Paragangliomas of the craniocervical region

Summary An immunohistochemical study on tyrosine hydroxylase (TH), a rate-limiting enzyme in the catecholamine synthesizing pathway, was made on three cranioncervical region paragangliomas, two of which showed metastases to the cervical lymph nodes. In all of the original tumors, the majority of tumor cells showed positive immunostaining for TH of variable intensity in their cytoplasm regardless of their cytological features such as cellular and nuclear pleomorphism. The finding suggests that most tumor cells are capable of production of catecholamines and are derived from chief cells in the normal paraganglia. In cervical lymph nodes, however, no positive immunostaining for TH was observed in metastatic tumor cells, in contrast with the findings in the original tumors. The absence of TH immunoreactivity in metastatic tumor cells appears to be noteworthy in considering their malignant potential. Application of the TH immunohistochemistry to further cases appears important for the better understanding of this neoplasm, a catecholamine-producing tumor.     

Esthesioneuroepithelioma: a tumor of true olfactory epithelium origin

Summary A case of esthesioneuroepithelioma was investigated ultrastructurally and immunohistochemically, using antibodies against neurofilament protein (NFP), glial fibrillary acidic protein (GFAP), keratin, neuron-specific enolase (NSE), S-100 protein (S-100), and tyrosine hydroxylase (TH). The tumor initially manifested as an epidural mass in the anterior cranial fossa in a 64-year-old man, and about 31/2 years later, autopsy further revealed extensive metastases to the lymph nodes of the neck and thoracic cavity. In the cranial and nasal cavities, the tumor was composed of fairly uniform, ill-defined cells arranged in nests which were surrounded by a fibrovascular stroma. These histological features were reproduced in the metastatic tumor nodules with frequent occurrence of tubular arrangements of the tumor cells. Ultrastructurally, two different cell types were well recognized by their characteristic morphological features, which were reminiscent of sensory neurons and sustentacular cells of the olfactory epithelium. No dense-cored secretory granules were observed in the tumor cells. Immunohistochemically, the tumor showed a variable number of cells positive for NFP, keratin, NSE and S-100. NFP was present in a relatively small number of cells, which were found diffusely in the nests. Keratin was observed in the cells mainly located at the periphery. NSE-positive cells tended to form irregular clusters in the center. A few S-100-positive cells were found, without any particular arrangement. These findings indicated that the present tumor, which actually arose in the superior nasal cavity, consisted of cells differentiating in at least two distinct directions, neuronal and epithelial, and strongly suggested that the tumor was of true olfactory epithelium origin, or more precisely, derived from the bipotential, undifferentiated basal cells of this epithelium.     

Development of DARPP-32-positive parts of fetal pig ganglionic eminence and ventral mesencephalon in organotypic slice co-cultures

Neurons from the fetal pig dopaminergic ventral mesencephalon (VM) and basal ganglia anlage (the ganglionic eminence) were co-cultured as organotypic slice cultures to study the development of the two interconnected brain areas. During a short developmental period (E35-E42), a groove separates the ganglionic eminence into a lateral and a medial part. This was used (a) to study the developmental expression of the striatal marker protein, dopamine and adenosine 3,5-monophosphate regulated phospho-protein (DARPP-32) in the two parts and (b) to compare innervations of the two parts by tyrosine hydroxylase (TH)-positive, dopaminergic fibers from co-cultured slices of the ventral mesencephalon. DARPP-32 expression was more extensive and dense in cultures of the lateral part of the striatal anlage than the medial part. The DARPP-32-positive areas moreover overlapped with areas rich in acetylcholine esterase (AChE) and were the preferred target areas for TH-positive fibers from the co-cultured VM.     

The development of the chromaffin cell lineage from the neural crest

Chromaffin cells are neuroendocrine cells, which are highly specialized for the synthesis and release of multiple hormones. Like sympathetic neurons, which are essential, inter alia, for neural control of vascular tone, they are derivatives of the neural crest, a transient structure at the dorsal surface of the embryonic neural tube. Chromaffin cells and sympathetic neurons have many features in common, but are also distinct in several respects. This review provides a summary of similarities and differences regarding the development of chromaffin cells and sympathetic neurons, viewed from molecular and morphological perspectives. Two major, still not finally settled issues, are whether (1) the two related cell types arise from one common or two separate cell lineages of delaminating neural crest cells, (2) in the former case when does lineage segregation occur, and what are the molecules underlying their phenotypic diversification.     

