Beyond weakness: Characterization of pain, sensory profile and conditioned pain modulation in patients with motor neuron disease: A controlled study.

BACKGROUND: Motor neuron diseases (MND) represent a group of disorders that evolve with inexorable muscle weakness and medical management is based on symptom control. However, deeper characterization of non-motor symptoms in these patients have been rarely reported. METHODS: This cross-sectional study aimed to describe non-motor symptoms in MND and their impact on quality of life and functional status, with a focus on pain and sensory changes. Eighty patients (31 females, 55.7 ± 12.9 years old) with MND underwent a neurological examination, pain, mood, catastrophizing and psychophysics assessments [quantitative sensory testing (QST) and conditioned pain modulation (CPM)], and were compared to sex- and age-matched healthy controls (HC). RESULTS: Chronic pain was present in 46% of patients (VAS =5.18 ± 2.0). Pain of musculoskeletal origin occurred in 40.5% and was mainly located in the head/neck (51%) and lower back (35%). Neuropathic pain was not present in this sample. Compared to HC, MND patients had a lower cold detection threshold (p < 0.002), and significantly lower CPM scores (4.9 ± 0.2% vs. 22.1 ± 0.2%, p = 0.012). QST/CPM results did not differ between MND patients with and without pain. Pain intensity was statistically correlated with anxiety, depression and catastrophism, and spasticity scores were inversely correlated with CPM (ρ = -0.30, p = 0.026). CONCLUSIONS: Pain is frequently reported by patients with MNDs. Somatosensory and CPM changes exist in MNDs and may be related to the neurodegenerative nature of the disease. Further studies should investigate the most appropriate treatment strategies for these patients. SIGNIFICANCE: We report a comprehensive evaluation of pain and sensory abnormalities in motor neuron disease (MND) patients. We assessed the different pain syndromes present in MND with validated tools, and described the QST and conditioned pain modulation profiles in a controlled design.

Early-life stress, corpus callosum development, hippocampal volumetrics, and anxious behavior in male nonhuman primates.

Male bonnet monkeys (Macaca radiata) were subjected to the variable foraging demand (VFD) early stress paradigm as infants, MRI scans were completed an average of 4 years later, and behavioral assessments of anxiety and ex-vivo corpus callosum (CC) measurements were made when animals were fully matured. VFD rearing was associated with smaller CC size, CC measurements were found to correlate with fearful behavior in adulthood, and ex-vivo CC assessments showed high consistency with earlier MRI measures. Region of interest (ROI) hippocampus and whole brain voxel-based morphometry assessments were also completed and VFD rearing was associated with reduced hippocampus and inferior and middle temporal gyri volumes. The animals were also characterized according to serotonin transporter genotype (5-HTTLPR), and the effect of genotype on imaging parameters was explored. The current findings highlight the importance of future research to better understand the effects of stress on brain development in multiple regions, including the corpus callosum, hippocampus, and other regions involved in emotion processing. Nonhuman primates provide a powerful model to unravel the mechanisms by which early stress and genetic makeup interact to produce long-term changes in brain development, stress reactivity, and risk for psychiatric disorders.

Contuse lesion of the rat spinal cord of moderate intensity leads to a higher time-dependent secondary neurodegeneration than severe one. An open-window for experimental neuroprotective interventions.

Secondary neurodegeneration takes place in the surrounding tissue of spinal cord trauma and modifies substantially the prognosis, considering the small diameter of its transversal axis. We analyzed neuronal and glial responses in rat spinal cord after different degree of contusion promoted by the NYU Impactor. Rats were submitted to vertebrae laminectomy and received moderate or severe contusions. Control animals were sham operated. After 7 and 30 days post surgery, stereological analysis of Nissl staining cellular profiles showed a time progression of the lesion volume after moderate injury, but not after severe injury. The number of neurons was not altered cranial to injury. However, same degree of diminution was seen in the caudal cord 30 days after both severe and moderate injuries. Microdensitometric image analysis demonstrated a microglial reaction in the white matter 30 days after a moderate contusion and showed a widespread astroglial reaction in the white and gray matters 7 days after both severities. Astroglial activation lasted close to lesion and in areas related to Wallerian degeneration. Data showed a more protracted secondary degeneration in rat spinal cord after mild contusion, which offered an opportunity for neuroprotective approaches. Temporal and regional glial responses corroborated to diverse glial cell function in lesioned spinal cord.

