In cat four times as many lamina I neurons project to the parabrachial nuclei and twice as many to the periaqueductal gray as to the thalamus.

The spinothalamic tract, and especially its fibers originating in lamina I, is the best known pathway for transmission of nociceptive information. On the other hand, different studies have suggested that more lamina I cells project to the parabrachial nuclei (PBN) and periaqueductal gray (PAG) than to the thalamus. The exact ratio of the number of lamina I projections to PBN, PAG and thalamus is not known, because comprehensive studies examining these three projections from all spinal segments, using the same tracers and counting methods, do not exist. In the present study, the differences in number and distribution of retrogradely labeled lamina I cells in each segment of the cat spinal cord (C1-Coc2) were determined after large wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) injections in either PBN, PAG or thalamus. We estimate that approximately 6000 lamina I cells project to PBN, 3000 to PAG and less than 1500 to the thalamus. Of the lamina I cells projecting to thalamus or PAG more than 80%, and of the lamina I-PBN cells approximately 60%, were located on the contralateral side. In all cases, most labeled lamina I cells were found in the upper two cervical segments and in the cervical and lumbar enlargements.

How many spinothalamic tract cells are there? A retrograde tracing study in cat.

The spinothalamic tract, well known for its role in nociception, is the most frequently studied ascending pathway originating from the spinal cord. It is known that spinothalamic neurons are located in all segments of the spinal cord, but in most mammals the total number of spinothalamic neurons is not known. In three cats, after large wheat germ agglutinin-conjugated horseradish peroxidase injections involving all parts (one case) or almost all parts of the thalamus (two cases), the number of retrogradely labeled profiles was counted in a 1:4 series of sections of all spinal segments from C1 to Coc2. After applying the correction factor of Abercrombie (Anat. Rec. 94 (1946) 239), it appears that a total of 12,000 cells in the spinal cord project to the thalamus.

Less than 15% of the spinothalamic fibers originate from neurons in lamina I in cat.

Lamina I neurons sending their axons into the spinothalamic tract are thought to play a crucial role in nociception, but many spinothalamic fibers do not originate from lamina I neurons. In cat, no consensus exists about what percentage of the spinothalamic tract cells are located in lamina I. After wheat germ agglutinin-conjugated horseradish peroxidase injections that covered large parts of the thalamus, retrogradely labeled cells were plotted and counted in all segments of the spinal cord. Results show that, averaged over all spinal segments, the percentage of labeled lamina I neurons was 4.9-14.2%. These results demonstrate that, in contrast to what is concluded in several previous studies, lamina I in the cat provides only a limited part of the total spinal input to the thalamus.

Direct projections from the nucleus retroambiguus to cricothyroid motoneurons in the cat.

Vocalization can be elicited by stimulation in the periaqueductal gray (PAG). Light-microscopical tracing and physiological studies have revealed that the PAG uses the nucleus retroambiguus (NRA) as a relay to excite the vocalization muscle motoneurons. Direct NRA projections have been demonstrated to pharyngeal and abdominal wall muscle motoneurons, but not to laryngeal motoneurons. In two cats 0.1% cholera toxin subunit b was injected in the cricothyroid muscle of the larynx to retrogradely label its motoneurons, and 2.5% wheat germ agglutinin-horseradish peroxidase was injected into the NRA to anterogradely label its fibers. The electronmicroscopical results indicate that the NRA fibers make monosynaptic contacts with cricothyroid motoneuronal dendrites. Almost all NRA terminal profiles had asymmetrical synapses and contained mostly round or pleiomorphic vesicles, which strongly suggests that the NRA-cricothyroid motoneuronal projection is an excitatory pathway.

The parabrachial nucleus is a critical link in the transmission of short latency nociceptive information to midbrain dopaminergic neurons.

Many dopaminergic neurons exhibit a short-latency response to noxious stimuli, the source of which is unknown. Here we report that the nociceptive-recipient parabrachial nucleus appears to be a critical link in the transmission of pain related information to dopaminergic neurons. Injections of retrograde tracer into the substantia nigra pars compacta of the rat labelled neurons in both the lateral and medial parts of the parabrachial nucleus, and intra-parabrachial injections of anterograde tracers revealed robust projections to the pars compacta and ventral tegmental area. Axonal boutons were seen in close association with tyrosine hydroxylase-positive (presumed dopaminergic) and negative elements in these regions. Simultaneous extracellular recordings were made from parabrachial and dopaminergic neurons in the anaesthetized rat, during the application of noxious footshock. Parabrachial neurons exhibited a short-latency, short duration excitation to footshock while dopaminergic neurons exhibited a short-latency inhibition. Response latencies of dopaminergic neurons were reliably longer than those of parabrachial neurons. Intra-parabrachial injections of the local anaesthetic lidocaine or the GABA(A) receptor antagonist muscimol reduced tonic parabrachial activity and the amplitude (and in the case of lidocaine, duration) of the phasic response to footshock. Suppression of parabrachial activity with lidocaine reduced the baseline firing rate of dopaminergic neurons, while both lidocaine and muscimol reduced the amplitude of the phasic inhibitory response to footshock, in the case of lidocaine sometimes abolishing it altogether. Considered together, these results suggest that the parabrachial nucleus is an important source of short-latency nociceptive input to the dopaminergic neurons.

Direct projections from the sacral spinal cord to the medial preoptic area in cat and guinea pig.

The medial preoptic area (MPO) plays an important role in many behavioral, autonomic and endocrine functions, including micturition and genital responses. Although afferents of the MPO have been studied extensively, it is unknown whether direct lumbosacral-MPO projections exist that could convey afferent information from the pelvic organs. We hypothesized that, if such direct projections exist, MPO projecting cells would be located in the lateral part of the sacral cord, where primary afferents from pelvic and pudendal nerves terminate. We used retro- and anterograde tracing techniques in cat and guinea pig to study this. In cats, injections in the MPO resulted in labeled cells throughout the spinal cord, but with the highest density in the S1-S2 segments. In guinea pigs, labeled cells were found exclusively in the S1-S3 segments after MPO injections. Labeled cells in the sacral segments were not located in the lateral parts of the gray matter, but were found in the medial laminae VI-VII and dorsal lamina VIII in cats, and mainly in lamina X in guinea pigs. Anterograde tracing results after injections in the sacral cord in cats or guinea pigs showed labeled fibers in the MPO, just ventral to the anterior commissure. The central location of the cells of origin within the sacral cord, together with the termination pattern of the spino-MPO projections, strongly suggest a role for the spino-MPO pathway in the sensory relay of pelvic viscera, important for micturition and genital responses.