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Biomedical subjects

J Westman

Publications and source records attributed to J Westman.

At least 73 records · Page 4Linked to original sources

Synthesis of the methyl glycosides of a tri- and a tetra-saccharide related to heparin and heparan sulphate.

The methyl glycoside of a tetrasaccharide isolated from heparin, methyl O-(alpha-L-idopyranosyluronic acid)-(1-->4)-O-(2-acetamido-2-deoxy-alpha-D- glucopyranosyl)-(1-->4)-O-(beta-D-glucopyranosyluronic acid)-(1-->3)-O-beta-D-galactopyranoside disodium salt and a trisaccharide derivative thereof, methyl O-(alpha-L-idopyranosyluronic acid)-(1-->4)-O-(2-acetamido-2- deoxy-alpha-D-glucopyranosyl)-(1-->4)-O-beta-D-glucopyranosyluronic acid disodium salt, were synthesized using a block-type strategy. A suitable protected disaccharide block of iduronic acid and glucosamine (IdoA-GlcN) was used as a key intermediate for the syntheses and was glycosidated with a protected galactose derivative and a disaccharide block of glucuronic acid and galactose (GlcA-Gal) to give tri- and tetra-saccharide derivatives, respectively. Deprotection gave the target compounds.

Carbohydrate Sequence↗

Efflux of cholesterol from cholesterol loaded macrophages by incubation with synthetic HDL-particles.

Cells from the mouse monocyte/macrophage cell line J774A.1 were incubated with acetylated human low density lipoprotein for 2 days, resulting in an intracellular accumulation of mainly cholesteryl esters. These in vitro foam cell models were used to study the capability of synthetic HDL-particles to promote efflux of cholesterol. The synthetic HDL-particles were prepared from recombinant human pro-apolipoprotein A-I or human apolipoprotein A-I and phosphatidylcholine. Both types of reconstituted complexes were found to have a discoidal structure. A 24 h incubation of lipid loaded J774A.1 cells with these two types of discoidal complexes resulted in an equivalent and marked egress of cholesterol. The effect was the same whether the origin of phosphatidylcholine was egg yolk or soybean.

Animals↗

Acute systemic heat stress increases glial fibrillary acidic protein immunoreactivity in brain: experimental observations in conscious normotensive young rats.

The possibility that astrocytes participate in the pathophysiology of thermal brain injury caused by systemic heat exposure was examined in conscious young rats. The temporal and regional pattern of the astrocytic response to thermal injury was characterized by demonstrating the immunoreactivity of glial fibrillary acidic protein (GFAP) using monoclonal antibody and avidin-biotin complex technique. Exposure of conscious young animals to heat at 38 degrees C for 4 h in a biological oxygen demand incubator resulted in a marked increase of the GFAP immunoreactivity in specific brain regions as compared with the intact controls. The intensity of the increased GFAP immunoreactivity was mainly noted in pons, medulla and cerebellum, followed by thalamus, hypothalamus, hippocampus and caudate nucleus. The cerebral cortex of heat-exposed animals showed only a mild increase in GFAP immunoreactivity which was predominantly concentrated in cingulate, parietal and pyriform cortices. The immunostaining in general was seen in the perivascular glia, within the neuropil and the glia limitans. This increase in GFAP immunoreactivity was absent in animals exposed to the same ambient temperature (38 degrees C) for 1 h and 2 h, or at a lower temperature (36 degrees C) for 4 h. These results show that (i) astrocytes actively participate in the pathophysiology of heat stress, (ii) endogenous thermal brain injury elicits activation and hypertrophy of astrocytes ("reactive gliosis") depending on the magnitude and duration of the ambient heat stimulus, and (iii) the astrocytic reaction (observed as increased GFAP immunostaining) could be induced much more rapidly within a very short survival period of 4 h, not reported earlier.

Animals↗

Role of serotonin in heat adaptation: an experimental study in the conscious young rat.

