The central pattern generator for locomotion in vertebrates is composed of different spinal neuronal populations that generate locomotor movement. In the lamprey spinal cord, several classes of interneurons have been identified based on morphologic and physiological criteria and integrated in the spinal cord circuits implicated in the generation of locomotion. However, the lack of histochemical markers for most of the interneurons makes it difficult to study whole populations along the spinal cord. We have investigated the immunoreactivity with antibodies raised against calbindin and calretinin. Several types of neurons could be classified: (1). strongly immunoreactive neurons located dorsomedially, (2). moderately immunoreactive neurons located laterally, (3). small weakly immunoreactive neurons, d). ventromedial neurons, (4). liquor contacting cells, and (5). motoneurons. The ventromedial group of calbindin-immunoreactive neurons also is immunoreactive for serotonin and, therefore, represents the ventromedial group of dopamine/serotonin spinal neurons. Some of the lateral calbindin-immunoreactive neurons may be CC-type cells (cells with caudal-crossed axons), because they are retrogradely labeled by tracer injections into the contralateral spinal cord. Other well-characterized cell types, such as sensory dorsal cells, lateral interneurons, descending propriospinal edge cells, and spinobulbar giant interneurons are negative for both calbindin and calretinin. Therefore, calbindin and calretinin are useful markers for the study of cell populations that may be integrated in locomotor circuits. Copyright 2002 Wiley-Liss, Inc.