Life-threatening hemophagocytic syndrome triggered by disseminated toxoplasmosis in a young patient with previously unknown AIDS.

Hemophagocytic syndrome is a rare life-threatening disorder that can be triggered by various conditions such as HIV infection and opportunistic agents. We report a case of disseminated toxoplasmosis complicated with severe hemophagocytic syndrome and revealing an unknown acquired immunodeficiency syndrome. The patient presented with multiple organ failure in intensive care unit. Once diagnosed, he benefitted from etoposide infusion, administration of specific anti-toxoplasmosis treatments and secondary antiretroviral therapy. He was alive at intensive care unit discharge and returned home with little sequalae. This case illustrates both the importance of rapid investigations of hemophagocytic syndrome etiologies in HIV positive patients and the necessity to prompt etoposide and specific treatments in order to improve potentially dramatic outcomes.

Glutamic acid decarboxylase isoform 65 immunoreactivity in the motor thalamus of humans and monkeys: γ-aminobutyric acidergic connections and nuclear delineations.

The neurotransmitter γ-aminobutyric acid (GABA) plays an important role in the motor thalamic nuclei. This report analyzes the distribution of the GABA-producing enzyme glutamic acid decarboxylase isoform 65 (GAD65), stained with monoclonal antibody, in human and rhesus monkey thalami and compares it with staining patterns of some widely used cytoskeletal and calcium binding protein markers. GAD65 immunoreactivity distinctly labeled two systems: fibers and terminals of basal ganglia thalamic afferents and local circuit neurons, revealing fine features of GABAergic circuitry in the human thalamus. Gross distribution patterns of GAD65 were identical in human and rhesus monkey thalami. The area displaying specific staining of large-caliber beaded fibers coincided with nigro- and pallidothalamic afferent territories previously identified in monkeys with anterograde tracers. Accordingly, a similarly stained region in the human thalamus was considered basal ganglia territory. Except for cytoarchitecture, no specific markers differentiating between the nigro- and pallidothalamic projection zones within this territory were found. GAD65 staining in the cerebellar afferent territory reflected organization of its local circuit neuron network, distinguishing it from adjacent nuclei. Specific GAD65 staining pattern and negative calcium binding protein immunoreactivity identify the cerebellar afferent territory in humans. It is subdivided further into ventral and dorsal regions based on the cytoskeletal protein SMI31 staining pattern. The nuclear outlines revised according to the results are compared with those of Hassler (Schaltenbrand G and Bailey P [1959] Einfuhrung in die stereotaktishen Operationen mit einem Atlas des menschlichen Gehirns, vol 3. Stuttgart: Thieme) and discussed in light of the ongoing controversy regarding delineations of the motor thalamic nuclei in humans.

Ontogenic study of the influence of tissue plasminogen activator (t-PA) in neonatal excitotoxic brain insult and the subsequent microglia/macrophage activation.

Intracerebral injections of ibotenic acid in neonatal mice produced white and gray matter lesions that mimic some aspects of the acquired cerebral injuries observed in human newborns (i.e. periventricular leukomalacias in preterm newborns and post-ischemic cortical necrosis in at term infants). We have evaluated the effects of tissue plasminogen activator inactivation (t-PA-/-) on the effects of ibotenic acid (0.01-20 microg), and on F4/80 labeling of microglia/macrophages at different stages. Three ontogenic periods have been identified. In mice injected the day of birth, postnatal (P) day 0, ibotenic acid induced neuronal migration disorders together with low local microglial activation in wild-type and t-PA-/- mice. In P2 and P5 mice, ibotenic acid induced diffuse microglial activation in the whole cortex and subcortical areas; e.g. caudate nucleus and septum. In wild-type mice, cystic lesions of the white matter were consistently observed, surrounded by macrophages. In t-PA-/- mice, noncystic lesions filled of macrophages were more frequent than cysts. Macrophages were virtually absent in the gray matter. White and gray matter lesions were reduced in t-PA-/- mice. The plasmin inhibitor aprotinin reduced white and gray matter lesions only in wild-type mice injected with high ibotenic acid doses (2.5-5 microg). During this period, a transient F4/80 immunoreactive cell population was detected in the cingulum. At P10, the salient lesion characteristic was a large gray matter lesion containing macrophage accumulation. Microglial activation was confined to the injection site in the white matter. t-PA-/- mice showed reduced lesion size under high doses (>5 microg) of ibotenic acid. Similarly, aprotinin diminished the lesion in wild-type animals exposed to 10 microg ibotenic acid. These data demonstrate that t-PA and microglia do not actively participate in the migration disorders induced in P0 mice. Conversely, t-PA was implicated in cyst formation in older (P2-P10) mice, and in their subsequent growth. t-PA was also involved in GM lesions, probably through an inflammatory process involving macrophages.

