Different regulatory sequences are required for parvalbumin gene expression in skeletal muscles and neuronal cells of transgenic mice.
Parvalbumin (PV) is expressed in fast-twitch fibers in skeletal muscles and a subpopulation of inhibitory neurons in the CNS. We generated transgenic mice that expressed the human interleukin-2 receptor alpha-subunit-green fluorescent fusion protein (hIL-2R-GFP) using two types of PV transgene. One contained the hIL-2R-GFP gene downstream of a 16.5-kb 5'-upstream PV genomic sequence (PV line). The other comprised the hIL-2R-GFP gene in bacterial artificial chromosome (BAC) with either a 180-kb (PA line) or 155-kb (PB line) insert encompassing the PV gene. Independent lines of all transgenic mice showed a faithful hIL-2R-GFP expression in fast-twitch muscle fibers. However, appreciable hIL-2R-GFP expression in the CNS occurred only in the PA transgenic lines. In one line of PA transgenic mice, hIL-2R-GFP was properly expressed in PV-containing neurons in the cerebellum, thalamic reticular nucleus, globus pallidus and cerebral cortex, though ectopic expression was observed in a particular subset of cerebellar astrocytes. Another line of PA transgenic mice showed a selective and mosaic expression of hIL-2R-GFP in PV-containing Purkinje, basket and stellate cells in the cerebellum. These results indicate that the 16.5-kb PV genomic sequence is sufficient for fiber-type-selective transcription but additional regulatory sequences comprised in BAC DNA are required for proper expression in PV-containing neurons.
Hazama
Masaaki
M
Department of Biological Sciences, Kyoto University Faculty of Medicine, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan.
Watanabe
Dai
D
Suzuki
Misao
M
Mizoguchi
Akira
A
Pastan
Ira
I
Nakanishi
Shigetada
S
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000502:physiology*; D005809:Genes, Regulator / Q000235:genetics*; D005822:Genetic Vectors / Q000502:physiology; D049452:Green Fluorescent Proteins; D000922:Immunotoxins / Q000494:pharmacology; D007202:Indicators and Reagents / Q000378:metabolism; D053645:Interleukin-2 Receptor alpha Subunit; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018656:Muscle Fibers, Fast-Twitch / Q000166:cytology / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D009433:Neural Inhibition / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010320:Parvalbumins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018123:Receptors, Interleukin / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D019076:Transgenes / Q000235:genetics; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
C508017:IL2RA protein, human; C508018:Il2ra protein, mouse; D000922:Immunotoxins; D007202:Indicators and Reagents; D053645:Interleukin-2 Receptor alpha Subunit; D008164:Luminescent Proteins; D010320:Parvalbumins; D012333:RNA, Messenger; D018123:Receptors, Interleukin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005680:gamma-Aminobutyric Acid