Transient and widespread astroglial activation in the brain after a striatal 6-OHDA-induced partial lesion of the nigrostriatal system.

The authors have previously described astroglial activation in the ipsilateral nigrostriatal system and ventral tegmental area following small doses of 6-hydroxydopamine (6-OHDA) injected unilaterally in the striatum. This article further evaluated astroglial reactivity in several brain regions after striatal 6-OHDA-induced punctate lesion in the nigrostriatal pathway. Adult male Wistar rats received a unilateral stereotaxical injection of the 6-OHDA (8 microg/4 microl) in the neostriatum and sacrificed 1 or 3 weeks later. Control animals received only solvent. Immunohistochemistry was employed for visualization of the tyrosine hydroxylase (TH), marker for dopamine cells, and glial fibrillary acidic protein (GFAP), marker for astrocytes. TH immunoreactive terminals disappeared in the striatum close to the injection site and a disappearance of a small number of a defined population of dopamine cell bodies was observed in the ipsilateral pars compacta of the substantia nigra (SNc). No dopamine lesion was detected in the contralateral nigrostriatal pathway. Astroglial reaction was seen close to the lesion in the neostriatum and in the ipsilateral SNc of the 1 week 6-OHDA lesioned rats. Specific stereological tools employing point intercepts and rotator, revealed an increased presence of reactive astrocytes in many forebrain regions like frontal, parietal and piriform cortex, septum, neostriatum and SNc, bilaterally, and also corpus callosum after 1 week of 6-OHDA injection. The astroglial activation was characterized by increases in the size of the cell body and/or processes. Astrocytic reaction was found only in the ipsilateral nigrostriatal pathway by 3 weeks of 6-OHDA, a slight activation also remaining in the ipsilateral septum and piriform cortex. Astrocytic reaction was seen in the solvent-injected rats only in the neostriatum close to the needle track. The transient widespread astroglial reaction observed in many brain regions following a striatal injection of 6-OHDA may represent a global paracrine trophic response in the brain.

FGF-2 and S100beta immunoreactivities increase in reactive astrocytes, but not in microglia, in ascending dopamine pathways following a striatal 6-OHDA-induced partial lesion of the nigrostriatal system.

Partial lesions were induced in rat midbrain dopamine ascending pathways by intrastriatal injection of 6-hydroxydopamine (6-OHDA), and after two weeks changes were observed in the immunoreactivities of S100beta, a calcium-binding protein, and basic fibroblast growth factor (FGF-2), which is neurotrophic. Semiquantitative microdensitometric image analysis revealed increased intensities of FGF-2 and S100beta immunostaining in putative glial profiles of the ipsilateral neostriatum, pars compacta (SNc) and reticulata (SNr) of the substantia nigra and ventral tegmental area (VTA). Double immunofluorescence and immunoperoxidase procedures, using antibodies against glial fibrillary acidic protein and OX-42, showed that these increased immunoreactivities were restricted to reactive astrocytes; they were not observed in reactive microglia. These results indicate that reactive astrocytes may exert paracrine trophic actions through S100beta and FGF-2 in the midbrain dopamine ascending pathways after striatal 6-OHDA treatment. Interactions between S100beta and FGF-2 may be relevant to neuronal maintenance and repair following dopamine injury.

Decreases in the expression of CGRP and galanin mRNA in central and peripheral neurons related to the control of blood pressure following experimental hypertension in rats.