Early microglial activation following neonatal excitotoxic brain damage in mice: a potential target for neuroprotection.

Previous studies in a mouse model of neonatal excitotoxic brain damage mimicking the brain lesions in human cerebral palsy showed microglial activation within 24 h after intracerebral injection of the glutamatergic analog ibotenate. Using this model, we studied the expression of CD-45 antigen, a marker of blood-derived cells, by these activated microglial cells labeled by Griffonia simplicifolia I isolectin B4. Immunohistochemistry performed during early development of excitotoxic lesions showed that most cells labeled with the isolectin B4 were CD-45-negative, suggesting that these early activated microglial cells were deriving chiefly from resident microglia and not from circulating monocytes. We also directly tested the hypothesis that activated resident microglia and/or blood-derived monocytes play a role in the pathophysiology of excitotoxic brain damage. Repeated i.p. administrations of chloroquine, chloroquine+colchicine, minocycline, or an anti-MAC1 antibody coupled to the toxin saporin before and/or after ibotenate injection induced a significant reduction in the density of isolectin B4-positive cells. This inhibition of resident microglial and/or blood-derived monocytes activation was accompanied by a significant reduction in the severity of ibotenate-induced brain lesions (up to 79% lesion size reduction with the highest minocycline dose) as well as of ibotenate-induced cortical caspase-3 activation (49% reduction).

Confirmation of the retinopetal/centrifugal nature of the tyrosine hydroxylase-immunoreactive fibers of the retina and optic nerve in the weaver mouse.

The number of tyrosine hydroxylase-immunoreactive fibers in the nerve fiber layer is increased in the retina of the weaver compared to control mice (Dev. Brain Res. 121 (2000) 113). To confirm the retinopetal/centrifugal nature of these fibers, a newly devised whole-mounted optic nerve technique allowed us to determine, during development, their first appearance within the optic nerve (post-natal day 12) compared to retina (post-natal day 13). One such fiber was also observed looping in the retina of a monkey fetus.

Intrauterine growth retardation does not alter the distribution of tyrosine hydroxylase-immunoreactive neurons of A8, A9 and A10 groups in the rat: a three-dimensional reconstruction study.

The purpose of the present investigation was to evaluate possible effects of severe prenatal hypotrophy on the number and spatial distribution of tyrosine hydroxylase-immunoreactive neurons of the A8, A9 and A10 cell groups in the rat brain. Prenatal hypotrophy was induced in rat pups by ligaturing one uterine artery in pregnant rats on the 17th day of gestation. This procedure induces a severe growth retardation, which is never caught up with, even at adulthood. In both control and growth-retarded adult rats, serial coronal sections were cut through the retrorubral field, the substantia nigra, and the ventral tegmental area (A8, A9 and A10 cell groups, respectively). The number of tyrosine hydroxylase-immunoreactive neurons was determined, and their spatial localization was recorded by means of a 3-dimensional reconstruction software developed in our laboratory. Our 3-dimensional models provide a visual illustration of the heterogeneous continuum formed by the dopaminergic neurons. They illustrate the difficulty in demarcating the A8, A9 and A10 cell groups. Finally, our results show that intrauterine growth retardation did not affect either the number or the 3-dimensional organization of tyrosine hydroxylase-immunoreactive neurons in the adult rat brain.

Structure of longitudinal brain zones that provide the origin for the substantia nigra and ventral tegmental area in human embryos, as revealed by cytoarchitecture and tyrosine hydroxylase, calretinin, calbindin, and GABA immunoreactions.

