The external granule layer of the developing chick cerebellum generates granule cells and cells of the isthmus and rostral hindbrain

The external granule layer (EGL) on the dorsal surface of the developing cerebellum consists of neural progenitors originating from the rostral rhombic lip (RRL). The RRL and the EGL were thought to give rise exclusively to the granule neurons of the cerebellum (Alder et al., 1996). To study the fate of individual RRL cells, we used a retroviral library to mark clones in the chick embryo at Hamberger-Hamilton stages 10-12. RRL clones comprised the EGL and cerebellar granule cells, as expected. Surprisingly, however, as many as 50% of the RRL clones also contained cells ventral to the cerebellum proper. Ventral derivatives were found in clones with a medial origin, as well as in those with a lateral origin along the RRL. Some of the ventral progeny appeared to be in the process of migration, whereas others appeared to be differentiating neurons in the isthmus and the rostral hindbrain region, including the locus coeruleus (LC) and pontine reticular formation. Furthermore, the Phox2a marker of LC precursors was detected in the EGL within the anterior aspect of the cerebellum. A stream of cells originating in the EGL and expressing Phox2a was observed to terminate ventrally in the LC. These data demonstrate that single RRL progenitor cells are not restricted to producing only cerebellar granule cells; they produce both cerebellar granule cells and ventral derivatives, some of which become hindbrain neurons. They also suggest that some progeny of the EGL escape the cerebellum via the anterior aspect of the cerebellar peduncles, to contribute to the generation of ventral structures such as the LC.