Calcitonin gene-related peptide (alpha CGRP) and galanin (GAL) are peptides known to participate in central mechanisms of blood pressure control. Nonetheless, variations in the synthesis of the peptides in response to a hypertensive challenge are not well described, specially using a model, which allows acute and chronic analyses. In this study, we have employed in situ hybridization to analyse changes in mRNA expression of alpha CGRP and GAL in the nucleus tractus solitarii (NTS), hypothalamic paraventricular nucleus (PVN) as well as petrosal and nodose ganglia after aortic coarctation-induced hypertension in rats. Acute (2h) and chronic (3 and 7 days) analyses were performed in order to evaluate the involvement of both peptides in different periods of hypertension. The analysis of relative mRNA levels showed significant differences between sham-operated and aortic coarcted hypertensive rats. alpha CGRP mRNA expression was decreased 2h (40%) and 3 days (42%) in nodose and petrosal ganglia, respectively, after coarctation. No changes in CGRP mRNA signal were seen in the NTS and PVN in the analysed periods. GAL mRNA expression was decreased in the NTS (19%) and PVN (55%), 3 and 7 days, respectively, after coarctation-induced hypertension. No changes in GAL mRNA expression were observed in petrosal and nodose ganglia following aortic coarctation. Data suggest that alpha CGRP and GAL may participate in the mechanisms involved in the establishment/maintenance of hypertension induced by aortic coarctation. Acute changes might be involved with the adaptation to the hypertensive state, while changes at the chronic phase might be related to counteraction of hypertension.

Astroglial and microglial activation in the wistar rat ventral tegmental area after a single striatal injection of 6-hydroxydopamine.

Astroglial and microglial activation were analyzed in the ventral tegmental area (VTA) in adult male Wistar rats, after an unilateral striatal 6-hydroxydopamine (6-OHDA) injection. Different doses (8, 4, and 1 microg) of 6-OHDA were injected in the left side of the neostriatum; animals were sacrificed 22 days later. Control animals received an injection of the same volume of the solvent. The tyrosine hydroxylase (TH) positive dopamine cells, the glial fibrillary acidic protein (GFAP) immuno -labeled astrocytes, and the OX42 immunoreactive microglia were visualized by means of immunohistochemistry and quantified by stereologic methods employing the optical dissector and the point intercepts. The number and the density of TH immunoreactive cell bodies were decreased by 45% and 46%, respectively, in the sampled field of the ipsilateral VTA of 8 microg 6-OHDA injected rats. The GFAP immunohistochemistry revealed in the ipsilateral VTA increases the number and density of astroglial cells (154% and 166% of control, respectively) in the rats with a higher dose of the 6-OHDA, and also in the volume fraction of the astroglial processes after 8 microg (41% of control) and 4 microg (24% of control) of 6-OHDA. Increased number (76% of control) and density (77% of control) of OX42 microglial labeled profiles and microglial processes (51% of control) were found in the ipsilateral VTA of the 8 microg 6-OHDA injected animals. These results suggest that the retrograde degeneration of the mesostriatal dopamine pathways, induced by a striatal injection of 6-OHDA, leads to astroglial and microglial reactions in the VTA. The interaction between activated glial cells may be involved in the wounding and repair events in the partial lesioned system, and also in the trophic paracrine responses in the surviving VTA dopamine neurons.

Astroglial and microglial reaction after a partial nigrostriatal degeneration induced by the striatal injection of different doses of 6-hydroxydopamine.

