Functional development of fetal suprachiasmatic nucleus grafts in suprachiasmatic nucleus-lesioned rats.

Recovery of circadian drinking rhythms in suprachiasmatic nucleus (SCN)-lesioned rats after fetal SCN grafting was related to the immunocytochemical appearance and fiber outgrowth of vasopressin (VP)-, vasoactive intestinal polypeptide (VIP)-, and somatostatin (SOM)-containing neurons in the implants. At 4 weeks postgrafting, the first recovered animal was found. After longer survival times, 38% of the animals showed recovery.

Gelatin embedding to preserve lesion-damaged hypothalami and intracerebroventricular grafts for vibratome slicing and immunocytochemistry.

Vibratome sectioning of paraformaldehyde-fixed brains containing a lesion and/or intracerebroventricular grafts and staining of these sections often lead to damage around the site of the lesion and loss of the implants. Various embedding procedures were compared in order to find a method to overcome this problem. The best results were obtained when fixed brain tissue was first embedded under reduced pressure in 10% gelatin at 50 degrees C, and then fixed again, in 4% paraformaldehyde.

Grafted fetal suprachiasmatic nucleus cells survive much better in tissue pieces than in suspension.

A comparison was made between the survival of fetal suprachiasmatic nucleus (SCN) grafted either in tissue pieces or as tissue suspension. Donor tissue was obtained from day 15, 16 or 17 Wistar fetuses, and stereotaxically placed in the dorsal thalamus of the brain of vasopressin(VP)-deficient Brattleboro adult rats. One month post-grafting, the suspension grafts largely failed to show the immunocytochemical presence of VP- and vasoactive intestinal polypeptide(VIP)-producing SCN cells, but solid piece grafts did.

Developmental and functional aspects of grafting of the suprachiasmatic nucleus in the Brattleboro and the arrhythmic rat.

The development of suprachiasmatic nuclei (SCN) dissected from fetal rats and grafted in adult rat brains has provided additional insights in the normal ontogeny of the SCN. The SCN survives rather easily and develops to its typical adult cytoarchitectonical arrangement of contiguous clusters of vasopressin (VP)-, vasoactive intestinal polypeptide (VIP)- and somatostatin (SOM)- immunoreactive cells. Neither site of implantation, nor the establishment of efferent or afferent connections of the grafted SCN seems to be essential to allow it to develop normally into this distinguishing cytology.