In a previous work, mapping early tyrosine hydroxylase (TH) expressing primordia in human embryos, the tegmental origin of the substantia nigra (SN) and ventral tegmental area (VTA) was located across several neuromeric domains: prosomeres 1-3, midbrain, and isthmus (Puelles and Verney, [1998] J. Comp. Neurol. 394:283-308). The present study examines in detail the architecture of the neural wall along this tegmental continuum in 6-7 week human embryos, to better define the development of the SN and VTA. TH-immunoreactive (TH-IR) structures were mapped relative to longitudinal subdivisions (floor plate, basal plate, alar plate), as well as to radially superposed strata of the neural wall (periventricular, intermediate, and superficial strata). These morphologic entities were delineated at each relevant segmental level by using Nissl-stained sections and immunocytochemical mapping of calbindin, calretinin, and GABA in adjacent sagittal or frontal sections. A numerous and varied neuronal population originates in the floor plate area, and some of its derivatives become related through lateral tangential migration with other neuronal populations born in distinct medial and lateral portions of the basal plate and in a transition zone at the border with the alar plate. Some structural differences characterize each segmental domain within this common schema. The TH-IR neuroblasts arise predominantly within the ventricular zone of the floor plate and, more sparsely, within the adjacent medial part of the basal plate. They first migrate radially from the ventricular zone to the pia and then apparently move laterally and slightly rostralward, crossing the superficial stratum of the basal plate. Several GABA-IR cell populations are present in this region. One of them, which might represent the anlage of the SN pars reticulata, is generated in the lateral part of the basal plate.

Expression of calbindin D28K in the dopaminergic mesotelencephalic system in embryonic and fetal human brain.

A subset of tyrosine-hydroxylase (TH) neurons of the substantia nigra (A9) containing calbindin D28K (CaBP) appeared to be less vulnerable to cell death induced by Parkinson's disease than the subset containing dopamine (DA) alone. Because grafting procedures of fetal human neurons are increasingly used in the therapy of Parkinson's disease, it is important to study the development of DA neurons coexpressing CaBP. In humans, the genesis of TH immunoreactivity of A9, of the ventral tegmental area (A10), and of the retrorubral area (A8) occurred during a 2-week period from the 4. 5th gestational week (g.w.) in the ventricular zone of the floor plate and the contiguous basal plate of the mesencephalon and diencephalon, i.e., the prosomeres p1-p3. Double-immunolabeled TH-CaBP neurons were detected from 5.5 g.w. on, in the first wave of DA neuron's migration, and were observed in their final residence in the dorsal A9 by 10.5 g.w. Calretinin immunoreactivity was expressed in TH-immunoreactive (IR) neurons from 10.5 g.w. on. Ascending TH-CaBP-IR axons were observed toward the telencephalon from 6-7 g.w. , reaching the anlage of the nucleus accumbens and amygdaloid complex at 10.5 g.w., but were not detected in the ganglionic eminence at this latter stage. Dopaminergic patches were detected at 13 g.w. in the anlage of the putamen, but no TH-CaBP-IR fibers were observed in the matrix at this stage. In conclusion, even if CaBP immunoreactivity was detected in TH-IR cell bodies during the embryonic period, the TH-CaBP-IR axonal terminal was observed earlier in some limbic-related areas than in the matrix compartment of the basal ganglia in humans.

Defect of tyrosine hydroxylase-immunoreactive neurons in the brains of mice lacking the transcription factor Pax6.

In the CNS, the lack of the transcription factor Pax6 has been associated with early defects in cell proliferation, cell specification, and axonal pathfinding of discrete neuronal populations. In this study, we show that Pax6 is expressed in discrete catecholaminergic neuronal populations of the developing ventral thalamus, hypothalamus, and telencephalon. In mice lacking Pax6, these catecholaminergic populations develop abnormally: those in the telencephalon are reduced in cell number or absent, whereas those in the ventral thalamus and hypothalamus are greatly displaced and densely packed. Catecholaminergic neurons of the substantia nigra (SN) and the ventral tegmental area (VTA) do not express Pax6 protein. Nevertheless, mice lacking Pax6 display an altered pathfinding of SN-VTA projections: instead of following the route of the medial forebrain bundle ventrally, most of the SN-VTA projections are deflected dorsorostrally at the pretectal-dorsal thalamic transition zone and in the dorsal thalamic alar plate. Moreover, some catecholaminergic neurons are displaced dorsally to an ectopic location at the pretectal-dorsal thalamic transition zone. Interestingly, from the pretectal-dorsal thalamic to the dorsal thalamic-ventral thalamic transition zones, mice lacking Pax6 display an ectopic ventral to dorsal expansion of the chemorepellant/chemoattractive molecule, Netrin-1. This may be responsible for both the altered pathway of catecholaminergic fibers and the ectopic location of catecholaminergic neurons in this region.

Paradoxical increase of tyrosine hydroxylase-immunoreactive retinopetal fibers in the weaver mouse.

Weaver mice undergo apoptosis of the granule cell precursors of the cerebellum and nonapoptotic death of mesencephalic dopaminergic cells during post-natal development. In contrast, the number of retinal dopaminergic cells was transiently increased in weaver compared to control mice [C. Savy, E. Martin-Martinelli, A. Simon, C. Duyckaerts, C. Verney, C. Adelbrecht, R. Raisman-Vozari, J. Nguyen-Legros, Altered development of dopaminergic cells in the retina of weaver mice, J. Comp. Neurol. 1999;412:656-668]. While re-examining the retinas, we observed, in the nerve fiber layer, retinopetal tyrosine hydroxylase-immunoreactive fibers, which were dramatically increased in number throughout development and adulthood in the weaver compared to control mice.

Independent controls for neocortical neuron production and histogenetic cell death.

We estimated the proportion of cells eliminated by histogenetic cell death during the first 2 postnatal weeks in areas 1, 3 and 40 of the mouse parietal neocortex. For each layer and for the subcortical white matter in each neocortical area, the number of dying cells per mm(2) was calculated and the proportionate cell death for each day of the 2-week interval was estimated. The data show that cell death proceeds essentially uniformly across the neocortical areas and layers and that it does not follow either the spatiotemporal gradient of cell cycle progression in the pseudostratified ventricular epithelium of the cerebral wall, the source of neocortical neurons, or the 'inside-out' neocortical neuronogenetic sequence. Therefore, we infer that the control mechanisms of neocortical histogenetic cell death are independent of mechanisms controlling neuronogenesis or neuronal migration but may be associated with the ingrowth, expansion and a system-wide matching of neuronal connectivity.

Altered development of dopaminergic cells in the retina of weaver mice.

Postnatal degeneration of dopaminergic (DA) cells is known to occur in mesencephalic nuclei of mutant weaver mice, whereas retinal DA content is reported to be unchanged in the adult animal. To determine whether morphological changes occur in the weaver retinal DA system, we compared weaver and control developing and adult retinas after tyrosine hydroxylase (TH) immunohistochemistry. The density and distribution of DA cells were analyzed using Dirichlet tessellation. Not only was no DA cell loss found in adult weaver retinas, but we even observed an increase in DA cells in weaver compared to control retinas between postnatal days 14 and 30. Furthermore, some unusual features were found during the latter period: atypical cells (representing a maximum of 12% of the whole DA cell population) were observed, and these differed from typical DA cells in terms of both location (slightly more external within the inner nuclear layer) and appearance (flat somata, round and clear nuclei, thick dendritic trunks emerging laterally and giving rise to horizontal processes). Some of the atypical cells were intermingled in a delicate network lying in a more outer focal plane than the main DA plexus. The expression of GIRK2, a G protein-related inward rectifying K(+) channel responsible for the weaver syndrome, was investigated. Although no GIRK2 labeling was demonstrated in DA cells, its possible involvement in the transient disturbances observed in the weaver DA retinal system is discussed.

Distribution of the catecholaminergic neurons in the central nervous system of human embryos and fetuses.

The catecholaminergic cell groups in the human brain, denominated from A1 to A17, display some striking anatomical differences with those described in the rodent. These differences are essentially observed in the extent of the dopaminergic neurons and especially their axonal fields in the telencephalon. Immunocytochemistry for tyrosine-hydroxylase and dopamine-ss-hydroxylase allowed the visualization of the precocious human catecholaminergic groups as early as 4.5 postovulatory weeks. Maps of tyrosine-hydroxylase positive neurons generated in the different rhombomeres, midbrain, and prosomeres are shown following the prosomeric model introduced by Puelles and Rubenstein [(1993) Trends Neurosci. 16:472-476]. Such a description is convenient to compare catecholaminergic systems in different mammalian species and provide clear anatomical landmarks of the embryonic substantia nigra (midbrain and prosomeres 1 and 2), that are necessary for transplantation of neural tissue in Parkinson's disease. The development and early specification of the dopaminergic neurons expressing calbindin D28K phenotype in the substantia nigra and in the ventral tegmental area are described. The catecholaminergic axons enter the anlage of the cerebral cortex just after the formation of the cortical plate, from 7 postovulatory weeks on. They invade the subplate layer where they wait for 4 weeks before penetrating the cortical plate. At midgestation, the different areas and layers of the frontal cerebral wall are invaded by the catecholaminergic axons, before the layering of the cortex is completed, in a pattern of fiber distribution similar to that described in the adult human brain. The early pattern of development of the catecholamine systems appeared to be phylogenetically well preserved in mammals, but specific features emerging during the differentiation period are unique to humans.

Anosmin-1 is a regionally restricted component of basement membranes and interstitial matrices during organogenesis: implications for the developmental anomalies of X chromosome-linked Kallmann syndrome.

Kallmann syndrome is a developmental disease characterized by gonadotropin-releasing hormone (GnRH) deficiency and olfactory bulb hypoplasia. The gene underlying the X chromosome-linked form, KAL-1, has been identified for several years, yet the pathogenesis of the disease is not understood. By immunohistofluorescence and immunoelectron microscopy, we establish that the KAL-1 encoded protein, anosmin-1, is a transient and regionally restricted component of extracellular matrices during organogenesis in man. Anosmin-1 was detected in the basement membranes and/or interstitial matrices of various structures including bronchial tubes, mesonephric tubules and duct, branches of the ureteric bud, muscular walls of the digestive tract and larger blood vessels, precartilaginous models of skeletal pieces, muscle tendons, head mesenchymes, inner ear, and forebrain subregions. Our results suggest that this protein acts as a local, rather than a long-range, cue during organogenesis. In the olfactory system, anosmin-1 was detected from week 5 onward. The protein was restricted to the olfactory bulb presumptive region and later, to the primitive olfactory bulbs. We therefore suggest that the genetic defect underlying X-linked Kallmann syndrome disrupts the terminal navigation of the early olfactory axons or directly affects the initial steps of olfactory bulb differentiation. The mechanism of the GnRH deficiency is also discussed, relying on the evidence that anosmin-1 is present in the medial walls of the primitive cerebral hemispheres, along the rostro-caudal migratory pathway of the GnRH-synthesizing neurons, at 6 weeks. Finally, the present results strongly suggest that the renal aplasia observed in about one third of the affected individuals results from primary failure of the collecting duct system.

Early neuromeric distribution of tyrosine-hydroxylase-immunoreactive neurons in human embryos.

A segmental mapping of brain tyrosine-hydroxylase-immunoreactive (TH-IR) neurons in human embryos between 4.5 and 6 weeks of gestation locates with novel precision the dorsoventral and anteroposterior topography of the catecholamine-synthetizing primordia relative to neuromeric units. The data support the following conclusions. (1) All transverse sectors of the brain (prosomeres in the forebrain, midbrain, rhombomeres in the hindbrain, spinal cord) produce TH-IR neuronal populations. (2) Each segment shows peculiarities in its contribution to the catecholamine system, but there are some overall regularities, which reflect that some TH-IR populations develop similarly in different segments. (3) Dorsoventral topology of the TH-IR neurons indicates that at least four separate longitudinal zones (in the floor and basal plates and twice in the alar plate) found across most segments are capable of producing the TH-IR phenotype. (4) Basal plate TH-IR neurons tend to migrate intrasegmentally to a ventrolateral superficial position, although some remain periventricular; those in the brainstem are related to motoneurons of the oculomotor and branchiomotor nuclei. (5) Some alar TH-IR populations migrate superficially within the segmental boundaries. (6) Most catecholaminergic anatomical entities are formed as fusions of smaller segmental components, each of which show similar histogenetic patterns. A nomenclature is proposed that partly adheres to previous terminology but introduces the distinction of embryologically different cell populations and unifies longitudinally analogous entities. Such a model, as presented in the present study, is convenient for resolving problems of homology of the catecholamine system across the diversity of vertebrate forms.

Comigration of tyrosine hydroxylase- and gonadotropin-releasing hormone-immunoreactive neurons in the nasal area of human embryos.