Astroglial and microglial activation was analyzed in adult male Wistar rats after a unilateral striatal injection of different doses (8, 4 and 1 micrograms) of 6-hydroxydopamine (6-OHDA). Control animals received the injection of the same volume of the solvent. The rotational behavior was registered by a rotometer 24 and 72 hours, 7, 10, 14 and 22 days after lesion. Following, animals were sacrificed and the tyrosine hydroxylase (TH) positive dopamine cells, the glial fibrillary acidic protein (GFAP) immunolabeled astrocytes and the OX42 immunoreactive microglia were visualized by mean of immunohistochemistry and quantified by stereologic method employing the optical disector and the point intercepts. The apomorphine (0.5 mg/kg)-induced circling behavior was seen only after 8 micrograms of 6-OHDA from 72 hours postlesion until sacrifice. Decreases of the TH immunoreactive terminals and cell bodies were found in the sampled fields of the striatum and pars compacta of the substantia nigra (SNc), respectively, after 8 and 4 micrograms of 6-OHDA. The GFAP immunohistochemistry revealed increases in the number/density of astroglial cells in the ipsilateral neostriatum (137% of control) and ipsilateral SNc (83% of control) and also in the volumeal fraction of the astroglial processes in the ipsilateral neostriatum (30% of control) and ipsilateral SNc (38% of control) in the rats with higher dose of the neurotoxin. Increases in the number of OX42 microglial labeled profiles and in the volumeal fraction of microglial processes were found in the ipsilateral neostriatum (67% and 27%, respectively, of control) and ipsilateral SNc (100% and 50%, respectively, of control) in the 8 micrograms 6-OHDA injected rats. These results suggest that the retrograde degeneration induced by a intrastriatal injection of a small dose of the 6-OHDA leads to an astroglial and microglial reaction in the nigrostriatal dopamine pathway. The interaction between activated glial cells may be involved in the wounding and repair events in the partial lesioned nigrostriatal system as well as in the paracrine responses to surviving dopamine neurons.

Distant microglial and astroglial activation secondary to experimental spinal cord lesion.

This paper analysed whether glial responses following a spinal cord lesion is restricted to a scar formation close to the wound or they might be also related to widespread paracrine trophic events in the entire cord. Spinal cord hemitransection was performed in adult rats at the thoracic level. Seven days and three months later the spinal cords were removed and submitted to immunohistochemistry of glial fibrillary acidic protein (GFAP) and OX42, markers for astrocytes and microglia, as well as of basic fibroblast growth factor (bFGF), an astroglial neurotrophic factor. Computer assisted image analysis was employed in the quantification of the immunoreactivity changes. At the lesion site an increased number of GFAP positive astrocytes and OX42 positive phagocytic cells characterized a dense scar formation by seven days, which was further augmented after three months. Morphometric analysis of the area and microdensitometric analysis of the intensity of the GFAP and OX42 immunoreactivities showed reactive astrocytes and microglia in the entire spinal cord white and gray matters 7 days and 3 months after surgery. Double immunofluorescence demonstrated increased bFGF immunostaining in reactive astrocytes. The results indicated that glial reaction close to an injury site of the spinal cord is related to wounding and repair events. Although gliosis constitutes a barrier to axonal regeneration, glial activation far from the lesion may contribute to neuronal trophism and plasticity in the lesioned spinal cord favoring neuronal maintenance and fiber outgrowth.

Experimental models of partial lesion of rat spinal cord to investigate neurodegeneration, glial activation, and behavior impairments.

The article demonstrates two experimental models of spinal cord partial injury in rats: a contuse model promoted by the NYU impactor system and a partial hemitransection model achieved by a stereotaxic-positioned adjustable wire knife. By means of a defined impact weight (10 g) and a digital optical potentiometer linked to a computer, the impactor transferred and registered a moderate or a severe contusion to the rat spinal cord at a low thoracic level after dropping the weight from distances of 25 mm and 50 mm, respectively, to the dorsal surface of the exposed dura spinal cord. Impact curve was calculated and the parameters of the trauma, like impact velocity, cord compression distance and cord compression rates were obtained in order to demonstrate trauma severity. To promote partial hemitransection, rats were positioned in a spinal cord unit of a stereotaxic apparatus and lesion was made with the adjustable wire knife spatially oriented. By means of a computerized infrared motion sensor-home cage activity monitor and a noncomputerized evaluation of motor behavior using the inclined plane and the motor score of Tarlov tests, behavior was analyzed in an acute period postlesion. Rats were sacrificed and spinal cords were processed for routine staining to show neurons and for GFAP and OX42 immunohistochemistry to demonstrate glial cells. The tissue labelings were quantified using computer assisted stereology by means of an optical disector and microdensitometric image analysis by means of quantification of gray values of discriminated profiles. While partial hemitransection model favored a more accurate control of the lesion location, the contuse model allowed us to perform different degrees of lesion severity. A close correlation between behavioral impairment and severity of trauma was seen in the rats submitted to spinal cord contusion. The stereologic lesion index showed a correlation between severity of trauma and tissue damage by 7 days and demonstrated a time-dependent secondary degeneration after moderate but not after severe spinal cord contusion from 7 to 30 days after injury. Long-lasting activations of astrocytes and microglia seen by persisted increases in the specific mean gray values of immunoreactivities were also found in all levels of the white and gray matters of the partial hemitransected spinal cord until 3 months postinjury which can be related to wound/repair events and paracrine trophic support to spinal cord remaining neurons. The results showed that controlled partial lesions may provide an important toll to study trophism and plasticity in the spinal cord.

