Cannabinoids and novelty investigation: influence of age and duration of exposure.

Administration of the synthetic cannabinoid receptor agonist WIN 55,212-2 has been shown to increase indices of noradrenergic activity. Neuroanatomical, neurochemical and behavioral studies have provided evidence supporting a marked impact of cannabinoids on the rat coeruleo-cortical pathway. As activity of this pathway is implicated in setting specific attentional modes, the present study assessed the influence of acute and repeated systemic administration of WIN 55,212-2 on novelty investigation in adolescent and adult male rats by using the hole board behavioral paradigm. Animals were individually acclimated to the hole board for 10-min sessions over 3 days, and novel objects were introduced on the fourth day. Novelty-seeking behavior was measured by comparison of the average number of return visits to a hole containing a novel object versus the average number of return visits to an empty hole. While attenuation of novelty preference was observed in adult rats acutely treated with WIN 55,212-2, both acutely treated adolescent groups retained their preference for novelty. All groups treated with repeated administration of either drug or vehicle demonstrated novelty preference, and no differences were found in the measure of novelty investigation between the groups. Furthermore, this study reproduced findings showing significant differences in locomotor activity that did not coincide with differences in novelty-seeking behavior. These data thus suggest a complex effect of CB1 receptor modulation on novelty preference in the male rat that is modulated by age and treatment.

Agonist-induced internalization of corticotropin-releasing factor receptors in noradrenergic neurons of the rat locus coeruleus.

Corticotropin-releasing factor (CRF) acts within the locus coeruleus (LC), to modulate activity of the LC-norepinephrine (NE) system. Combining molecular and cellular approaches, we demonstrate CRF receptor (CRFr) mRNA expression in Sprague-Dawley rat LC and provide the first in vivo evidence for agonist-induced internalization of CRFr. CRFr mRNA was detected in LC micropunches by RT-PCR. In dual labelling immunofluorescence studies, tyrosine hydroxylase (TH) containing neurons exhibited CRFr labelling. At the ultrastructural level, immunogold-silver labelling for CRFr was localized to the plasma membrane of TH-immunoperoxidase labelled dendrites. CRF (100 ng) injection into the LC produced a robust neuronal activation that peaked 10-15 min after injection and was maintained for the duration of the recording. This was associated with CRFr internalization in LC neurons that was apparent at 5 and 30 min after injection. By 5 min after injection the ratio of cytoplasmic to total dendritic CRFr-labelling was 0.81 +/- 0.01 in rats injected with CRF and 0.59 +/- 0.02 in rats injected with artificial cerebrospinal fluid (ACSF; P < 0.0001). Enhanced internalization of CRFr was maintained at 30 min after CRF injection, with the ratio being 0.86 +/- 0.02 for CRF-injected cases and 0.57 +/- 0.03 for ACSF-injected cases (P < 0.0001). Internalized CRFr was associated with early endosomes, indicative of degradation or recycling. Agonist-induced CRFr internalization in LC neurons may underlie acute desensitization to CRF or stress. This process may be a pivotal target by which stressors or pharmacological agents regulate the sensitivity of the LC-NE system to CRF and subsequent stressors.