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. Author manuscript; available in PMC: 2012 Mar 7.
Published in final edited form as: J Neurosci. 2011 Sep 7;31(36):12708–12715. doi: 10.1523/JNEUROSCI.2172-11.2011

Figure 2. Axonal proteins in the pineal represent mainly sympathetic axonal arborsoriginating from the two superior cervical ganglia (SCG).

Figure 2

(A) Western blot analysis of βIII tubulin and TH in pineals from control and ganglionectomized animals (2xSCG) following 6, 12, and 36 hours after surgical denervation. Note the progressive decrease in TH (64% of control at 6h, 17% at 12h, and 1% at 36h 2xSCG) and βIII tubulin (53% of control at 6h, 40% at 12h, and 3% at 36h post-denervation) proteins following bilateral ganglionectomy. Expression of AANAT, which is induced by norepinephrine release from the SCG axon terminals, was monitored to confirm the surgical denervation. (B) Pineal glands from control (a–c) and 36h following 2xSCG, (d–f) were analyzed by immunohistochemistry for the expression of axonal markers βIII tubulin (a,d,c,f) and TH (b,e,c,f). In blue, Hoechst 3332 was used as nuclear counterstain. Note that,βIII tubulin and TH co-localize in axonal fibers in the control sample (a–c) and that, 36h following surgical denervation, axonal proteins are undetectable. All animals were sacrificed at ZT15. These results (panels A and B) were confirmed in more than 5 independent experiments.