Ann K. Goodchild

A method for evoking physiological responses by stimulation of cell bodies, but not axons of passage, within localized regions of the central nervous system

A method for evoking physiological responses by microinjection of sodium glutamate solution into localized regions of the central nervous system (CNS) is described. The major advantage of this method is that the cell bodies or dendritic processes of neurones within the injection site are excited, whereas axons of passage are unaffected. It was demonstrated that injections of minute volumes (50-100 nl) of 0.5 M glutamate solution into selected sites within the medulla or midbrain of anaesthetized or conscious animals, respectively, elicited marked autonomic, somatomotor or behavioural responses, depending on the injection site. In contrast, glutamate microinjection into fibre tracts failed to elicit any response, whereas electrical stimulation applied at the same sites elicited marked responses. The degree of localization of the glutamate stimulus and the relation between glutamate concentration and magnitude of evoked response are described. It is concluded that this method is a very effective means of selectively stimulating cell bodies within highly localized regions of the CNS. Further, by using this method in combination with focal electrical stimulation, it is possible in some cases to provide evidence that a response arises from excitation of axons of passage rather than cell bodies.

Role of ventrolateral medulla in vasomotor regulation: a correlative anatomical and physiological study

Two groups of experiments were carried out in rabbits. First, the ventrolateral reticular formation of the medulla oblongata was stimulated either by microinjection of sodium glutamate solution (exciting only cell bodies) or electrically (exciting cell bodies and axons). This region has been shown previously to contain a dense and compact group of bulbospinal cells. The effects of both electrical and chemical stimulation of specific sites were correlated with the density of ventrolateral bulbospinal cells at the same sites. Glutamate microinjection into the center of the group of bulbospinal cells elicited a very large and sustained increase in arterial pressure, whereas microinjection into sites outside this region elicited a very small or no response. These results suggest that it is the bulbospinal ventrolateral cells which mediate the pressor response to glutamate stimulation. Focal electrical stimulation in the ventrolateral medulla elicited increases in arterial pressure and decreases in femoral and renal vascular conductance, as well as a short-latency increase in renal sympathetic nerve activity. The most effective sites for focal electrical stimulation lay within the region of greatest density of bulbospinal cells; slightly less effective sites lay just rostral and caudal to this region. It is suggested that stimulation in these latter sites predominantly excites axons of passage. Secondly, the origin of afferent fibers to the ventrolateral vasomotor area was studied using the horseradish peroxidase (HRP) method. This revealed major projections from the medial part of the nucleus tractus solitarius and the parabrachial nucleus in the pons. The physiological and anatomical studies taken together are consistent with the hypothesis that the bulbospinal ventrolateral cells are vasomotor in function, and receive afferent inputs from brain stem nuclei which are known to play a role in autonomic regulation.

Evidence that Blue‐on Cells are Part of the Third Geniculocortical Pathway in Primates

Colour vision in primates is mediated by cone opponent ganglion cells in the retina, whose axons project to the dorsal lateral geniculate nucleus in the visual thalamus. It has long been assumed that cone opponent ganglion cells project to the parvocellular layers of the geniculate. Here, we examine the role of a third subdivision of the geniculocortical pathway: the interlaminar or koniocellular geniculate relay cells. We made extracellular recordings in the dorsal lateral geniculate nucleus of the common marmoset Callithrix jacchus, a New World monkey in which the interlaminar cells are well segregated from the parvocellular layers. We found that one group of colour opponent cells, the blue‐on cells, was largely segregated to the interlaminar zone. This segregation was common to dichromatic (‘red‐green colour‐blind’) and trichromatic marmosets. The result calls into question the traditional notion that all colour information passes through the parvocellular division of the retino‐geniculo‐cortical pathway in primates.

