title: Generation and characterization of Col10a1-mcherry reporter mice.
abstract: We report here on the generation of a new fluorescent protein reporter transgenic mouse line, Col10a1-mCherry, which can be used as a tool to study chondrocyte biology and pathology. Collagen, Type X, alpha 1 (Col10a1) is highly expressed in hypertrophic chondrocytes and commonly used as a gene marker for this cell population. The Col10a1-mCherry reporter line was generated using a bacterial recombination strategy with the mouse BAC clone RP23-192A7. To aid in the characterization of this animal model, we intercrossed Col10a1-mCherry mice with Collagen, Type II, alpha 1 (Col2a1) enhanced cyan fluorescent protein (ECFP) reporter mice and characterized the expression of both chondrocyte reporters during embryonic skeletal development from days E10.5 to E17.5. Additionally, at postnatal day 0, Col10a1-mCherry reporter expression was compared to endogenous Col10a1 mRNA expression in long bones and revealed that mCherry fluorescence extended past the Col10a1 expression domain. However, in situ hybridization for mCherry was consistent with the zone of Col10a1 mRNA expression, indicating that the persistent detection of mCherry fluorescence was a result of the long protein half life of mCherry in conjunction with a very rapid phase of skeletal growth and not due to aberrant transcriptional regulation. Taking advantage of the continued fluorescence of hypertrophic chondrocytes at the chondro-osseus junction, we intercrossed Col10a1-mCherry mice with two different Collagen, Type 1, alpha 1, (Col1a1) osteoblast reporter mice, pOBCol3.6-Topaz and pOBCol2.3-Emerald to investigate the possibility that hypertrophic chondrocytes transdifferentiate into osteoblasts. Evaluation of long bones at birth suggests that residual hypertrophic chondrocytes and osteoblasts in the trabecular zone exist as two completely distinct cell populations. genesis 49:410-418, 2011.
authors.affiliation: Department of Reconstructive Sciences, School of Dental Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030, USA. pmaye@neuron.uchc.edu
authors.forename: Peter
authors.initials: P
authors.lastname: Maye
authors.forename: Yu
authors.initials: Y
authors.lastname: Fu
authors.forename: David L
authors.initials: DL
authors.lastname: Butler
authors.forename: Kumar
authors.initials: K
authors.lastname: Chokalingam
authors.forename: Yaling
authors.initials: Y
authors.lastname: Liu
authors.forename: Jane
authors.initials: J
authors.lastname: Floret
authors.forename: Mary Louise
authors.initials: ML
authors.lastname: Stover
authors.forename: Richard
authors.initials: R
authors.lastname: Wenstrup
authors.forename: Xi
authors.initials: X
authors.lastname: Jiang
authors.forename: Cindi
authors.initials: C
authors.lastname: Gooch
authors.forename: David
authors.initials: D
authors.lastname: Rowe
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002356:Cartilage / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D024121:Collagen Type X / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000196:embryology / Q000378:metabolism; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017403:In Situ Hybridization; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D000097573:Red Fluorescent Protein
chemical_list: D024043:Collagen Type II; D024121:Collagen Type X; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C447413:Col2a1 protein, mouse; C485182:Cyan Fluorescent Protein