The possibility that serotonin may play a role in heat adaptation was examined in a rat model of heat stress (HS). Subjection of animals to acute 4 h HS (but not to 1 h and 2 h duration) in a biological oxygen demand (BOD) incubator (relative humidity 50-55%, wind velocity 20-25 cm/s) resulted in profound hyperthermia (41.68 +/- 0.25 degrees C) associated with behavioural salivation and prostration. This experimental condition simulates the out door environmental conditions of Varanasi, India during the month of June. Measurement of serotonin (5-hydroxytryptamine, 5-HT) content in the plasma and brain showed a profound increase of the amine by 462% and 187% respectively from the control group. Morphological examination in these animals revealed marked cell changes in the brain at both light and electron microscopic level. Thus chromatolysis, shrunken and dark neurons along with a general sponginess of the gray and white matter was quite frequent in all the brain regions. At ultrastructural level, perivascular edema, damage to pre- and postsynaptic terminals, swollen neurons and glial cells, splitting of myelin and vacuolation was very common throughout the whole brain. The microvessels were mainly collapsed and a diffuse infiltration of lanthanum ion within the endothelium was common. Occasionally, lanthanum could also be seen within the basement membrane. However, the tight junctions were mainly intact. On the other hand, when animals exposed to chronic heat stress of 1 h duration at the same intensity for 7 days and then subjected to a 4 h HS on the 8th day, resulted in only minor symptoms. Thus these 4 h heat stressed animals did not exhibit hyperthermia or increased serotonin levels in plasma and brain as compared to the animals exposed to chronic HS of 1 h duration until alone 7th day. These heat adapted animals did not show any marked cell changes in the brain. However, the values of circulating 5-HT level and rectal temperature of these chronically heat stressed animals (for 1 h daily) were mild but significantly elevated on the 7th day as compared to the intact control animals. These results show that: 1. chronic HS of short duration induces heat tolerance; 2. the circulating 5-HT level some how appears to play an important role in the pathophysiology of HS; hand in the physiological mechanism(s) of secondary heat adaptation.

Adaptation, Physiological↗

Quantitative analysis of the feline dorsal column nuclei and their GABAergic and non-GABAergic neurons.

The gracile and internal and external cuneate nuclei of four adult cats were studied, using recently developed stereological techniques. The length, volume and position of the nuclei in relation to the level of obex were calculated, as well as the number of neurones, the neuronal density and volume of the three nuclei and different regions in the gracile and internal cuneate nucleus. Material processed for GABA immunocytochemistry was used in order to compare GABAergic and non-GABAergic neurones. The results demonstrate variations in the same nucleus in different animals, and in the nucleus of the left and right sides of the same animal. The same nucleus can vary up to 4 mm in its rostrocaudal position in relation to the obex. The mean sizes of the gracile, internal and external cuneate nuclei are 4.2, 8.4 and 5.6 mm3, respectively and their mean neuronal numbers are about 52,000, 76,000 and 33,000, respectively. The neuronal density was highest (12,907 cells/mm3) in the gracile, and lowest in the external cuneate nucleus (5987 cells/mm3). The external cuneate nucleus had a larger relative volume (7.9%) occupied by nerve cell bodies compared with the two medial nuclei (5.1% and 5.8%). In the gracile and internal cuneate nuclei, the GABAergic neurones constituted 28% and 25% of the whole population, respectively, while the external cuneate nucleus was devoid of such cells. All the nuclei contained GABA-positive boutons, however. The mean volume of the GABA-stained neurones in the gracile nucleus was 2319, and internal cuneate 3065 microns3, while the corresponding volume of unlabelled neurones in the gracile, internal and external cuneate nuclei was 3745, 8147 and 13318 microns3, respectively. When cyto-fibro-architectonic characteristics were used to subdivide the gracile and cuneate nuclei into rostral, middle and caudal regions, and the data of the three compartments compared, it was found that in both nuclei the middle region had the highest neuronal packing density, and the caudal region the largest mean nerve cell volume.

Animals↗

Ascending afferents to the lateral cervical nucleus are enriched in glutamate-like immunoreactivity: a combined anterograde transport-immunogold study in the cat.

