Molecular neuroscienceDistribution of cGMP-dependent protein kinase type I and its isoforms in the mouse brain and retina
Section snippets
Experimental animals
For experiments seven male and five female wild-type mice and two male and two female cGKI knockout mice (Wegener et al., 2002) were used. Animals were one- to three-month-old and had a 129/Sv, C57BL/6 or mixed 129Sv/C57BL6 genetic background. Results were independent of gender, age or genetic background of the analyzed mice. This study conformed to the German animal protection law and had been approved by the committee on animal care and welfare of the local government.
Western blot analysis
Mouse tissues were
Immunohistochemical detection of cGKI in the mouse CNS and retina
For immunohistochemistry, a rabbit polyclonal antiserum detecting both cGKI isoforms, cGKIα and cGKIβ, was used (cGKI common antibody). The specificity of the cGKI common antibody was validated by using recombinant proteins as well as tissues from wild-type and cGKI knockout mice (Wegener et al 2002, Feil et al 2003, Kleppisch et al 2003, Geiselhoringer et al 2004; see also below). Importantly, immunohistochemical staining of sections revealed signals in wild-type but not cGKI knockout tissues (
Discussion
Based on immunohistochemistry and immunoblot analysis with cGKI-specific antibodies, this study shows that the cGKI is more widely distributed in the CNS than reported previously. In addition, this is the first analysis of the distribution and relative expression level of the cGKIα and cGKIβ proteins in the brain. The specificity of the cGKI antibodies was demonstrated by immunostaining of tissues from cGKI knockout mice. This strategy is presumably superior to conventional controls for
Conclusion
This study suggests that the distribution and functional role of cGKI in the mammalian CNS is broader than previously thought. Together with the localization of NO synthase and cGMP (De Vente et al., 1998), the new expression data for cGKI should be useful to draw an anatomical map of the NO/cGMP/cGKI pathway in the brain and to correlate it with functional data.
Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft and the VolkswagenStiftung.
References (21)
- et al.
The dorsomedial hypothalamic nucleus and its role in ingestive behavior and body weight regulationlessons learned from lesioning studies
Physiol Behav
(2002) - et al.
Distribution of nitric oxide synthase and nitric oxide-receptive, cyclic GMP-producing structures in the rat brain
Neuroscience
(1998) - et al.
Localization of the cGMP-dependent protein kinases in relation to nitric oxide synthase in the brain
J Chem Neuroanat
(1999) - et al.
Distribution of IRAG and cGKI-isoforms in murine tissues
FEBS Lett
(2004) The biology of cyclic GMP-dependent protein kinases
J Biol Chem
(2005)- et al.
Protein kinase G I immunoreaction is colocalized with arginine-vasopressin immunoreaction in the rat suprachiasmatic nucleus
Neurosci Lett
(2002) - et al.
Identification of the amino acid sequences responsible for high affinity activation of cGMP kinase Ialpha
J Biol Chem
(1997) - et al.
Presynaptic role of cGMP-dependent protein kinase during long-lasting potentiation
J Neurosci
(2001) - et al.
Functional reconstitution of vascular smooth muscle cells with cGMP-dependent protein kinase I isoforms
Circ Res
(2002) - et al.
Impairment of LTD and cerebellar learning by Purkinje cell-specific ablation of cGMP-dependent protein kinase I
J Cell Biol
(2003)
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Present address: Interfakultäres Institut für Biochemie der Universität Tübingen, Hoppe-Seyler-Straβe 4, 72076 Tübingen, Germany.