Dopaminergic terminals form synaptic contacts with enkephalinergic striatal neurons in pigeons: an electron microscopic study

Medium spiny projection neurons of the striatum consist of two major neuropeptide-specific types, one type containing substance P and another type containing enkephalin. Both of these types have been shown to receive dopaminergic input onto their perikarya and proximal dendrites. However, whether each of these types receives direct dopaminergic input onto distal dendritic shafts and onto dendritic spines has not been explored in depth. In the present study, we used electron microscopic immunohistochemical double-label techniques to examine the synaptic organization of dopaminergic input onto enkephalin-positive (ENK +) striatal neurons in pigeons, in whom ENK + striatal perikarya, dendritic shafts and spines can be readily labeled. Antibodies against tyrosine hydroxylase were used to label dopaminergic terminals using a silver-intensified immunogold method. ENK + neurons were labeled using diaminobenzidine. We found that dopaminergic terminals make appositions and form symmetric synapses with the perikarya, dendritic shafts, and dendritic spine necks of ENK + striatal neurons. Thus, nigral dopaminergic neurons provide a monosynaptic input onto ENK + striatal neurons in a manner similar to that described previously by us for substance P-positive striatal medium spiny neurons.     

Immunohistochemical localization of a 40-kDa catecholamine regulated protein in the nigrostriatal pathway

The 40 kDa catecholamine regulated protein (CRP40) has been shown to covalently bind catcholamines in vitro. In this report we provide evidence for CRP40 localization in the dopaminergic nigrostriatal pathway. Using double labeling immunohistochemistry, CRP40 was detected in the majority of substantia nigra pars compacta and striatal neurons. In addition, CRP40 was also present in tyrosine hydroxylase immunonegative neurons. Subcellular localization of CRP40 shows a predominant nuclear presence in striatal neurons while distinct outlining of the nucleus and cell body staining were seen more readily in nigral neurons. These findings suggest that CRP40 may have multiple functions in a variety of neurons in the central nervous system.     

The lateral ganglionic eminence is the origin of cells committed to striatal phenotypes: neural transplantation and developmental evidence

In order to determine whether the lateral ganglionic eminence (LGE) of the fetal telencephalon is the primary source of striatal precursors in striatal transplants and tissue cultures, cells derived exclusively from the LGE of fetal rat brains were transplanted into the quinolinic-acid-lesioned striatum of adult rats. After 2–3 months they produced grafts that were almost entirely AChE-positive as well as DARPP-32-, TH-, and calbindin-immunoreactive. The grafts were integrated into the host striatum so that host corticofugal fiber tracts interdigitated with graft tissues similar to the way they penetrate the gray matter of the normal striatum. Fast Blue dye injected into the ipsilateral globus pallidus of LGE grafted produced retrogradely labeled neurons within the grafts, but Fluorogold dye injected into the ipsilateral substantia nigra did not. In a separate experiment using DARPP-32-immunohistochemistry as a striatal marker, fetal (E16) and neonatal (P2) rat brains showed DARPP-32 immunoreactivity in the LGE but not in the adjacent medial ganglionic eminence (MGE). In summary, both fetal LGE cells and LGE grafts express specific striatal markers, and LGE grafts integrate into the host striatum and innervate the major striatal efferent target within the host brain. These data suggest that the LGE is the origin of cells committed to striatal phenotypes in the developing brain.     

A new chromogen for use in HRP-tract tracing and double-label immunocytochemistry

A new chromogen, Indophane Blue (IB) was tested for use in HRP-tract tracing and double-label immunocytochemistry. Although TMB was slightly more sensitive and delineated individual labeled fibers better than IB in tract-tracing experiments, there were two advantages to using IB over TMB. First, the IB reaction could be performed at a pH of 6.0, rather than the pH of 3.3 required for TMB, resulting in much better preservation of tissue. Second, there was no crystalline artifact as has been observed with TMB. In the double-label immunocytochemistry experiments, the combination of diaminobenzidine for the first chromogen and IB as the second, provided excellent contrast.     