S100 immunoreactivity is increased in reactive astrocytes of the visual pathways following a mechanical lesion of the rat occipital cortex.

After demonstration of the paracrine action of glial neurotrophic factors, gliosis has also been considered to be related to neuronal trophism and plasticity rather than solely a repair event following brain injury. S100 is a Ca2+ binding protein, present mainly in astrocytes, that exerts paracrine trophic effects on several neuronal populations. This study analyses the presence of S100 protein by means of immunohistochemistry combined with stereology in the reactive glial cells of the rat visual pathways following a lesion of the visual cortex. Adult male Wistar rats were submitted to a unilateral aspiration of the occipital cortex or to a sham operation. One week later the rats were killed and their brain processed for immunochemistry. Single antibody immunohistochemistry was performed for the visualization of glial fibrillary acidic protein (GFAP, a marker for astrocytes), OX-42 (a marker for microglia) and S100 protein. Double immunofluorescence procedures were applied for co-localization of the S100/GFAP and S100/OX-42. An optical dissector, point interceptors and rotators were used to quantify the degree of glial activation and the changes in the S100 immunoreactivity. We observed an intense microglial and astroglial reaction in addition to an increased S100 immunoreactivity in the occipital cerebral cortex, geniculate nucleus and hippocampus ipsilateral to the lesion. In the ipsilateral superior colliculus, an intense astroglial activation was accompanied by an up-regulation of S100 immunoreactivity. Double-immunofluoresence revealed an increased S100 immunoreactivity in reactive astrocytes, but not in the reactive microglia. Evidence has therefore been obtained that after mechanical trauma, the astroglial S100 protein participates in the trophism and plasticity of the injured visual pathways.

The trophic factors S-100beta and basic fibroblast growth factor are increased in the forebrain reactive astrocytes of adult callosotomized rat.

S-100 is a calcium-binding protein that is predominantly found in astrocytes of the central nervous system. In the present study, we investigated the temporal and spatial changes of S-100beta immunoreactivity after a stereotaxic mechanical lesion of the adult rat corpus callosum performed with an adjustable wire knife. Rats were killed 7, 14 and 28 days after surgery. S-100beta immunoreactivity was found within the cytoplasm and processes of quiescent putative astrocytes that were observed throughout the gray and white matters of the forebrain of sham-operated rats. Following callosotomy, the S-100beta immunoreactive profiles showed increased size and thick processes, as well as increased amount of S-100beta immunoreactivity. Unbiased stereologic analysis revealed a sustained and widespread increase of the Areal Fraction of S-100beta immunoreactive profiles in the medial and lateral regions of the white matter of callosotomized rats at the studied time-intervals. In the cerebral cortex of callosotomized rats, the estimated total number of S-100beta immunoreactive profiles was also increased 7 and 14 days after the lesion. Since the cellular and temporal changes in S-100beta immunoreactivity were closely similar to those described for basic fibroblast growth factor (bFGF) following brain lesions, we co-localized the S-100beta and bFGF immunoreactivities after callosotomy. bFGF immunoreactivity was found in the nuclei of S-100beta immunoreactive glial profiles throughout the forebrain regions of the sham-operated rats. bFGF immunoreactivity was increased in the nuclei of reactive S-100beta immunoreactive putative astrocytes in the forebrain white matter and in the cerebral cortex of callosotomized rats. These results indicate that after transection of the corpus callosum of adult rats, the reactive astrocytes may exert paracrine trophic actions through S-100beta and bFGF. Interactions between S-100beta and bFGF may be relevant to the events related to neuronal maintenance and repair following brain injury.

