Abstract
The main purpose of this study was to express human myoglobin in mouse brain neurons and investigate the effects of this expression on metabolism and blood flow using phosphorous (31P) NMR spectroscopy and NMR perfusion imaging. Transgenic mice expressing brain myoglobin were created using a cDNA sequence for human myoglobin placed under the transcriptional control of either a human platelet-derived grown factor polypeptide B (PDGF-B) promoter sequence or a rat neuron-specific enolase (NSE) promoter sequence. The presence of myoglobin having a functional, reduced-state, heme group was demonstrated by protein analysis and immunocytochemistry. Expression levels were highest in the hippocampus, cerebellum, and cerebral cortex. No gross morphological adaptations of neural tissue resulting from the expression were observed and no statistically significant differences in the energetic state, as measured by 31P NMR, or baseline cortical perfusion, as measured by an NMR perfusion imaging technique, were found.
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Abbreviations
- NSE:
-
neuron-specific enolase
- PDGF:
-
platelet-derived growth factor
References
Rosenthal MR, LaManna JC, Jöbsis FF, Levasseur JE, Kontos HA, and Patterson JL. Effects of respiratory gases on cytochrome a in intact cerebral cortex: Is there a critical pO2? Brain Res 1976;108:143.
Detre JA, Williams DS, and Koretsky AP. Nuclear magnetic resonance determination of flow, lactate, and phosphate metabolites during amphetamine stimulation of the rat brain. NMR Biomed 1990;3:272.
Gayeski TEJ, Connett RI, and Honig CR. Minimum intracellular pO2 for maximum cytochrome turnover in red muscle in situ. Am J Physiol 1987;252:H906.
Jones DP. Intracellular diffusion gradients of O2 and ATP. Am J Physiol 1986;250:C663.
Wittenberg BA and Wittenberg JB. Transport of oxygen in muscle. Annu Rev Physiol 1989;51:857.
Garry DJ, Ordway GA, Lorenz JN, Radford NB, Chin ER, Grange RW, Bassel-Duby R, and Williams RS. Mice without myoglobin. Nature 1999;395:905.
Adra CN, Boer PH, and McBurney MW. Cloning and expression of the mouse pgk-1 gene and the nucleotide sequence of its promoter. Gene 1987;60:65.
Choi T, Huang M, Gorman C, and Jaenisch R. A genetic intron increases gene expression in transgenic mice. Mol Cell Biol 1991;11:3070.
Potten H, Jendraschak E, Hauck S, Amar LC, Avner P, and Müllhofer G. Molecular cloning and sequencing of a murine pgk-1 pseudogene family. Gene 1988;71:461.
Varadarajan, Szabo RA, and Boxer SG. Cloning, expression in Escherichia coli, and reconstitution of human myoglobin. Proc Natl Acad Sci USA 1985;82:5681.
Ratner L, Thielan B, and Collins T. Sequences of the 5’ portion of the human c-sis gene:Characterization of the transcriptional promoter and regulation of expression of the protein product by 5’ untranslated mRNA sequences. Nucleic Acids Res 1987;15:6017.
Forss-Petter S, Danielson PE, Catsicas S, Battenberg E, Price J, Nerenberg M, and Sutcliffe JG. Transgenic mice expressing ß-galactosidase in mature neurons under neuron-specific enolase promoter control. Neuron 1990;5:187.
Hogan B, Beddington R, Costantini F, and Lacy E. Manipulating the Mouse Embryo: A Laboratory Manual. Plainview, NY: Cold Spring Harbor Laboratory Press, 1994.
Thomas PE, Ryan D, and Levin W. An improved staining procedure for the detection of the peroxidase activity of cytochrome P-450 on sodium dodecyl sulfate polyacrylamide gels. Anal Biochem 1976;75:168.
Silva AC, Zhang W, Williams DS, and Koretsky AP. Multi-slice MRI of rat brain perfusion during amphetamine stimulation using arterial spin labeling. Magn Reson Med 1995;33:209.
Williams DS, Detre JA, Leigh JS, and Koretsky AP. Magnetic resonance imaging of perfusion using spin inversion of arterial water. Proc Natl Acad Sci USA 1992;89:212.
Cole RP, Sukanek PC, Wittenberg JB, and Wittenberg BA. Mitochondrial function in the presence of myoglobin. J Appl Physiol. 1982;53:1116.
Jones DP and Kennedy F. Intracellular oxygen gradients in cardiac myocytes: Lack of a role for myoglobin in facilitation of intracellular oxygen diffusion. Biochem Biophys Res Commun. 1982;105:419.
Roy CW and Sherrington CS. On the regulation of the blood supply of the brain. J Physiol Lond 1890;11:85.
Fox PT and Raichle ME. Focal physiological uncoupling of cerebral blood flow and oxidative metabolism during somatosensory stimulation in human subjects. Proc Natl Acad Sci USA 1986;83:1140.
Arakaki LSL, Kushmerick MJ, and Bums DH. Myoglobin oxygen saturation measured independently of hemoglobin in scattering media by optical reflectance spectroscopy. Appl Spectrosc 1996;50:697.
Tamura M, Oshino N, Chance B, and Silver IA. Optical measurements of intracellular oxygen concentration of rat heart in vitro. Arch Biochem Biophys 1978 191:8.
Jue T and Anderson S. 1H NMR observation of tissue myoglobin: An indicator of cellular oxygenation in vivo. Magn Reson Med 1990;13:524.
Wang Z, Noyszewski EA, and Leigh JS Jr. In vivo MRS measurement of deoxymyoglobin in human forearms. Magn Reson Med 1990 14:562.
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Shonat, R.D., Koretsky, A.P. (2003). Expression of Myoglobin in the Transgenic Mouse Brain. In: Dunn, J.F., Swartz, H.M. (eds) Oxygen Transport to Tissue XXIV. Advances in Experimental Medicine and Biology, vol 530. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-0075-9_31
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DOI: https://doi.org/10.1007/978-1-4615-0075-9_31
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