Tyrosine hydroxylase (TH) immunoreactive (IR) central catecholaminergic neurons have been observed in human CNS from 4.5 gestational weeks (g.w.) on [Verney, C., Zecevic, N. and Puelles, L. Eur. J. Neurosci., Suppl. 8 (1995) 7044; Zecevic, N. and Verney, C., J. Comp, Neurol., 351 (1995) 509-535]. We describe here a discrete TH-IR cell population localized in the rostral nasal region during embryonic development. Tyrosine hydroxylase-IR cells spread from the olfactory placode towards the basal and medial telencephalon. They follow the same migration path as the gonadotropin-releasing hormone (GnRH)-IR hypothalamic neurons. Tyrosine hydroxylase-IR neurons are first detected at 4.5 g.w., while GnRH-IR cells are visualized later at 5.5 g.w. Double immunocytochemical labeling reveals the presence of three neuronal populations comigrating along the developing vomeronasal-nervus terminalis complex. These populations express either one or both TH and GnRH phenotypes depending on their position in the migration route. At 6 g.w., most of the neurons express TH immunoreactivity as they leave the vomeronasal organ whereas most of the GnRH-IR neurons are detected closer to the CNS and in the CNS itself. These results emphasize the early phenotypic heterogeneity of the different migrating neuronal populations generated in the olfactory placode in humans. At later stages, very few TH-IR neurons are detected in the anterior forebrain suggesting a transient expression of TH immunoreactivity within these neuronal populations.

Early postnatal changes of the dopaminergic mesencephalic neurons in the weaver mutant mouse.

In adult weaver (wv) mutant mice up to 70% of the mesostriatal dopaminergic neurons are lost and major alterations of the dopaminergic dendrites of the substantia nigra have been described. We sought to determine the time of onset of these alterations. Cell counts of the main dopaminergic (DA) mesencephalic cell groups (A8, A9, A10), as labeled with tyrosine hydroxylase immunocytochemistry were done in wild-type and homozygous wv/wv pups. No loss of the DA neurons, was detectable at postnatal day 7 (P7), while reductions in substantia nigra (and retrorubral area) amounted to 35% at P14 and 47% by P21. On the other hand, the severe reduction of dopaminergic dendrites, particularly of their distal compartments was already visible from P3 on. During the first postnatal week, this was associated to abnormal clustering of the dopaminergic neurons. These early neuritic alterations were present, though to a milder degree, in heterozygous (wv/+) mice.

Cajal-Retzius neurons in human cerebral cortex at midgestation show immunoreactivity for neurofilament and calcium-binding proteins.

Along with subplate neurons, Cajal-Retzius cells (CRc) are the first neurons to be generated in the cortical anlage. Studies of their chemical content, such as neurofilament and calcium-binding proteins, might give indications on their role in cortical development at midgestation in human fetuses (20-24 gestation weeks), when the CRc are morphologically mature. Cajal-Retzius cells were immunolabeled with antibodies to subunits of neurofilament proteins SMI31 and SMI32. The SMI32 antibodies (directed against the nonphosphorylated epitope) specifically labeled the CR cell bodies, dendrites, and proximal axons in a Golgilike fashion. Specific acetylcholinesterase activity is known to be typical of all the CRc, and double labeling for SMI32 immunoreactivity and acetylcholinesterase histochemistry demonstrated that all the CRc exhibited SMI32 immunoreactivity. The SMI31 antibodies (directed against the phosphorylated epitope) exclusively labeled the CRc axons, forming a dense positive network in the deep one-half of layer 1. This plexus was much denser than the one described with the Golgi method (Marin-Padilla, 1990: J. Comp Neurol 239:89-105). Calbindin D28k, parvalbumin, and calretinin immunoreactivities were visualized in the CRc. Double-labeling experiments showed that most of the CRc contained both calbindin and calretinin and sometimes parvalbumin. These colocalizations revealed a chemical heterogeneity within the CRc population even though they were described as morphologically homogeneous. These colocalizations of calcium-binding proteins in the CRc differed from the other nonpyramidal cortical neurons where calbindin, calretinin, and parvalbumin are contained in different (mutually exclusive) neuronal populations. Based on the morphological features and differential chemical contents described for the CRc, different hypotheses on their possible role and fate are discussed.