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Transplantation of hypocretin neurons into the pontine reticular formation: preliminary results.

Arias-Carrión, Oscar; Murillo-Rodriguez, Eric; Xu, Man; Blanco-Centurion, Carlos; Drucker-Colín, Rene; Shiromani, Priyattam J.
Sleep ; 27(8): 1465-70, 2004 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-15683135


The sleep disorder narcolepsy is now considered a neurodegenerative disease because there is a massive loss of neurons containing the neuropeptide, hypocretin, and because narcoleptic patients have very low cerebrospinal fluid levels of hypocretin. Transplants of various cell types have been used to induce recovery in a variety of neurodegenerative animal models. In models such as Parkinson disease, cell survival has been shown to be small but satisfactory. Currently, there are no data indicating whether hypocretin neurons can survive when grafted into host tissue. Here we examined the survival of hypocretin-containing neurons grafted into the pontine reticular formation, a region traditionally regarded to be key for rapid eye movement sleep generation.


In 2 experiments, a suspension of cells from the posterior hypothalamus of 8- to 10-day old rat pups was injected into the pons (midline, at the level of the locus coeruleus) of adult rats. Control rats received cells from the cerebellum, tissue that is devoid of hypocretin neurons. In the first experiment (n = 33), the adult rats were sacrificed 1, 3, 6, 12, 24, or 36 days after transplant, and cryostat-cut coronal sections of the brainstem were examined for presence of hypocretin-immunoreactive neurons. In the second experiment (n = 9), the transplant medium was modified to include agents that stimulate cell growth, and recipient rats were sacrificed 9, 12, and 36 days after receiving the graft. SETTINGS Basic neuroscience research laboratory. MEASUREMENTS AND


In the first experiment, clearly defined hypocretin-immunoreactive containing somata and varicosities were visible in pons of rats sacrificed 1 day after grafting of posterior hypothalamic cells but not in rats receiving cerebellum tissue. The hypocretin-immunoreactive somata were not visible in rats sacrificed at 12, 24, or 36 days, indicating that the neurons had died. However, in the second experiment, where enriched transplant medium was used, clearly defined hypocretin-immunoreactive somata with processes and varicosities were present in the graft zone 36 days after implant. These somata were similar in size and appearance to adult rat hypocretin-immunoreactive neurons.


These results indicate that hypocretin neurons obtained from rat pups can be grafted into a host brain, and efforts should be made to increase survival of these neurons.