To investigate whether glutamate (Glu) may be a transmitter in terminals of ascending afferents to the lateral cervical nucleus (LCN), these terminals were identified by anterograde transport of wheatgerm agglutinin-horseradish peroxidase from the spinal cord, and their content of Glu-like immunoreactivity (Glu-LI) was assessed at the ultrastructural level by the immunogold technique. The gold particle density over the peroxidase-positive terminals of the spinocervical tract (SCT) was significantly higher (by a factor of 2.44) than over a reference terminal population containing flattened or pleomorphic vesicles. Further, LCN neurons were densely labeled by the Glu antiserum, although the gold particle density over neuronal cell bodies was not as high as in the SCT terminals. Previous investigations have shown enrichment of Glu-LI in putative glutamatergic terminals in other parts of the CNS. Hence, the present observations indicate that Glu may be a transmitter in the synapses between SCT terminals and LCN neurons. The cell body labeling in the LCN is more difficult to interpret because of possible interference of metabolic pools of glutamate.

Afferent Pathways↗

Light microscopical study of dendrites and perikarya of interneurones mediating la reciprocal inhibition of cat lumbar alpha-motoneurones.

Interneurones which mediate disynaptic inhibition from la muscle spindle afferents of the quadriceps nerve to lumbar alpha-motoneurones were stained with intracellular injection of horseradish peroxidase. Seven best stained and most satisfactorily preserved cells were selected for analysis, and the light microscopic morphology of their cell bodies and dendrites were quantitatively investigated in parasagittal sections. The perikarya were located dorsal or dorso-medial to the motoneurones; they had mean diameters of 51 x 27 microns and a mean volume of 35,820 microns 3. The cells had 3 to 7 dendrites, which were arranged asymmetrically around the parent somata. The dendrites extended mainly in the dorso-ventral direction, in which the mean tip to tip distance for each cell was 1742 microns. The dendrites had few spines and they branched almost only in bifurcations. On the average, each process divided 3.5 times and in each cell they gave rise to 14.9 branching points as well as a total combined length of more than 7000 microns. Primary dendrites had a mean length of 193 microns which was generally shorter than the lengths of the branches of higher order. A more detailed analysis of two cells revealed the mean width of primary dendrites to be 5.6 microns while that of the 5th order processes was 1.5 microns. The mean tapering of individual dendritic branches per unit length was 17%, being somewhat more pronounced for the distally located segments, while at branching points the sum of daughter processes approximately equalled the diameter of the parent process.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Increased 5-hydroxytryptamine immunoreactivity in traumatized spinal cord. An experimental study in the rat.

The possibility that serotonin (5-hydroxytryptamine, 5-HT) is involved in the early tissue reactions occurring in spinal cord trauma was examined in a rat model using an immunocytochemical technique. The injury was made in the form of a 5-mm long and 2.5-mm wide lesion of the right dorsal horn at the level of T10-11. Injured rats, pretreated with the 5-HT synthesis blocking agent, p-chlorophenyl alanine (p-CPA) were compared with untreated injured controls and the animals were allowed to survive for 5 h. The distribution of 5-HT was examined in proximal and distal cross- sections of the cord, located 2 and 5 mm away from the injury. Normal rats showed immunoreactive material in nerve cell processes and in a few nerve cell bodies of the ventral horns. The trauma to the spinal cord caused a marked increase in 5-HT immunoreactivity in the segments located 2 mm proximal and distal to the injury, particularly in the ipsilateral ventral horn. The segment located 5 mm distal to the lesion showed a similar increase in immunoreactivity but it was apparently less pronounced in the corresponding proximal segment. Treatment with p-CPA markedly reduced the trauma-induced increase in 5-HT immunoreactivity in all the segments. These immunohistochemical findings were in line with the changes in the contents of 5-HT measured biochemically in corresponding spinal cord segments. At the onset of the trauma to the spinal cord 5-HT is thus present in the tissue, mainly in the form of 5-HT-containing nerve cell processes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Permeabilization of hepatocytes by a saponin and the effects of dextran.

