Production and analysis of transgenic mice with ectopic expression of parvalbumin.
Transgenic mice expressing rat parvalbumin under the control of the human metallothionein IIA (MTII A), SV-40 early, and neuron-specific enolase (NSE) promoters were produced. Ectopic expression was analyzed by RNA polymerase chain reaction and RNase protection in combination with immunohistochemistry. From a total of 25 transgenic lines 18 were found to express the transgene. Expression strength and tissue specificity were dependent upon the promoter used and varied considerably among animal lines produced with the same construct. Highest constitutive MT IIA-driven expression was found in lung, liver, heart, and kidney, as well as in brain, and lower amounts of transgene expression were found in spleen, testis, and muscle. Immunohistochemistry of tissue sections of metallothionein-parvalbumin transgenic strain 29 in the non-induced state revealed that ectopic PV mRNA is translated into protein. Short-term induction of the MT IIA promoter by CdSO4 or CdCl2 leads to a shift in tissue specificity and does not increase ectopic expression in tissues where the transgene is active in the noninduced state. As expected the NSE promoter showed highest activity in brain. However, NSE-driven expression could also be detected to various degrees in all investigated tissues. SV-40-dependent PV expression showed no tissue preference and varied considerably among different strains. Except for the observation that the SV-40-PV construct showed lower yields in transgenic production and reduced numbers of positive offspring no obvious impairment of growth or behavior as a consequence of transgenic PV expression could be detected.
Castillo
M B
MB
Institut d'Histologie, Université de Fribourg Pérolles, Switzerland.
Celio
M R
MR
Andressen
C
C
Gotzos
V
V
Rülicke
T
T
Berger
M C
MC
Weber
J
J
Berchtold
M W
MW
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D018076:DNA, Complementary; D018014:Gene Transfer Techniques; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic / Q000378:metabolism*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010320:Parvalbumins / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012313:RNA / Q000032:analysis
D018076:DNA, Complementary; D010320:Parvalbumins; D012313:RNA