Analysis of trophic responses in lesioned brain: focus on basic fibroblast growth factor mechanisms.

The actions of fibroblast growth factors (FGFs), particularly the basic form (bFGF), have been described on a large number of cells and include mitogenicity, angiogenicity and wound repair. The present review discusses the presence of the bFGF protein and messenger RNA as well as the presence of the FGF receptor messenger RNA in the rodent brain by means of semiquantitative radioactive in situ hybridization in combination with immunohistochemistry. Chemical and mechanical injuries to the brain trigger a reduction in neurotransmitter synthesis and neuronal death which are accompanied by astroglial reaction. The altered synthesis of bFGF following brain lesions or stimulation was analyzed. Lesions of the central nervous system trigger bFGF gene expression by neurons and/or activated astrocytes, depending on the type of lesion and time post-manipulation. The changes in bFGF messenger RNA are frequently accompanied by a subsequent increase of bFGF immunoreactivity in astrocytes in the lesioned pathway. The reactive astrocytes and injured neurons synthesize increased amount of bFGF, which may act as a paracrine/autocrine factor, protecting neurons from death and also stimulating neuronal plasticity and tissue repair.

Analysis of trophic responses in lesioned brain: focus on basic fibroblast growth factor mechanisms

The actions of fibroblast growth factors (FGFs), particularly the basic from (bFGF), have been described in a large number of cells and include mitogenicity, angiogenicity and wound repair. The present review discusses the presence of the bFGF protein and messenger RNA as well as the presence of the FGF receptor messenger RNA in the rodent brain by means of semiquantitative radioactive in situ hybridization in combination with immunohistochemistry. Chemical and mechanical injuries to the brain trigger a reduction in neurotransmitter synthesis and neuronal death which are accompanied by astroglial reaction. The altered synthesis of bFGF following brain lesions or stimulation was analyzed. Lesions of the central nervous system trigger bFGF gene expression by neurons and/or activated astrocytes, depending on the type of lesion and time post-manipulation. The changes in bFGF messenger RNA are frequently accompanied by a subsequente increase of bFGF immunoreactivity in astrocytes in the lesioned pathway. The reactive astrocytes and injured neurons synthesize increased amount of bFGF, which may act as a paracrine/autocrine factor, protecting neurons from death and also stimulating neuronal plasticity and tissue repair.

Myenteric neurons of the mouse small intestine. Morphometry and acetylcholinesterase activity.

1. The myenteric plexus of the small intestine of five C57BL/6J male 5-month-old mice was investigated in whole-mount preparations of the muscularis externa by Giemsa staining and by the acetylcholinesterase (AChE) histochemical technique. 2. The neuronal density was 20212 +/- 3038/cm2 (mean +/- SEM) in the duodenum, 21948 +/- 1488/cm2 in the jejunum, and 25048 +/- 2356/cm2 in the ileum. The difference in neuronal density between duodenum and ileum was statistically significant (P < 0.05). The total serosal surface area of the small intestine was about 30.80 +/- 2.90 cm2, and the total number of neurons was estimated at about 690,000. 3. The neuronal cell and nucleus profile areas ranged, respectively, from 23 to 325 microns 2 and from 6 to 95 microns 2 in the small intestine of the mice studied. There were no significant differences in any of the 3 regions in terms of average neuronal cell or nucleus profile areas. 4. For the histochemical demonstration of AChE, the "direct coloring" copper ferrocyanide method was used. AChE-positive nerve fibers were distributed in the myenteric plexus which was formed by a primary meshwork of relatively large nerve bundles and a secondary meshwork of finer nerve bundles. Most of the neurons of the plexus displayed AChE activity in the cytoplasm though the neurons presented different reaction intensities.(ABSTRACT TRUNCATED AT 250 WORDS)

Number, size and distribution of ganglion neurons in urinary bladder of rodents.

