Cellular neuroscienceDistribution and neuropeptide coexistence of nucleobindin-2 mRNA/nesfatin-like immunoreactivity in the rat CNS
Section snippets
Animals
Male Sprague–Dawley rats (6–8 weeks; Scanlab, Stockholm, Sweden) were housed under 12-h dark/light conditions (lights on at 06:00 h and off at 18:00 h) in a temperature-controlled environment with free access to standard rat chow and tap water. All experiments had been approved by the local ethical committee (Stockholms Norra Djurförsöksetiska Nämnd) and the studies were carried out in accordance with the European Communities Council Directive of 24 November 1986 (86/609/EEC). All efforts were
Results
The distribution of NUCB2 mRNA was investigated by in situ hybridization (Fig. 1). The four oligonucleotide probes, complementary to non-overlapping segments of the NUCB2 cDNA, tested separately, yielded qualitatively similar in situ hybridization patterns, albeit with varying signal strength. The highest signal-to-noise ratio resulted from a combination of probes against base pairs 438–486 and 1402–1450; in the results described below and figures this combination was used. Furthermore, the
Discussion
Here we have studied the rat CNS distribution of NUCB2 mRNA and its newly described protein fragment nesfatin-1 which has been ascribed anorexigenic properties. We show a wide distribution in the hypothalamus and a restricted set of other brain regions, as well as its colocalization with several neuropeptides implicated in the control of energy metabolism. The brain distribution is summarized schematically in Fig. 10 (NUCB2/nesfatin cells indicated on plates from Paxinos et al., 2007). Our
Acknowledgments
The authors gratefully acknowledge the donors of antisera used in this study (see Experimental Procedures). We also thank Drs. Staffan Cullheim and Tomas Hökfelt and their respective laboratories for sharing expertise and equipment, and Dr. David Lyons for helpful comments on the manuscript. Emanuela Santini is gratefully acknowledged for expert assistance with Western blot experiments. Financial support for this study and publication was provided by the European Neuroscience Institutes
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