The process by which a saponin derived from Gypsophila plants permeabilizes rat hepatocytes was studied. When monolayer cultures were incubated with 25 micrograms/ml saponin in phosphate buffered saline, the amount of cell-bound saponin increased for at least 90 min. Release of intracellular K+ started immediately, with a t1/2 of about 5 min. ATP and lactate dehydrogenase (LDH) began to appear in the medium only after lag periods of 10 to 20 min with t1/2s of 20 to 30 min. Removing the saponin from the medium after 15 min stopped any further release of ATP and LDH, showing that increased permeability to small ions alone does not lead to lysis by colloid osmotic pressure. However, the lysis that occurred upon 30 min continuous incubation with the saponin could be inhibited (delayed) by the addition of an osmotically active compound - a dextran. These results indicate that increasing binding of the saponin destabilizes the plasma membrane so that it will rupture from the colloid osmotic pressure.

Adenosine Triphosphate↗

Light and electron microscopical studies of the substance P innervation of the dorsal column nuclei and the lateral cervical nucleus in the primate.

In 2 monkeys of the species Macaca fascicularis the three dorsal column nuclei and the lateral cervical nucleus have been investigated immunocytochemically with antiserum against Substance P. The Substance P labeling was widely spread but rather sparse. It occurred in small structures of the size of boutons in the gracile, main cuneate and lateral cervical nucleus. The most intensive labeling of the gracile nucleus was found in the dorsal border of the nucleus and the lateral part of the caudal division. At the border between the spinal trigeminal nucleus and the triangular part of the cuneate nucleus Substance P labeling was also increased but mainly localized to the former nucleus. In the pars rotunda and the caudal part of the main cuneate nucleus there was a more intense labeling laterally especially at the obex level. In the lateral cervical nucleus Substance P positive structures seemed evenly spread and somewhat more numerous than in the gracile and main cuneate nucleus. Electron microscopy demonstrated Substance P positive boutons, which were fairly large and mostly in synaptic contact with dendrites. The results from the different nuclei in the monkey were compared with the results of similar investigations in the cat. It is concluded that there are important species differences especially on the light microscopical level in the lateral cervical nucleus. Thus Substance P terminals are evenly spread over the nucleus in the monkey whereas in the cat those structures are concentrated to the ventromedial region.

Afferent Pathways↗

Light and electron microscopical studies of the GABA innervation of the dorsal column nuclei and the lateral cervical nucleus in the primate species Macaca fascicularis and Papio anubis.

In 4 monkeys of the species Macaca fascicularis (2 animals) and Papio anubis (2 animals) the three dorsal column nuclei and the lateral cervical nucleus have been investigated immunocytochemically with antiserum against gamma-aminobutyric acid (GABA). Light microscopic studies demonstrated the presence of GABA-positive cells in the gracile nucleus, the internal cuneate nucleus and the lateral cervical nucleus but not in the external cuneate nucleus. Although labeled cells seemed fairly evenly spread in the nuclei there was an increased amount between the clusters of the internal cuneate nucleus and in the border zone between the gracile and the cuneate nucleus. Electron microscopical investigation showed GABA labeling in fairly small neurons with relatively large cell nuclei and low somatic bouton covering. GABA-positive terminals with rounded synaptic vesicles were present in all the investigated nuclei also the external cuneate one. No apparent difference in number of such boutons in the different nuclei or parts of the nuclei was found. GABA-positive boutons mostly synapsed with dendrites but in the dorsal column nuclei also with other larger boutons. Axosomatic contacts between labeled terminals and neuronal perikarya were more common in the lateral cervical nucleus than in the dorsal column nuclei. The results from the different nuclei in the monkey were compared with the results of similar investigations in the cat. It is concluded that there are important species differences especially on the light microscopical level in the lateral cervical nucleus. Thus the GABA labeled cells is rather evenly spread over the nucleus in the monkey whereas in the cat they are concentrated to the ventromedial region.

Animals↗

GABA-immunoreactive neurons and terminals in the lateral cervical nucleus of the cat.

