1 in Pi homeostasis and tissue mineralization, we developed a transgenic mouse expressing the C-terminal influenza hemagglutinin (HA) epitope-tagged human PIT1 transporter under control of the cytomegalovirus/chicken beta actin/rabbit beta-globin gene (CAG) promotor and a loxP-stop-loxP (LSL) cassette permitting conditional activation of transgene expression (LSL-HA-hPITtg/+). For an initial characterization of this conditional mouse model, germline excision of the LSL cassette was performed to induce expression of the transgene in all mouse tissues (HA-hPIT1tg/+). Recombination was confirmed using genomic DNA obtained from blood samples of these mice. Furthermore, expression of HA-hPIT1 was found to be at least 10-fold above endogenous mouse Pit1 in total RNA isolated from multiple tissues and from cultured primary calvaria osteoblasts (PCOB) estimated by semi-quantitative RT-PCR. Robust expression of the HA-hPIT1 protein was also observed upon immunoblot analysis in most tissues and permits HA-mediated immunoprecipitation of the transporter. Characterization of the phenotype of HA-hPIT1tg/+ mice at 80 days of age when fed a standard chow (0.7% Pi and 1% calcium) showed elevated plasma Pi, but normal plasma iPTH, iFGF23, serum calcium, BUN, 1,25-dihydroxy vitamin D levels and urine Pi, calcium and protein excretion when compared to WT littermates. Likewise, no change in bone mineral density was observed upon uCT analysis of the distal femur obtained from these mice. In conclusion, heterozygous overexpression of HA-hPIT1 is compatible with life and causes hyperphosphatemia while bone and mineral metabolism of these mice are otherwise normal. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/31613887/Transgenic_mouse_model_for_conditional_expression_of_influenza_hemagglutinin-tagged_human_SLC20A1/PIT1 L2 - http://dx.plos.org/10.1371/journal.pone.0223052 DB - PRIME DP - Unbound Medicine ER -