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Neural expression of alpha-internexin promoter in vitro and in vivo.
J Cell Biochem. 2006 Feb 01; 97(2):275-87.JC

Abstract

alpha-Internexin is a 66 kDa neuronal intermediate filament protein found most abundantly in the neurons of the nervous systems during early development. To characterize the function of mouse alpha-internexin promoter, we designed two different expression constructs driven by 0.7 kb or 1.3 kb of mouse alpha-internexin 5'-flanking sequences; one was the enhanced green fluorescent protein (EGFP) reporter for monitoring specific expression in vitro, and the other was the cre for studying the functional DNA recombinase in transgenic mice. After introducing DNA constructs into non-neuronal 3T3 fibroblasts and a neuronal Neuro2A cell line by lipofectamine transfection, we observed that the expression of EGFP with 1.3 kb mouse alpha-internexin promoter was in a neuron-dominant manner. To establish a tissue-specific pattern in the nervous system, we generated a transgenic mouse line expressing Cre DNA recombinase under the control of 1.3 kb alpha-Internexin promoter. The activity of the Cre recombinase at postnatal day 1 was examined by mating the cre transgenic mice to ROSA26 reporter (R26R) mice with knock-in Cre-mediated recombination. Analyses of postnatal day 1 (P1) newborns showed that beta-galactosidase activity was detected in the peripheral nervous system (PNS), such as cranial nerves innervating the tongue and the skin as well as spinal nerves to the body trunk. Furthermore, X-gal-labeled dorsal root ganglionic (DRG) neurons showed positive for alpha-Internexin in cell bodies but negative in their spinal nerves. The motor neurons in the spinal cord did not exhibit any beta-galactosidase activity. Therefore, the cre transgene driven by mouse alpha-internexin promoter, described here, provides a useful animal model to specifically manipulate genes in the developing nervous system.

Authors+Show Affiliations

Department of Anatomy and Cell Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

16173078

Citation

Wang, Pei, et al. "Neural Expression of Alpha-internexin Promoter in Vitro and in Vivo." Journal of Cellular Biochemistry, vol. 97, no. 2, 2006, pp. 275-87.
Wang P, Wang SM, Hsieh CJ, et al. Neural expression of alpha-internexin promoter in vitro and in vivo. J Cell Biochem. 2006;97(2):275-87.
Wang, P., Wang, S. M., Hsieh, C. J., & Chien, C. L. (2006). Neural expression of alpha-internexin promoter in vitro and in vivo. Journal of Cellular Biochemistry, 97(2), 275-87.
Wang P, et al. Neural Expression of Alpha-internexin Promoter in Vitro and in Vivo. J Cell Biochem. 2006 Feb 1;97(2):275-87. PubMed PMID: 16173078.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Neural expression of alpha-internexin promoter in vitro and in vivo. AU - Wang,Pei, AU - Wang,Seu-Mei, AU - Hsieh,Chia-Ju, AU - Chien,Chung-Liang, PY - 2005/9/21/pubmed PY - 2006/6/10/medline PY - 2005/9/21/entrez SP - 275 EP - 87 JF - Journal of cellular biochemistry JO - J Cell Biochem VL - 97 IS - 2 N2 - alpha-Internexin is a 66 kDa neuronal intermediate filament protein found most abundantly in the neurons of the nervous systems during early development. To characterize the function of mouse alpha-internexin promoter, we designed two different expression constructs driven by 0.7 kb or 1.3 kb of mouse alpha-internexin 5'-flanking sequences; one was the enhanced green fluorescent protein (EGFP) reporter for monitoring specific expression in vitro, and the other was the cre for studying the functional DNA recombinase in transgenic mice. After introducing DNA constructs into non-neuronal 3T3 fibroblasts and a neuronal Neuro2A cell line by lipofectamine transfection, we observed that the expression of EGFP with 1.3 kb mouse alpha-internexin promoter was in a neuron-dominant manner. To establish a tissue-specific pattern in the nervous system, we generated a transgenic mouse line expressing Cre DNA recombinase under the control of 1.3 kb alpha-Internexin promoter. The activity of the Cre recombinase at postnatal day 1 was examined by mating the cre transgenic mice to ROSA26 reporter (R26R) mice with knock-in Cre-mediated recombination. Analyses of postnatal day 1 (P1) newborns showed that beta-galactosidase activity was detected in the peripheral nervous system (PNS), such as cranial nerves innervating the tongue and the skin as well as spinal nerves to the body trunk. Furthermore, X-gal-labeled dorsal root ganglionic (DRG) neurons showed positive for alpha-Internexin in cell bodies but negative in their spinal nerves. The motor neurons in the spinal cord did not exhibit any beta-galactosidase activity. Therefore, the cre transgene driven by mouse alpha-internexin promoter, described here, provides a useful animal model to specifically manipulate genes in the developing nervous system. SN - 0730-2312 UR - https://www.unboundmedicine.com/medline/citation/16173078/Neural_expression_of_alpha_internexin_promoter_in_vitro_and_in_vivo_ DB - PRIME DP - Unbound Medicine ER -