Previous findings have indicated the presence of local circuit neurons in the lateral cervical nucleus (LCN). An immunohistochemical study with gamma-aminobutyric acid (GABA) antiserum was therefore performed both to investigate whether GABA-immunoreactive neurons are present in the LCN, and if so, to compare their characteristics with those previously assigned to probable internuncial neurons in the nucleus. The fine structure and synaptology of GABA-positive boutons in the LCN were also studied. Transversely cut sections from the upper cervical spinal cord of three cats were processed for GABA immunohistochemistry with the free-floating PAP technique. On light microscopic examination immunoreactive neurons were observed within the ventromedial half of the LCN. Their total number was estimated to be 42.5 +/- 11.7 in the entire LCN on one side of the cervical spinal cord, but this may have been an underestimation, as the penetration by the antisera was limited. The labeled neurons were small and had a relatively large nucleus and a low bouton covering ratio. In their number, localization, and ultrastructural appearance the GABA-positive neurons closely resembled the population of neurons previously suggested to be local circuit neurons. Immunoreactive bouton-sized puncta were scattered throughout the LCN. Ultrastructural examination showed labeled terminals with a mean sectional area of 0.85 micron 2 and a relatively high density of synaptic vesicles. The vast majority of GABA-positive terminals were in contact with dendrites and only a minority had synaptic contact with cell bodies. No axoaxonal synapses were observed. The GABA-positive boutons probably derive at least partly from the observed GABA-positive neurons, but there is also a possibility of extrinsic GABAergic input.

Animals↗

Lysosomes and pancreatic islet function. Time course of insulin biosynthesis, insulin secretion, and lysosomal transformation after rapid changes in glucose concentration.

The aim of this study was to estimate the time course of lysosomal transformations associated with crinophagy, i.e., the degradation of insulin within lysosomes, in the beta-cells of pancreatic islets. Primary and secondary lysosomes were identified in mouse islet beta-cells and subjected to ultrastructural morphometry. Islets from an in situ preparation were compared with isolated islets incubated in vitro. Under the in vitro conditions, the islets were initially exposed to 28 or 3.3 mM glucose for 24 h. Then the glucose concentration was rapidly changed to 3.3 and 28 mM glucose, respectively, and the islets were incubated for up to an additional 24 h. The beta-cell lysosomes were analyzed and related to alterations in insulin biosynthesis and secretion and islet insulin content after the rapid change in glucose concentration. In vivo, the beta-cell lysosomal population was predominantly composed of secondary lysosomes, which frequently contained secretory granule cores. After the initial 24-h period at 3.3 mM glucose, the volume density and the average volume of the secondary beta-cell lysosomes were increased, suggesting increased crinophagic activity. The mean diameter of the primary beta-cell lysosomes was decreased after 24 h at either 28 or 3.3 mM glucose. The change in glucose from 28 to 3.3 mM resulted in alterations in insulin biosynthesis and secretion, leading to an accumulation of insulin within the beta-cells. Lysosomal transformations suggestive of increased crinophagy were observed 24 h after the alteration in glucose concentration. The change from 3.3 to 28 mM glucose resulted in a parallel increase in insulin biosynthesis and secretion without a change in islet insulin content.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Quantitative ultrastructural study of boutons of ascending afferents to the feline lateral cervical nucleus.

A quantitative ultrastructural study has been performed of 300 labelled and the same number of unlabelled boutons in the feline lateral cervical nucleus (LCN) of 5 adult cats after cervical or lumbar injections of lectin-conjugated horseradish peroxidase and histochemical reaction with tetramethylbenzidine. On an average the labelled boutons were slightly elongated, had areas of 2.25 micron 2, mitochondrial volume fractions of 11.2% and densities of synaptic vesicles of 35.4.micron-2. The synaptic vesicles were round to flattened with a mean length of 60 nm and length/width ratios of 1.21. The labelled boutons were significantly larger and had significantly lower densities of synaptic vesicles than the unlabelled boutons. The labelled boutons constituted about 14% of the bouton volume of the investigated areas of the LCN. Most of them were axo-dendritic and about 16% synapsed with cell bodies with ultrastructural characteristics similar to the cervico-thalamic projection neurones of the LCN. The number of boutons labelled after the injections comprising 3-5 adjacent segments of the spinal cord was calculated to 6.8 X 10(6). Based on the assumption that they represented terminals of spinocervical tract cells, it was calculated that each of these cells in the lumbar cord gives rise to an average of 4400 boutons in the LCN.

