PTH/PTHrP receptor in early development and Indian hedgehog-regulated bone growth

Science

Volumn 273, Issue 5275, 1996, Pages 663-666

  Lanske, Beate ^a   Karaplis, Andrew C ^b   Lee, Kaechong ^a   Luz, Arne ^c   Vortkamp, Andrea ^d   Pirro, Alison ^a   Karperien, Marcel ^e   Defize, Libert H K ^e   Ho, Chrystal ^f   Mulligan, Richard C ^g,h   Abou Samra, Abdul Badi ^a   Jüppner, Harald ^a   Segre, Gino V ^a   Kronenberg, Henry M ^a  

^a MASSACHUSETTS GENERAL HOSPITAL   (United States)

^b MCGILL UNIVERSITY   (Canada)

^c INSTITUTE OF EPIDEMIOLOGY   (Germany)

^d HARVARD MEDICAL SCHOOL   (United States)

^e HUBRECHT INSTITUTE   (Netherlands)

^f HOWARD HUGHES MEDICAL INSTITUTE   (United States)

^g WHITEHEAD INSTITUTE FOR BIOMEDICAL RESEARCH   (United States)

^h MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PARATHYROID HORMONE; PARATHYROID HORMONE RECEPTOR; PARATHYROID HORMONE RELATED PROTEIN;

ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; BONE DEVELOPMENT; BONE GROWTH; CARTILAGE CELL; CELL DIFFERENTIATION; GENE DELETION; GENE TARGETING; HEDGEHOG; LIGAND BINDING; NEGATIVE FEEDBACK; NONHUMAN; PRIORITY JOURNAL; REVIEW;

ANIMALIA; ERINACEIDAE; MURINAE;

EID: 9344241375     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5275.663     Document Type: Review

References (29)

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18. 7 plaque-forming units per milliliter) with the coding sequence of the rat PTH/PTHrP receptor as a probe (4, 27). For construction of the targeting vector, a 4.5-kb Bam HI fragment containing exon E1 of the PTH/PTHrP receptor gene was used as the 5′ flanking region and subcloned by blunt end ligation into a similarly treated Xho I site in the pPNT plasmid (14). A 5.6-kb Eco RI fragment containing the 3′ untranslated region of the PTH/PTHrP receptor gene, cloned into the Eco RI polylinker site of pPNT, was used as the 3′ flanking region. The resulting targeting vector was linearized at the unique Not I site in the pPNT backbone for electroporation of ES cells. Double selection was carried out, and resistant ES clones were analyzed by Southern (DNA) blot analyses with an external intronic 0.9-kb Sac I-Xho I fragment at the 5′ end of the gene. Positive clones (12%) were injected into recipient blastocysts to generate chimeras as described (28). Progeny of two independent clones yielded the identical phenotype. All animal experimentation followed institutional guidelines.
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29. 35S- labeled RNA probes were transcribed from the rat PTH/PTHrP receptor cDNA (4) and from the human lhh cDNA (16). In situ hybridization was done as described (8). 30. We thank C. Tabin for valuable collaboration and helpful reading of the manuscript, T. Doetschmann for reagents, E. Samson for technical assistance with histology, and M. Mannstadt for help and technical expertise. Supported by National Institutes of Health grants DK 47038 and DK 47237. B.L. was supported in part by a fellowship of the Max-Kade Foundation.