Generation patterns of immunocytochemically identified cholinergic neurons in rat brainstem

Combined [3H]thymidine autoradiographic and choline acetyltransferase (ChAT)-immunocytochemical techniques were used to answer questions concerning the generation of specific classes and subclasses of cholinergic neurons in rat brainstem. First, the generation of rostrally and caudally located neurons of the same class (i.e. somatic efferent oculomotor and hypoglossal nuclei, respectively) were compared. Results indicated that, although embryonic day 11 (E11) was the peak birthday for both nuclei, hypoglossal neurons were generated significantly earlier than oculomotor neurons, indicating a caudorostral generation gradient for brainstem somatic motor nuclei. Second, the generation patterns of 3 different subclasses of motor neurons at the same brainstem level were compared; namely those of the somatic efferent hypoglossal nucleus (XII), the general visceral efferent dorsal nucleus of the vagus (X), and the predominantly special visceral efferent nucleus ambiguus. All 3 subclasses of cholinergic cells had the same peak day (E11) and overall period of generation (E11-12). However, statistical analyses indicated a precocious generation of nucleus ambiguus, but no developmental differences between N, XII and N. X. It is suggested that nucleus ambiguus is formed earlier than N. XII and N. X, due to its more ventral location within a ventrodorsal neurogenetic gradient. Third, the generation patterns of different classes of large cholinergic neurons were examined. Specifically, the birthdays of cholinergic non-motor projection neurons of the pedunculopontine-laterodorsal tegmental nuclei (PPT-LDT) were contrasted to those of the cholinergic brainstem motor neurons. The peak birthdays of both rostrally and caudally located motor neurons were two days earlier than those of the PPT-LDT neurons. Thus, large cholinergic cells projecting to peripheral targets are born significantly earlier than those projecting within the CNS, even though the former are located more rostrally on the caudorostral neurogenetic gradient. This represents an apparent exception to the emerging rule that cholinergic neurons obey the general gradients of neurogenesis manifest in the regions of the central nervous system where they reside.