1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neuroblastic apoptosis in the subventricular zone is caused by 1-methy-4-phenylpiridinium (MPP(+)) converted from MPTP through MAO-B.

Intraperitoneal 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration induces apoptosis of subventricular zone (SVZ) doublecortin (Dcx)-positive neural progenitor cells (migrating neuroblasts, A cells). Actually, a metabolite of MPTP, 1-methy-4-phenylpiridinium (MPP(+)), is responsible for neural progenitor cell toxicity. In the present study, to examine whether the MPTP-induced SVZ cell apoptosis is caused directly by MPP(+) metabolized through monoamine oxidase B (MAO-B), MPTP or MPP(+) was intracerebroventricularly (icv) injected into C57BL/6 mice. At Day 1 postinjection, many terminal deoxynucleotidyl transferase-mediated dUTP endlabeling (TUNEL)-positive cells were observed in the SVZ of both low (36 µg) and high (162 µg) dose MPTP- and MPP(+)-injected mice. The number of Dcx-positive A cells showed a significant decrease following high dose of MPTP- or MPP(+)-injection on Days 1 and 3, respectively, whereas that of EGFR-positive C cells showed no change in mice with any treatment. In addition, prior icv injection of a MAO-B inhibitor, R(-)-deprenyl (deprenyl), inhibited MPTP-induced apoptosis, but not MPP(+)-induced apoptosis. MAO-B- and GFAP-double positive cells were detected in the ependyma and SVZ in all mice. It is revealed from these results that icv injection of MPTP induces apoptosis of neural progenitor cells (A cells) in the SVZ via MPP(+) toxicity. In addition, it is suggested that the conversion from MPTP to MPP(+) is caused mainly by MAO-B located in ependymal cells and GFAP-positive cells in the SVZ.

Anti-allodynic effect of intracerebroventricularly administered antioxidant and free radical scavenger in a mouse model of orofacial pain.

AIMS: To evaluate possible effects of the intracerebroventricular (icv) injection of either O-Tricyclo [5.2.1.0(2,6)] dec-9-yl dithiocarbonate potassium salt (D609), a potent antioxidant and inhibitor of phosphatidylcholine specific phospholipase C (PtdCho-PLC) and acid sphingomyelinase (ASMase), or the spin trap/free radical scavenger N-tert-Butyl-alpha-phenylnitrone (PBN), on mechanical allodynia induced by facial carrageenan injection in mice. METHODS: Balb/c mice received icy injection of D609/PBN plus facial carrageenan injection, and the number of face wash strokes to von Frey hair mechanical stimulation of the maxillary skin was quantified. PtdCho-PLC and ASMase activities were also assayed in the brainstem, thalamus, and somatosensory cortex. RESULTS: Mice that received the icy injection of 10 nmol D609 plus facial carrageenan injection showed significantly fewer face wash strokes evoked by von Frey hair stimulation (indicating reduced mechanical allodynia) at 1 and 3 days post-injection, compared to mice that received icy injection of isotonic saline plus facial carrageenan injection. Mice that received icy injection of 1.13 micromol PBN plus facial carrageenan injection likewise showed significantly fewer face wash strokes after facial carrageenan injection, compared to isotonic saline-injected plus carrageenan-injected controls. D609 injection also resulted in significantly reduced ASMase activity in the brainstem, thalamus, and somatosensory cortex 3 days after injection, compared to controls. CONCLUSION: The icv injections of D609 and PBN were effective in reducing mechanical allodynia after facial carrageenan injection-induced pain. Together, the results point to a possible role of central nervous system sphingolipids and/or free radicals in orofacial pain.

Succinic dehydrogenase histochemistry reveals the location of the putative primary visual and auditory areas within the dorsal ventricular ridge of Sphenodon punctatus.

In turtles, crocodilians, lizards and snakes, the dorsal ventricular ridge (DVR) is a nuclear cell mass that contains distinct visual and auditory thalamorecipient cell groups. In the tuatara (Sphenodon punctatus), the DVR is not organized into diverse cell groups but instead possesses a trilaminar cytoarchitecture resembling that characteristic of the telencephalic cortex in reptiles. To determine if visual and auditory fields might also be present in the DVR of Sphenodon punctatus, we used succinic dehydrogenase (SDH) histochemistry, which has been shown to delineate the visual and auditory fields of the DVR in turtles, crocodilians and lizards. We also used acetylcholinesterase (AChE) histochemistry to determine the boundary between the DVR and the basal ganglia in Sphenodon. We found an SDH-rich region in the neuropil ventral to the cell plate of the rostrolateral DVR and a slightly less intense SDH-rich zone in the neuropil deep to the cell plate of the ventromedial DVR. These SDH-rich zones appear to be located at the apical dendrites of the neurons of the adjacent cell plate. These SDH-rich zones were clearly located within the DVR and were distinct from the AChE-rich striatal part of the basal ganglia, which occupied the ventrolateral wall of the telencephalon. Based on findings in other reptiles, it seems likely that the SDH-rich zone in rostrolateral DVR represents the zone of termination of nucleus rotundus visual input to the DVR, whereas the zone in ventromedial DVR represents the zone of termination of nucleus reuniens auditory input. Because a trilaminar DVR such as that in Sphenodon might be the primitive DVR condition for reptiles, our results suggest that the cytoarchitecture of the DVR and the synaptic organization of its thalamic sensory input in the common ancestor of living reptiles might have been much like of the dorsal cortex.

Immunocytochemical localization of the catecholamine-synthesizing enzymes, tyrosine hydroxylase and dopamine-beta-hydroxylase, in the hypothalamus of cattle.