Primary rhabdomyosarcoma combined with chronic paragonimiasis in the cerebrum: a necropsy case and review of the literature

Summary A necropsy case of a primary rhabdomyosarcoma with chronic paragonimiasis in the cerebrum of a 68-year-old man is reported. The clinical data showed a right hemiplegia and dysarthria which became lethal in 6 months even though operation and radiation therapy were performed. Computed tomography revealed a large low-density area associated with the peripheral enhancement in the left basal ganglia, and multiple conglomerated calcified masses in the left temporal and occipital lobes. Biopsied and necropsied materials of the tumor in the basal ganglia was reddish brown in color and histologically was composed of purely mesenchymal derivatives with both embryonal and mature striated muscle cells but neither neuronal nor glial elements. Some of the tumor cells with extending slender cytoplasms showed obvious cross striations at the light and electron microscope levels and immunohistochemical reactivity for myoglobin. All tumor cells were also positive for vimentin, but not for glial fibrillary acidic protein. The clinical and necropsy findings revealed no primary lesion anywhere but in the brain. In addition, numerous dead oval eggs ofParagonimus westermani were found in many cystoid lesions encapsulated by thick connective tissues with calcification and/or ossification. Clinicopathological features of 24 cases of primary rhabdomyosarcoma of the central nervous system reported in the literature are reviewed briefly. The histogenesis of this tumor are discussed together with comments on cerebral paragonimiasis.     

A combined behavioral and morphological study on the effects of fetal asphyxia on the nigrostriatal dopaminergic system in adult rats

Fetal asphyxic insults in the brain are known to be associated with developmental neurological problems like neuromotor disorders. However, little is known about the long-term consequences of fetal asphyxia (FA). For that reason, the present study investigated the long-term effects of FA on motor behavior and dopaminergic circuitry. FA was induced at embryonic day 17 by 75-minute clamping of the uterine circulation. SHAM animals underwent the same procedure except for the clamping. This was followed by full-term vaginal delivery of animals in all groups (FA, SHAM and untreated controls). At 6 months, basal and amphetamine-induced locomotor activity was measured during open field testing. Brain sections were stained for tyrosine hydroxylase (TH) and glial fibrillary acidic protein (GFAP). TH-positive cells and GFAP-positive cells in substantia nigra pars compacta (SN_C) and striatum were counted using design-based stereology. Moreover, TH-immunoreactivity in the striatum was assessed by grey value measurements. Behavioral analysis demonstrated that SHAM and FA showed less basal and amphetamine-induced activity than controls. Histochemically, FA decreased the number of TH-positive neurons in the SN_C and lowered TH-positive in the striatum. Furthermore, more GFAP-positive cells were found in the SN_C and striatum in FA than in either control and SHAM groups. Additionally, FA animals showed ventriculomegaly associated with smaller white matter as well as grey matter volumes. The data show that FA was associated with deficits in both dopamine-related motor behavior and biochemistry. These alterations were associated with nigrostriatal astrogliosis. The present study demonstrates the sensitivity of the dopaminergic system towards FA.     

Ganglioglioma with a tanycytic ependymoma as the glial component

We studied a cystic ganglioglioma (GG) located in the right frontal lobe of the brain. Interestingly, the fibrillary spindle glial cells were often arranged in a fascicular pattern, and the generally uniform, round-to-oval delicate nuclei appeared to resemble those of ependymoma; and the neoplastic neurons often contained neurofibrillary tangles (NFTs). The glial component was positive for glial fibrillary acidic protein and occasionally contained granular or microvesicular structures positive for epithelial membrane antigen. Ultrastructural investigation revealed that the glial cells were ependymal in nature; intracytoplasmic lumina and intercellular microrosettes lined with cilia and microvilli, as well as long zonulae adherentes, were evident. In addition, chromogranin A-positive granular staining, neurosecretory-granule-like structures, and parallel arrays of microtubules were sometimes associated with the blood vessels. We considered the present case to be an unusual example of GG with an ependymoma, more precisely a tanycytic ependymoma, as the glial component; to our knowledge, the existence of ependymoma as the main glial component of this particular tumor has not been described before. The occurrence of NFTs, which has been reported in several cases of GG, was an additional, unusual feature. Received: 3 May 1999 / Revised, accepted: 2 July 1999     

Effects of chronic methamphetamine on the nigral-striatal dopamine system in rat brain: Tyrosine hydroxylase immunochemistry and quantitative light microscopic studies

Chronic administration of methamphetamine (20 mg/kg, IP, every 12 hours for 10 days) produced a large decrease in tyrosine hydroxylase staining axons and terminal boutons in the caudate nucleus in rats when examined 60 days following the final methamphetamine injection. This effect was quantitated using the Leitz Data Acquisition and Display System (DADS) revealing that there was a 74% decrease in tyrosine hydroxylase positive processes in the caudate nucleus. Furthermore, this treatment also produced a large decrease in the number of tyrosine hydroxylase positive staining neuronal perikarya in the pars compacta of the substantia nigra. This effect was also quantitative using the Leitz-(DADS) system, revealing a decrease of 89% in tyrosine hydroxylase positive material. These data demonstrate that chronic administration of methamphetamine produces a long-term loss of tyrosine hydroxylase enzyme in both the cell bodies of the substantia nigra and the nerve terminals in the caudate nucleus. Whether this effect is due to the degeneration of the neurons or some metabolic effect remains to be determined.     

Characterization of the norepinephrine uptake system and the role of norepinephrine in the expression of the adrenergic phenotype by quail neural crest cells in clonal culture

This study investigates the role norepinephrine (NE) may play in regulating the differentiation of quail neural crest cells into sympathoadrenal cells. Cues originating from the embryonic microenvironment are thought to play an important role during development. It is conceivable that NE has a positive regulatory function because adrenergic expression by quail neural crest cells in clonal culture can be inhibited by NE uptake inhibitors such as desipramine (DMI). This possibility is further supported by the notion that in the avian embryo presumptive adrenergic neural crest cells are likely to encounter catecholamines shortly after they have acquired the NE uptake mechanism. Our present data indicate that neural crest cells in clonal culture express a high affinity NE uptake system that can be inhibited by desipramine. As in the embryo, it appears before noticeable levels of catecholamines are accumulated by neural crest cells, as judged by formaldehyde-induced catecholamine fluorescence (FIF). A comparison of the time course of appearance of different adrenergic markers suggests that immunoreactivity against the biosynthetic enzyme tyrosine hydroxylase (TH) may appear first, and that it is followed very closely by the appearance of detectable levels of dopamine-β-hydroxylase (DBH) and the NE uptake mechanism. Accumulation of catecholamines (FIF) is observed last. Addition of exogenous NE leads to an increase in adrenergic expression in vitro as judged by an increase in the number of colonies containing FIF-positive cells as well as cells expressing the biosynthetic enzymes TH and DBH. This suggests that exogenous NE can play a positive regulatory role in the differentiation of quail neural crest cells intossympathoadrenal cells.     

Bisphenol A exposure disrupts the development of the locus coeruleus‐noradrenergic system in mice

It has been reported that bisphenol A (BPA), a widespread xenoestrogen employed in the production of polycarbonate plastics, affects brain development in both humans and rodents. In the present study employing mice, we examined the effects of exposure to BPA (500 μg/kg/day) during fetal and lactational periods on the development of the locus coeruleus (LC) at the age of embryonic day 18 (E18), postnatal 3 weeks (P3W), P8W and P16W. The number of tyrosine hydroxylase‐immunoreactive cells (TH‐IR cells) in females exposed to BPA was decreased, compared with the control females at P3W. At P8W, the number of TH‐IR cells in females exposed to BPA was significantly decreased, compared with the control females, whereas the number of TH‐IR cells in males exposed to BPA was significantly increased, compared with the control males, which resulted in reversed transient sexual differences in the numbers of TH‐IR cells observed in the controls at P8W. However, no significant changes were demonstrated at E18 or P16W. Next, we examined the density of the fibers containing norepinephrine transporter (NET) in the anterior cingulate cortex (ACC) and prefrontal cortex, at P3W, P8W and P16W, because NET would be beneficial in identifying the targets of the LC noradrenergic neurons. There were no significant differences shown in the density of the NET‐positive fibers, between the control and the groups exposed to BPA. These results suggested that BPA might disrupt the development of physiological sexual differences in the LC‐noradrenergic system in mice, although further studies are necessary to clarify the underlying mechanisms.     

Transplantation of human striatal tissue into a rodent model of Huntington''s disease: Phenotypic expression of transplanted neurons and host-to-graft innervation

The present study was undertaken to investigate the phenotypic expression and integration of human striatal neurons transplanted into an animal model of Huntington's disease. Sprague-Dawley rats were anesthetized and subjected to quinolinic acid lesions of the left striatum. Three human fetal cadavers were utilized for transplantation in this study (7,8, and 10 weeks in gestation). The striatal primordia was dissected from each fetus and subsequently dissociated into cell suspensions. Following the initial lesion surgeries (3–4 months), the rats were reanesthetized and transplanted with human striatal cells (400,000 cells per rat). The animals were processed for histochemical analysis 9–17 weeks posttransplantation. Histochemistry was performed utilizing thionin (Nissl staining), acetylcholinesterase, NADPH-diaphorase, and antibodies against tyrosine hydroxylase and glial fibrillary acidic protein. Examination of stained brain sections demonstrate that human striatal transplants grow to fill a substantial portion of the remaining striatum, and contain clusters of immature and mature cells. Acetylcholinesterase activity is present in the transplant neuropil, varying in intensity, and distributed in a heterogeneous fashion. In addition, host afferent dopaminergic fibers penetrate into the transplant, and are occasionally found in patches. NADPH-diaphorase histochemistry revealed medium sized aspiny striatal neurons of donor origin in the transplants. The results of this study are similar to those obtained with rodent fetal striatal transplants, and suggest that human striatal tissue is capable of surviving, expressing normal striatal cell phenotypes, and receiving host dopaminergic innervation.     

Preparation and characterization of a specific antibody for the immunohistochemical detection ofl-DOPA in paraformaldehyde-fixed rodent brains

A rat polyclonal antiserum has been obtained after coupling of L-3,4-dihydroxyphenylalanine (L-DOPA) to larger proteins using a low concentration of glutaraldehyde. The antiserum was tested for its affinity and specificity using an enzyme-linked-immunosorbent-assay (ELISA). From competition experiments, the most immunoreactive compound was found to be the non-reduced L-DOPA conjugate. Our specific L-DOPA antiserum enables us to visualize L-DOPA molecule on brain of guinea pigs and rats. We examined the immunohistochemical distribution of the polyclonal L-DOPA antiserum after the fixation of brains with a mixture of paraformaldehyde and picric acid. The presence of L-DOPA-immunoreactive (IR) neurons and fibers was described in the posterior, dorsal and periventricular hypothalamic areas and in the arcuate nucleus. Finally, the distribution of L-DOPA-IR cells was compared to that of tyrosine hydroxylase (TH)-IR cells, by means of a double staining procedure. The presence of two populations of TH-IR cells (TH-positive/L-DOPA-negative and TH-positive/L-DOPA-positive cells) was described in the dorsal part of the hypothalamus.     

Low‐grade malignant triton tumor in the lumbar spine: A rare variant of malignant peripheral nerve sheath tumor with rhabdomyoblastic differentiation

Malignant peripheral nerve sheath tumor (MPNST) is an uncommon type of sarcoma that arises from peripheral nerve sheaths and rarely involves the spinal roots. The origin of this tumor is thought to be Schwann cells or pluripotent cells of the neural crest. The subgroup of tumors in which malignant Schwann cells coexist with malignant rhabdomyoblasts is termed malignant triton tumor (MTT). MPNSTs can show different degrees of malignancy, but overall spinal MTTs are high‐grade lesions. We report the exceptional instance of a spinal low‐grade MTT in a 39‐year‐old man treated with total surgical removal followed by local radiation therapy. Histological low grade was based on the lack of necrosis, a low grade of atypia, a low mitotic rate and a Ki‐67 labelling index <25%. After 18 months of follow‐up the patient is alive with no evidence of disease. A thorough review of the literature yielded 57 well‐documented spinal MPNSTs. Ten of them corresponded to MTTs, but none showed low‐grade features. An analysis of the clinical, radiological and treatment data was performed to identify factors that might influence the outcome. Overall the 18‐month survival rate was 45% but dropped to 0% in the subgroup of spinal MTTs. Besides, a size exceeding 2 cm, extra‐spinal extension, association with neurofibromatosis and subtotal removal were all related to a worse outcome. In conclusion, spinal MTTs generally exhibit a more aggressive behavior than conventional MPNSTs. The occurrence of a spinal low‐grade MTT with a better prognosis should also be recognized.     

Histological changes in monoaminergic neurons of Long–Evans Cinnamon rats

The Long–Evans Cinnamon rat, an animal model of Wilson’s disease, is an inbred mutant strain with spontaneous hepatitis isolated from Long–Evans rats. The copper concentration in the brains of Long–Evans Cinnamon rats at 4 weeks of age was lower than that of controls, but higher than that of controls at 20 weeks of age. We investigated the tyrosine hydroxylase and 5-hydroxytryptamine immunoreactive fiber densities in the brains of Long–Evans Cinnamon rats aged 4, 10, and 20 weeks by immunohistochemistry, comparing them with Long–Evans Agouti rats used as controls. Tyrosine hydroxylase immunoreactive fiber densities in the cingulate cortex, hippocampus and cerebellum in Long–Evans Cinnamon rats were significantly lower than those of Long–Evans Agouti rats at 4 and 10 weeks of age. On the other hand, 5-hydroxytryptamine immunoreactive fiber densities in the cingulate cortex, caudate-putamen, hypothalamus, and hippocampus in Long–Evans Cinnamon rats were significantly higher than those of controls at 4, 10 and 20 weeks of age. In the cingulate cortex and caudate-putamen, 5-hydroxytryptamine immunoreactive fiber densities became gradually higher with age. The number of aberrant 5-hydroxytryptamine immunoreactive fibers in the cingulate cortex, caudate-putamen, hypothalamus and hippocampus in LEC rats was significantly higher than that of controls. The number of another type of aberrant 5-hydroxytryptamine immunoreactive fibers, which were detected only at 20 weeks of age in the caudate-putamen in LEC rats was significantly higher than that of controls. These results suggest that age-dependent changes in copper concentrations of Long–Evans Cinnamon rats were related to changes in monoaminergic neuron systems.     

The right amygdalar tumor presenting with symptoms of separation anxiety disorder (SAD): a case report

A patient with an astrocytoma of the right-sided amygdala developed symptoms of separation anxiety disorders (SADs). These symptoms significantly subsided after tumor resection. The temporal relationship suggested that the amygdalar tumor could result in the specific symptoms. To our knowledge, this is the first report of SAD as one manifestation of the amygdalar tumor. The tumorigenesis of amygdala resulted in impaired regulation and abnormal activity associated with anticipating anxiety and conditioning. It deserves clinical attention to early detection and intervention.     

Sclerosing fibrous tumor of the cauda equina: a fibroblastic variant of peripheral nerve tumors?

Summary The case of a 43-year-old man with an unusual mesenchymal tumor of the cauda equina is presented. A well-circumscribed firm tumor was found in the lower spinal canal at L1 level. Although a nerve root was involved, the adjacent dura mater or filum terminale was unrelated to the tumor. Microscopically, the tumor was rich in collagen and made up of irregularly intertwining fascicles of fibroblastic spindle cells lacking in nuclear atypia or mitotic activity. Partial broad hyalinization of collagen was another histological feature of the tumor. Histological and immunohistochemical studies failed to reveal any findings that suggested known fibrous neoplasms, such as schwannoma, neurofibroma and meningioma, originating in the nervous system. Ultrastructural features of the tumor cells were consistent with those of fibroblasts. Hence, the present tumor is regarded as a unique pure fibroblastic tumor (fibroma) derived from the interstitium of a nerve root in the cauda equina.