Molecular milestones that signal axonal maturation and the commitment of human spinal cord precursor cells to the neuronal or glial phenotype in development.

Insights into the programmatic induction of neuronal and glial genes during human embryogenesis have depended largely on extrapolations of data derived from experimental mammals. However, the assumptions upon which these extrapolations are based have not been rigorously tested. Indeed, practically no information is available even on the human counterparts of the relatively small subset of well-characterized, developmentally regulated neuron and glial specific genes of the mammalian CNS. Thus, the developmental programs upon which human neural embryogenesis are based remain largely undeciphered. We have addressed this problem in immunohistochemical studies conducted on 22 human fetal spinal cords with gestational ages (GAs) that ranged from 6 to 40 weeks by using monoclonal antibodies to several classes of neuron or glial specific polypeptides. These polypeptides included: representatives of four different types (Types I-IV) of intermediate filament proteins, i.e., vimentin filament protein (VFP), glial fibrillary acidic protein (GFAP), different phospho-isoforms of the high (NF-H), middle (NF-M), and low (NF-L) molecular weight (Mr) neurofilament (NF) subunits, both acidic and basic cytokeratin (CK) proteins; three different microtubule associated proteins (MAPs), i.e., MAP2, MAP5, and tau; two different synaptic or coated vesicle proteins, i.e., synaptophysin (SYP) and clathrin light chain B (LCb); an oligodendroglial specific protein, i.e., myelin basic protein (MBP); and a receptor for a CNS trophic factor, i.e., the nerve growth factor receptor (NGFR).(ABSTRACT TRUNCATED AT 400 WORDS)     

Direct synaptic contacts of catecholamine fibers on neuropeptide Y-containing neurons of the rat cerebral cortex

A direct synapse between catecholamine (CA) fibers and neuropeptide Y (NPY)-containing neurons was demonstrated in rat cerebral cortex by using an immunohistochemical double-staining method under electron microscopy.     

Regeneration through nerve allografts in the cynomolgus monkey (Macaca fascicularis)

With the use of ulnar nerves of cynomolgus monkeys, the present study examined whether basal laminae of Schwann cells can serve as conduits for regenerating axons in nerve allografts from non-human primates. A segment of ulnar nerve was transected distal to the elbow joint one week before grafting. In Group A, a distal segment of the transected nerve was transplanted, after freezing and thawing, into the ulnar nerve of another monkey, at a level that corresponded to that from which the graft was taken. In Group B (the control group), the segment of nerve was grafted in the same manner but without cryotreatment. Two weeks, five weeks, eight weeks, and five months after grafting, the graft and the host nerve were examined with light and electron microscopy. Within two weeks after grafting in Group A, after degradation of the cellular components of the Schwann cells, the basal laminae of the Schwann cells were intact in the form of tubes. Within five weeks, many regenerating axons grew out into these basal lamina tubes in the three-centimeter-long grafts and extended into the host nerve. As seen at the wrist (seven centimeters from the distal suture) five months after grafting, the axons exhibited fully mature myelination both in the graft and in the host nerve. In contrast, in Group B, in which the Schwann cells had not been disrupted by cryotreatment, cellular components and connective-tissue matrices, including basal laminae, had been degraded and had been replaced by invading cells, which filled the endoneurial spaces of the graft. Five months after grafting, axonal growth had been arrested in the graft one centimeter distal to the proximal suture. The beneficial effect in Group A appears to have been the result of the retention and preservation of intact basal laminae of Schwann cells after rapid removal of killed Schwann cells and myelin debris. Killing of Schwann cells by freezing before grafting may abolish the immune response to the Schwann cells in allografts and lead to fragmentation and disruption of myelin, which facilitates the rapid removal of myelin by macrophages.     

