Post-natal development of type 1 cannabinoid receptor immunoreactivity in the rat hippocampus.

Type 1 cannabinoid receptors, selectively located on axon terminals of GABAergic interneurons in the hippocampus, are known to be involved in endocannabinoid-mediated retrograde synaptic signalling. The question arises whether type 1 cannabinoid receptors appear on these axons during early post-natal life, when GABAergic transmission is still depolarizing, and whether there are any developmental changes in the cellular or subcellular expression pattern. Here we demonstrate, using single and double immunocytochemical methods at the light and electron microscopic levels, that type 1 cannabinoid receptors are expressed only on the membrane of axon terminals and pre-terminal axons but not on the soma-dendritic membrane at all examined timepoints between post-natal days 0 and 20, similar to the adult distribution. All type 1 cannabinoid receptor-positive boutons formed symmetric synapses. Granular labelling in the somata was already strong at post-natal day 0 and corresponded to multivesicular bodies, lysosomes, Golgi apparatus and rough endoplasmic reticulum. The type 1 cannabinoid receptor-positive axons were shown to originate largely from cholecystokinin-immunoreactive basket and bistratified neurons throughout the hippocampus (90% of all type 1 cannabinoid receptor-containing cells) and dentate gyrus (70% of all type 1 cannabinoid receptor-containing cells). The remaining cells have not been identified but probably belong to the somatostatin- and/or neuropeptide Y-containing subsets, as cholecystokinin-negative, type 1 cannabinoid receptor-positive axons have been observed in strata moleculare and lacunosum-moleculare of the dentate gyrus and CA1-3, respectively, where these neurons are known to arborize. No cell types were found that expressed type 1 cannabinoid receptors transiently at some developmental stage. We conclude that the cellular and subcellular pattern of type 1 cannabinoid receptor expression during early post-natal life is similar to the adult pattern and type 1 cannabinoid receptors are expressed on the cholecystokinin-containing axons as soon as synapse formation begins. This suggests that retrograde synaptic signalling by endocannabinoids is required for the normal operation of GABAergic neurotransmission even before it becomes hyperpolarizing.