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Gene expression in a pure population of odontoblasts isolated by laser-capture microdissection.
J Dent Res 2001; 80(11):1963-7JD

Abstract

Studies of odontoblast differentiation and function have been limited due to difficulties in obtaining sufficient numbers of intact cells. We describe a novel approach of laser-capture microdissection to obtain homogenous populations of pre-odontoblasts and odontoblasts from tissue sections of mouse molar cusp tips. Fixation, processing, and staining conditions were assessed for the optimal retrieval of total RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA from three-day-old captured odontoblasts was sufficient in quantity and quality. Odontoblast-specific gene expression was studied by RT-PCR analysis performed in a single streptavidin-coated tube. At E15.5, Days 0 and 3, gene expression in laser-captured odontoblasts resembled that seen in vivo by in situ hybridization. The use of LCM is thus a valuable means of retrieving quality RNA from discrete populations of odontoblasts at different stages of dentinogenesis.

Authors+Show Affiliations

Department of Orthodontics, University of Texas at Houston Health Science Center, Dental Branch, 77030, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11759003

Citation

Hoffmann, M, et al. "Gene Expression in a Pure Population of Odontoblasts Isolated By Laser-capture Microdissection." Journal of Dental Research, vol. 80, no. 11, 2001, pp. 1963-7.
Hoffmann M, Olson K, Cavender A, et al. Gene expression in a pure population of odontoblasts isolated by laser-capture microdissection. J Dent Res. 2001;80(11):1963-7.
Hoffmann, M., Olson, K., Cavender, A., Pasqualini, R., Gaikwad, J., & D'Souza, R. N. (2001). Gene expression in a pure population of odontoblasts isolated by laser-capture microdissection. Journal of Dental Research, 80(11), pp. 1963-7.
Hoffmann M, et al. Gene Expression in a Pure Population of Odontoblasts Isolated By Laser-capture Microdissection. J Dent Res. 2001;80(11):1963-7. PubMed PMID: 11759003.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Gene expression in a pure population of odontoblasts isolated by laser-capture microdissection. AU - Hoffmann,M, AU - Olson,K, AU - Cavender,A, AU - Pasqualini,R, AU - Gaikwad,J, AU - D'Souza,R N, PY - 2002/1/5/pubmed PY - 2002/1/5/medline PY - 2002/1/5/entrez SP - 1963 EP - 7 JF - Journal of dental research JO - J. Dent. Res. VL - 80 IS - 11 N2 - Studies of odontoblast differentiation and function have been limited due to difficulties in obtaining sufficient numbers of intact cells. We describe a novel approach of laser-capture microdissection to obtain homogenous populations of pre-odontoblasts and odontoblasts from tissue sections of mouse molar cusp tips. Fixation, processing, and staining conditions were assessed for the optimal retrieval of total RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA from three-day-old captured odontoblasts was sufficient in quantity and quality. Odontoblast-specific gene expression was studied by RT-PCR analysis performed in a single streptavidin-coated tube. At E15.5, Days 0 and 3, gene expression in laser-captured odontoblasts resembled that seen in vivo by in situ hybridization. The use of LCM is thus a valuable means of retrieving quality RNA from discrete populations of odontoblasts at different stages of dentinogenesis. SN - 0022-0345 UR - https://www.unboundmedicine.com/medline/citation/11759003/Gene_expression_in_a_pure_population_of_odontoblasts_isolated_by_laser_capture_microdissection_ L2 - http://journals.sagepub.com/doi/full/10.1177/00220345010800110301?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -