Quantitative and neurogenic analysis of the total population and subpopulations of neurons defined by axon projection in the superficial dorsal horn of the rat lumbar spinal cord

Abstract

The total neuron population of the superficial dorsal horn (SDH), i.e., laminae I and II, was quantitated in Nissl preparations of spinal segment L1 in the rat. Subpopulations of the SDH, defined by axon projection, were quantitated following strategic intraspinal injections of dual retrograde tracers (Fluoro-Gold and true blue). These methods were used in conjunction with [3H]thymidine (delivered in utero) autoradiography for neurogenic pattern analysis. Following stereological correction, each dorsal horn in spinal segment L1 contained 11 neurons in lamina I and 42.6 neurons in lamina II per 10-microm transverse section. Neurons with long projections, i.e., neurons with projections rostral to spinal segment T5, were only slightly more numerous in lamina I than in lamina II. These neurons made up 34% of the total neuron population in lamina I and 7.0% in lamina II. Most of these neurons did not demonstrate descending connections, and many (presumed supraspinal projection neurons) did not demonstrate short, ascending, intersegmental connections. Neurons with short propriospinal projections, i.e., neurons with connections caudal to spinal segment T5, made up approximately half of the total neuron population in both lamina I and lamina II: 55% and 52%, respectively. Of these, 79% had both short ascending and descending projections; the remaining 21% had only descending projections. Neurons that were not labeled with retrograde tracers (presumed local circuit cells) represented 11% of the neurons in lamina I and 41% in lamina II. Neurogenesis in the SDH proceeded along an axon-length gradient, whereby neurons with the longest axons completed neurogenesis first, and those with the shortest completed neurogenesis last. The generation of both propriospinal and supraspinal projection neurons began on embryonic day 13 (E13). Nearly equal numbers of neurons in this group were generated in laminae I and II through E14. On E15, neuron production slowed in lamina I and accelerated in lamina II as local circuit neurons and the remaining propriospinal neurons were generated. Neuron production ceased simultaneously in both lamina I and lamina II on E16.