Cell groups in the lower brain stem of the rabbit projecting to the spinal cord, with special reference to catecholamine-containing neurons

Two groups of experiments were carried out in rabbits. In the first groups, the distribution of cell bodies within the pons and medulla projecting ipsilaterally and contralaterally to the thoracic or lumbar spinal cord was studied using the horseradish peroxidase (HRP)/tetramethylbenzidine (TMB) procedure. In the second group, both a previously described double-labeling technique and a new modification of it were used to determine the location of catecholamine (CA)-fluorescent pontomedullary cells projecting to the spinal cord. The results demonstrate that the catecholamine (probably norepinephrine)-containing neurons which innervate the thoracic spinal cord are confirmed almost exclusively to the pons where they were found within the A5, A7 and subcoeruleus groups, as well as the ventral portion of the principal part of the locus coeruleus and the more caudal locus coeruleus, including the A4 cell group. Within the medulla oblongata no doubly labeled A2 cells were observed and the few double labeled A1 cells which were observed were confined to the rostral portion of this group. A dense group of HRP-positive but non-fluorescent cells was found rostral to the A1 area in the ventrolateral reticular formation. These cells, which correspond in position to PNMT-containing cells in the rat, appear to project to both thoracic and lumbar segments of the spinal cord. In contrast, spinally projecting neurons within the nucleus tractus solitarius originated from different subnuclei according to their segmental destination. New information about the organization of medial reticulospinal and vestibulospinal pathways was also obtained.

Comparison of photoreceptor spatial density and ganglion cell morphology in the retina of human, macaque monkey, cat, and the marmoset Callithrix jacchus

We studied the relationship between the morphology of ganglion cells and the spatial density of photoreceptors in the retina of two Old World primates, human and macaque monkey; the diurnal New World marmoset Callithrix jacchus; and the cat. Ganglion cells in macaque and marmoset were labelled by intracellular injection with Neurobiotin or by Dil diffusion labelling in fixed tissue. Cone photoreceptor densities were measured from the same retinas. Supplemental data for macaque and data for human and cat were taken from published studies.

Retinal ganglion cells in the albino rat: Revised morphological classification

Rat retinal ganglion cells were traditionally classified on the basis of soma size and the morphology of their dendritic fields. However, in the past, techniques used to label ganglion cells (horseradish peroxidase, Golgi, or the neurofibrillar stain) did not always stain the axon and/or the entire dendritic field. In the present study, we have labelled retinal ganglion cells in the adult albino rat with the carbocyanine dye 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethylindo‐carbocyanine perchlorate (DiI) or have intracellularly injected them with Neurobiotin. Such procedures enabled us to completely fill these neurons, and our findings prompted us to modify the existing retinal ganglion cell classification in the rat. First, cells were categorised into three groups on the basis of soma and dendritic field size: Group RGA cells have large somata and dendritic field diameters, Group RGB cells have small somata and dendritic field diameters, whereas Group RGC cells have small to medium‐sized somata and medium‐to‐large dendritic field diameters. On the basis of dendritic field morphology and presence across the retina, each Group was then subdivided into subgroups. The significance of our results in terms of retinal ganglion cell function is discussed. J. Comp. Neurol. 385:309–323, 1997.

Vasopressor neurons in the rostral ventrolateral medulla of the rabbit

Neurons within the rostral ventrolateral medulla oblongata project directly to the intermediolateral column in the thoracolumbar spinal cord. This paper reviews evidence obtained from experiments in the rabbit regarding the anatomical connections and physiological, pharmacological and histochemical properties of these cells. The following hypotheses are discussed: an increase in the firing rate of these neurons leads to a rise in arterial pressure due to sympathetic vasoconstriction, but does not affect respiratory or other somatomotor activity; the bulbospinal pathway originating from these neurons is an essential component of the central pathways mediating baroreceptor and other cardiovascular reflexes; these neurons receive tonic GABAergic inhibitory inputs, which are not all of baroreceptor origin; many of these bulbospinal neurons synthesize adrenalize. The possible role of adrenaline in the function of these neurons is considered.