Afferent Pathways↗

Somatosensory projection to the mesencephalon: an anatomical study in the monkey.

The terminal areas and cells of origin of the somatosensory projection to the mesencephalon in the monkey were investigated by the intraaxonal transport method. Following injection of wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) into the spinal enlargements, the lateral cervical nucleus (LCN), the dorsal column nuclei (DCN), or the spinal trigeminal nucleus, anterograde labeling was observed in several regions of the mid-brain. (1) Injection of tracer into the spinal enlargements resulted in dense terminal labeling in the parabrachial nucleus (PBN) and the periaqueductal gray matter (PAG); moderate termination was observed in the intercollicular nucleus (Inc), the intermediate and deep gray layers of the superior colliculus (SGI, SGP), the posterior pretectal nucleus (PTP), and the nucleus of Darkschewitsch (D); and scattered terminal fibers were seen in the cuneiform nucleus (CNF) and the pars compacta of the anterior pretectal nucleus (PTAc). The projections from the cervical enlargement to PAG, Inc, and the superior colliculus terminated more rostrally than those from the lumbar segments, indicating a somatotopic organization. (2) Terminal labeling after injection of tracer into LCN was found mainly in Inc, SGI, and SGP, but sparse labeling was also observed in the nucleus of the brachium of the inferior colliculus (BIN), PAG, PBN, PTP, and D. (3) The projection from DCN terminated densely in the external and pericentral nuclei of the inferior colliculus (ICX, ICP), Inc, SGI, SGP, PTP, PTAc, the nucleus ruber, and D, and weak terminal labeling was seen in BIN, PAG, and PBN. Comparisons of the anterograde labeling following injections involving both the gracile nucleus and the cuneate nucleus with that after injection restricted to the gracile nucleus alone suggested a somatotopic termination pattern in Inc, the superior colliculus, and the pretectal nuclei. (4) The patterns of projection from the laminar and alaminar parts of the spinal trigeminal nucleus differed: injection of tracer into the caudal part of the alaminar spinal trigeminal nucleus (nucleus interpolaris) resulted in dense anterograde labeling in SGI and SGP, moderate termination in Inc, and minor projections to PBN, PAG, and PTP, whereas after tracer injection into the laminar trigeminal nucleus (nucleus caudalis) terminal labeling was present only in PBN and PAG. Following injection of tracer into the midbrain terminal areas retrogradely labeled neurons were found in the spinal cord, LCN, DCN, and the spinal trigeminal nucleus, with the majority of labeled cells situated on the side contralateral to the injection site.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

Postnatal development of the feline lateral cervical nucleus: I. A quantitative light and electron microscopic study.

With the aim of obtaining some basic information for future developmental studies, the lateral cervical nucleus (LCN) was investigated in 32 kittens of different ages by electron microscopic and stereologic methods. Corresponding light microscopic measurements of neuronal and nuclear profiles and of the total LCN volume were also performed. The total LCN volume increased sixfold between the ages of 12 hours and 120 days, the most rapid increase occurring during the first month. The neuronal size was fairly constant up to the age of 9 days, whereafter it showed greater variation. The mean profile area increased rapidly during the second week and then more slowly. The relative volume of boutons increased significantly between birth and the age of 34 days and then decreased slightly up to 120 days postnatally. The total bouton volume showed a steady increase, which was most pronounced between the ages of 9 and 34 days. The relative dendritic volume decreased during the 120 days of observation, whereas the total volume of dendrites increased up to the age of 92 days and then decreased. The total volume of glial cells increased during the 120-day observation period, as did both the relative and total volumes of myelinated axons. The changes in the relative volumes of mitochondria in boutons and dendrites were very similar, with increases that were most marked between the ages of 9 and 34 days and between 92 and 120 days.