Immunocytochemical staining for the presence of catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine beta-hydroxylase, was used to characterize the regional distribution of catecholaminergic neurons in the hypothalamus and adjacent areas of domestic cattle, Bos taurus. In steers, heifers and cows, tyrosine hydroxylase-immunoreactive perikarya was located throughout periventricular regions of the third cerebral ventricle, in both anterior and retrochiasmatic divisions of the supraoptic nucleus, suprachiasmatic nucleus, and ventral and dorsolateral regions of the paraventricular nucleus, dorsal hypothalamus, ventrolateral aspects of the arcuate nucleus, along the ventral hypothalamic surface between the median eminence and optic tract, and in the posterior hypothalamus. Immunostained perikarya ranged from small (10-20 microns, parvicellular) to large (30-50 microns, magnocellular) and were of multiple shapes: round, triangular, fusiform or multipolar, often with 2-5 processes of branched arborization. There were no dopamine-beta-hydroxylase immunoreactive perikarya observed within the hypothalamus and adjacent structures. However, both tyrosine hydroxylase and dopamine-beta-hydroxylase immunoreactive fibers and punctate varicosities were observed throughout regions of tyrosine hydroxylase immunoreactivity perikarya. Generally, the location and pattern of hypothalamic tyrosine hydroxylase immunoreactivity and dopamine-beta-hydroxylase immunoreactive were similar to those reported for most other large brain mammalian species, however, there were several differences with commonly used small laboratory animals. These included intense tyrosine hydroxylase immunoreactivity of perikarya within the retrochiasmatic division of the supraoptic nucleus (ventral A15 region), the absence of tyrosine hydroxylase immunoreactive perikarya below the anterior commissure or within the bed nucleus of stria terminalis (absence of the dorsal A15 region), an abundance of tyrosine hydroxylase immunoreactive perikarya within the ependymal layer of the median eminence, heavy innervation of the arcuate nucleus with dopamine-beta-hydroxylase immunoreactive fibers and varicosities, and the paucity of dopamine-beta-hydroxylase immunoreactive throughout the median eminence.

Primary culture of circumventricular organs from the rat brain lamina terminalis.

A primary culture system of cells derived from two circumventricular organs (CVO) of the rat brain was established. The subfornical organ (SFO) and the organum vasculosum of the lamina terminalis (OVLT) were dissected from the rostral wall of the third ventricle and its cells taken into culture after mechanical dissociation. The cells were cultured in a modified microculture chamber system ensuring relatively high cell density despite their low absolute number. When animals were injected with Evans blue prior to cell preparation, the macroscopically visible penetration of the dye into the parenchyma of the CVOs could be used as guidance during tissue isolation and labelled cells could be identified in culture. Cultured CVO neurones and astrocytes were identified using antibodies against cell type specific marker proteins. The histochemical NADPH-diaphorase staining was used for the detection of nitric oxide synthase in tissue sections of both CVOs and in their cultured neurones. In addition, angiotensin II (ANG II)-evoked elevations of the intracellular Ca2+ concentration ([Ca2+]i) in single cultured OVLT neurones were measured. The described methods will be useful for further characterization of CVO neurones and astrocytes.

Angiotensin-converting enzyme is present in the subfornical organ and other circumventricular organs of the rat.

Angiotensin-converting enzyme (ACE, kininase II, EC 3.4.15.1) activity was measured by a radiochemical assay in isolated circumventricular organs of the rat. ACE activity was unevenly distributed, with a 100-fold difference between the lowest (subcommisural) and the highest (subfornical organ) activities. Our results suggest that angiotensin II could be locally formed in circumventricular organs and especially in the subfornical organ. The high angiotensin-converting enzyme activity in the subfornical organ could indicate a physiological role of endogenous angiotensin II in this structure.

The influence of electrical stimulation of certain brain regions on the concentration of acetylcholinesterase in rabbit cerebrospinal fluid.

Electrical stimulation of either the caudate nucleus, substantia nigra or lateral hypothalamus led to an increase in the concentration of acetylcholinesterase in cerebrospinal fluid collected from the cisterna magna of the rabbit. This effect was not due to gross cell damage, fluid loss or contamination with blood plasma. Cerebrospinal fluid collected from the lateral ventricle contained a lower concentration of acetylcholinesterase than that collected from the cisterna magna; furthermore, stimulation of the ipsilateral caudate nucleus caused no change in the concentration of this enzyme in ventricular cerebrospinal fluid. After placing a large lesion in the ipsilateral substantia nigra, the 'resting' concentration of acetylcholinesterase in cisternal cerebrospinal fluid was 43% of the value in unoperated rabbits and stimulation of the caudate nucleus no longer led to an increase in the concentration of acetylcholinesterase in cisternal cerebrospinal fluid. The possibility is discussed that when the caudate nucleus is stimulated, acetylcholinesterase is released from the substantia nigra.

Vascular and brain dopamine beta-hydroxylase activity in young spontaneously hypertensive rats.

Dopamine beta-hydroxylase activity was higher in mesenteric vessels, adrenal glands, and serum of 3-week-old spontaneously hypertensive rats but lower in the locus coeruleus than it was in the control Wistar-Kyoto rats. The results support the concept that the nervous system is an important regulator of blood pressure.

Monoamine oxidase localization in the ependyma and infundibular recess in the catfish Clarias batrachus and its probable significance.

The presence of monoamine oxidase (MAO) in the cerebrospinal fluid (CSF) and MAO positive tracts bridging the CSF and the subependyma strongly suggest the involvement of CSF in the neuroendocrine control of hypophysial function.