Differential expression of catecholamine biosynthetic enzymes in the rat ventrolateral medulla

Adrenergic (C1) neurons located in the rostral ventrolateral medulla are considered a key component in the control of arterial blood pressure. Classically, C1 cells have been identified by their immunoreactivity for the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH) and/or phenylethanolamine N‐methyltransferase (PNMT). However, no studies have simultaneously demonstrated the expression of aromatic L‐amino acid decarboxylase (AADC) and dopamine β‐hydroxylase (DBH) in these neurons. We examined the expression and colocalization of all four enzymes in the rat ventrolateral medulla using immunohistochemistry and reverse transcription‐polymerase chain reaction (RT‐PCR) analysis. Retrograde tracer injected into thoracic spinal segments T2–T4 was used to identify bulbospinal neurons. Using fluorescence and confocal microscopy, most cells of the C1 group were shown to be double or triple labeled with TH, DBH, and PNMT, whereas only 65–78% were immunoreactive for AADC. Cells that lacked detectable immunoreactivity for AADC were located in the rostral C1 region, and approximately 50% were spinally projecting. Some cells in this area lacked DBH immunoreactivity (6.5–8.3%) but were positive for TH and/or PNMT. Small numbers of cells were immunoreactive for only one of the four enzymes. Numerous fibres that were immunoreactive for DBH but not for TH or PNMT were noted in the rostral C1 region. Single‐cell RT‐PCR analysis conducted on spinally projecting C1 neurons indicated that only 76.5% of cells that contained mRNA for TH, DBH, and PNMT contained detectable message for AADC. These experiments suggest that a proportion of C1 cells may not express all of the enzymes necessary for adrenaline synthesis. J. Comp. Neurol. 432:20–34, 2001.

The pre-Bötzinger complex and phase-spanning neurons in the adult rat.

To characterise respiratory neurons in the pre-Bötzinger complex of adult rats, extracellular recordings were made from 302 respiratory neurons in the ventral respiratory group of sodium pentobarbitone anaesthetised adult rats. Neurons were located 0 to 1.6 mm caudal to the facial nucleus, and ventral to the nucleus ambiguus. The pre-Bötzinger complex comprised expiratory neurons (22%, 22/100), inspiratory neurons (37%, 37/100) and phase-spanning neurons (41%, 41/100). In contrast, 80% (125/157) of Bötzinger neurons were expiratory, and 80% (36/45) of rostral ventral respiratory group neurons were inspiratory. Rostrocaudally, the pre-Bötzinger complex extended about 400 microns, starting at the caudal pole of the nucleus ambiguus compact formation. The pre-Bötzinger complex was also characterised by a predominance of propriobulbar neurons (81%, 13/16). Furthermore, 68% (33/48) of expiratory-inspiratory neurons found were located within the pre-Bötzinger complex. The variety of neuronal subtypes in the pre-Bötzinger complex, including many firing during the expiratory-inspiratory transition is consistent with the hypothesis that this nucleus plays a key role in respiratory rhythm generation in the adult rat.

Morphology of retinal ganglion cells in a New World monkey, the marmoset Callithrix jacchus

We studied the morphology of retinal ganglion cells in a diurnal New World primate, the marmoset Callithrix jacchus. This species is of interest as a model for primate vision because it has good behavioural visual acuity, and the retina and subcortical visual pathways are very similar to those of Old World monkeys and humans. Ganglion cells were labelled by placing small crystals of the carbocyanin dye Dil into the optic fibre layer, or by intracellular injection of neurobiotin. Two main classes of ganglion cell were labelled. We call these Group A cells and Group B cells: they are respectively homologous to parasol and midget cell classes. Group A and Group B cells show similar patterns of dye coupling, dendritic stratification and dendritic field size as their counterparts in Old World monkeys and humans. A third group of cells, which we call Group C, is morphologically heterogeneous. Examples corresponding to wide‐field ganglion cell types described in Old World primates were encountered. One subgroup of C cells has a morphology very similar to that of the small bistratified (blue‐on) cell described in macaque retina, suggesting that this functional pathway is common to all primates. As for other New World monkeys, the marmoset shows a sex‐linked polymorphism of cone pigment expression, such that all males are dichromats and the majority of females are trichromats. No systematic differences in Group B cells were seen between male and female retinas, suggesting that trichromacy is not accompanied by specific changes in ganglion cell morphology.