Animals↗

The projection to the mesencephalon from the sensory trigeminal nuclei. An anatomical study in the cat.

The terminal areas and the cells of origin of the projection from the sensory trigeminal nuclei to the mesencephalon were investigated, using the method of anterograde and retrograde transport of horseradish peroxidase or wheat germ agglutinin-horseradish peroxidase conjugate. Injection of tracer into the nucleus interpolaris or nucleus oralis (in the latter cases with involvement of the nucleus principalis) resulted in dense anterograde labeling in the deep and intermediate gray layers of the contralateral superior colliculus, extending throughout the rostrocaudal extent of the colliculus with the exception of its caudalmost part, which was not labeled. Minor projections to the intercollicular nucleus, posterior pretectal nucleus and nucleus of Darkschewitsch were found. Injection of tracer into the nucleus caudalis yielded a completely different result; terminal labeling in the midbrain was now present only in the periaqueductal gray matter, in its rostral and middle parts. The retrograde labeling observed after injection of tracer into the midbrain terminal areas showed that the cells of origin were located mainly in the alaminar spinal trigeminal nucleus, and the highest density of labeled neurons was found in the rostral part (subnucleus y) of the nucleus oralis. The retrograde labeling in the nucleus principalis was very sparse and almost exclusively involved peripherally located neurons. In the nucleus caudalis the overwhelming majority of the retrogradely labeled neurons were situated in its marginal layer. The functional implications of the above observations are discussed in relation to the findings in previous studies of the projections from the dorsal column nuclei and spinal cord to the midbrain. The combined results suggest that the trigeminal projections to the superior colliculus may be involved in the mechanisms of orientational behavior. The observation that the projection to the periaqueductal gray matter originates in the marginal layer suggests that it transmits information related to noxious stimuli.

Afferent Pathways↗

Different neuron populations in the feline lateral cervical nucleus: a light and electron microscopic study with the retrograde axonal transport technique.

A comparative light and electron microscopic study was made of the two major neuron populations in the feline lateral cervical nucleus (LCN) that have an ascending projection--namely, the cervicothalamic and cervicomesencephalic neurons. The small population of neurons in the LCN that remained unlabelled after extensive injections into all known areas of LCN termination was also investigated. The intra-axonal retrograde tracing technique was employed and three different tracers were used: native horseradish peroxidase, horseradish peroxidase conjugated to wheat germ agglutinin, and cholera toxin subunit B (CTb). The diaminobenzidine and tetramethylbenzidine methods were used for the peroxidase reaction, and an immunocytochemical staining method with monoclonal antibodies was employed to localize CTb. The light microscopic cell count was performed on plastic-embedded semithin (2 micron) sections, and a section-embedding technique was used for sampling regions of interest for electron microscopy. The total number of neurons in the LCN was calculated to be 8,300, which is higher than numbers reported from other studies on frozen sections. It was concluded from control experiments that many neurons fail to counterstain in frozen sections. The average proportion of labelled cervicothalamic neurons was 94%, which accords with earlier reports. The frequency of labelled neurons projecting to the midbrain was 25-49%, depending on the method used. The highest number of labelled neurons was found with the immunocytochemical method with use of CTb as a tracer. In the electron microscopic study, the cell area, form factor, nuclear area, bouton covering ratio, mean appositional length of boutons, and bouton density were measured. There were no significant differences in these respects between the cervicothalamic and cervicomesencephalic neurons, but the unlabelled neurons differed significantly from the labelled ones, with a smaller somal area, a lower bouton covering ratio, and a larger relative nuclear area. The ascending neurons were scattered over the entire LCN, but the neurons that remained unlabelled after the extensive injections was found in its ventromedial part. It is suggested that these are interneurons. In the light microscopic study of the animals injected with CTb, the ventromedial part of the LCN displayed labelled structures resembling terminals. Electron microscopic examination of the sections revealed boutons synapsing on unlabelled neurons and dendrites. Occasionally labelled neurons and dendrites were also contacted.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