Whole-mount preparations of urinary bladders stained with a modified Giemsa technique were obtained from three rodent species (Guinea-pig, Calomys callosus and the C57/BLJ isogenic mouse) to identify and estimate the relative number and size of ganglionic neurons within the wall of the organ. The distribution of the ganglia was not uniform among the three species: ganglia were concentrated around the ureteral orifices in the Guinea-pig, they were scattered throughout the organ in Calomys callosus, and they were concentrated near the internal urethral orifice in the C57/BLJ mouse. In the Guinea-pig, the size of approximately 50% of the neurons lie in the range of 200 to 300 microns2. In Calomys callosus, 40% of the neurons lie in the range of 200 to 250 microns2, with 28% in the range of 50 to 100 microns2. For the C57/BLJ mouse, approximately 60% of the neurons have an area of 250 to 400 microns2.

Myenteric neurons of the mouse small intestine. Morphometry and acetylcholinesterase activity

1. The myenteric plexus of the small intestine of five C57BL/6J male 5-month-old mice was investigated in whole-mount preparations of the muscularis externa by Giemsa staining and by the acetylcholinesterase (AChE) histochemical technique. 2. The neuronal density was 20212 ñ 3038/cm² (mean ñ SEM) in the duodenum, 21948 ñ 1488/cm² in the jejunum, 25048 ñ 2356/cm² in the ilium. The difference in neuronal density between duodenum and ileum was statistically significant (P<0,05). The total serosal surface area of the small intestine was about 30.80 ñ 2.90 cm², and the total number of neurons was estimated at about 690,000. 3. The neuronal cell and nucleus profile areas ranged, respectively, from 23 to 325 µm² and from 6 to 95 µm² in the small intestine of the mice studied. There were no significant differences in any of the 3 regions in terms of average neuronal cell or nucleus profile areas. 4. For the histochemical demonstration of AChE, the "direct coloring" copper ferrocyanide method was used. AChE-positive nerve fibers were distributed in the myenteric plexus which was formed by a primary meshwork of relatively large nerve bundles and a secondary meshwork of finer nerve bundles. Most of the neurons of the plexus displayed AChE activity in the cytoplasm though the neurons presented different reaction intensities. 5. The results of the present study show that the myenteric plexus of the C57BL/6J mouse small intestine contains a large number of neurons which have different sizes and AChE activities

Number, size and distribution of ganglion neurons in urinary bladder of rodents

Whole-mount preparations of urinary bladders stained with a modified Giemsa technique were obtained from three rodent species (Guinea-pig, Calomys callosus and the C57/BLJ isogenic mouse) to identify andestimate the relative number and size of ganglionic neurons within the wall of the organ. The istribution of the ganglia was not uniform among the three species: ganglia were concentrated around the ureteral orifices in the Guinea-pig, they were scattered throughout the organ in Calomys callosus, and they were oncentrated near the internal urethral orifice in the C57/BLJ mouse. In the Guinea-pig, the size of approximately 50 percent of the neurons lie in the range of 200 to 300 microns2. In Calomys callosus, 40 percent of the neurons lie in the range of 200 to 250 microns2, with 28 percnet in the range of 50 to 100 microns2. For the C57/BLJ mouse, approximately 60 percent of the neurons have an area of 250 to 400 microns2

[Number and thickness of myelinic fibres of the phrenic nerve of young and aged rats]. / Número e espessura de fibras mielínicas do nervo frênico de rato albino jovem e senil.

The phrenic nerve of albino rats was studied for age changes in number of fibres, myelin sheath thickness and axon calibre. There is no significant morphological differences between nerves from young and aged rats and no difference with age was found in the number of fibres, myelin sheath thickness and axon calibre.