Corticotropin-releasing factor-like immunoreactive nerve fibers in the rat superior cervical ganglion and their fine structures

The existence of nerve fibers containing corticotropin-releasing factor (CRF)-like immunoreactivity (CRFI) in the rat superior cervical ganglion (SCG) was demonstrated by using immunocytochemistry. They were found to be extrinsic in origin, because no CRFI neurons were seen in the SCG and decentralization resulted in the disappearance of CRFI fibers in the SCG on the operated side. These findings were also confirmed by immunoelectron microscopic analysis; CRFI fibers contained a number of small clear synaptic vesicles but were devoid of large granular and agranular vesicles. These morphological characteristics are identical to those of the preganglionic fibers. The present immunoelectron microscopic analysis revealed that most of the CRFI fibers in the SCG make synaptic contact predominantly with the dendrites of the principal cells, partly with their somas and rarely with a non-CRFI terminal. Thus, the present study provides direct morphological evidence that CRF directly influences the function of the principal cells of the SCG and that CRFI fibers are preganglionic.     

Demonstration of histaminergic neurons in horizontal cells of guinea pig retina

The existence ofl-histidine decarboxylase (HDC, EC 4.1.1.22)-like immunoreactive (HDC-I) cells in guinea pig retina was demonstrated using antiserum raised against HDC purified from fetal rat liver. The anti-HDC antiserum partially cross-reacted guinea pigl-DOPA decarboxylase (DDC, EC 4.1.1.28), so the histaminergic neurons ware carefully identified. (1) Comparison of HDC-I and DDC-like immunoreactive (DDC-I) cell types in adjacent sections revealed that HDC-I structures were found in some horizontal cells and amacrine cells, and (2) double-staining procedures with anti-HDC antiserum and monoclonal anti-DDC antibody showed that HDC-I horizontal cells had no DDC-I structures, but all the HDC-I amacrine cells had DDC-I structures. From the results, some horizontal cells (with HDC-like immunoreactivities but without DDC-like immunoreactivities) were concluded to be histaminergic.     

Treatment of malignant brain tumors with slowly releasing anticancer drug-polymer composites

The purpose of this study is to present the methodology and results of a clinical trial of local chemotherapy of malignant brain tumors based on slowly-releasing anticancer drug-polymer composites. The slowly releasing drugs were prepared by combining and mutually dispersing anticancer agents with glassified monomers containing 10% polymetacrylic methyl acid and then this compound was frozen at -78 degrees C and exposed to 1 X 10(6) rad of gamma rays from cobalt 60. Thus we prepared a compound of polymers and anticancer agents. We used needle-shaped capsules of this compound. These capsules release the drug very slowly over 40 days. We administered locally to the malignant brain tumors with either slowly releasing mitomycin, slowly releasing adriamycin, slowly releasing ACNU or slowly releasing 5 Fu drugs. The following techniques were employed in implantation these capsules. Implantation into the remaining tumor wall at the time of excision. Implantation into the tumor by CT-guided stereotactic method. We implanted these drugs into tumor of 55 cases, thereafter we conducted both radiation and chemotherapy with ACNU in most patients. This method has the following advantages: It is possible to be employed to different types of anticancer agents. Both dosage and releasing time can be adjusted. It is possible to administer these capsules postoperatively by the stereotactic method. The clinical study consists of 55 patients, 20 cases of anaplastic astrocytoma, 23 cases of glioblastoma multiforme, 5 cases of oligodendroglioma, 3 cases of medulloblastoma and 4 cases of others. Survival rate estimated by Kaplan-Meier method was 47% in glioblastoma at 12 months and 91% in anaplastic astrocytoma at 18 months.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunocytochemical localization of mitochondrial and cytosolic aspartate aminotransferase isozymes in the vestibular end-organs of rats

Summary The localization of mitochondrial (m-) and cytosolic (c-) aspartate aminotransferase (AAT) was examined in the vestibular ganglion neurons and sensory cells in the vestibular end-organs of rats by an indirect immunohistochemical method using antibodies specific for m- and c-AAT. Neurons in the vestibular ganglion were stained by both m- and c-AAT antibodies, but the vestibular sensory cells exhibited only m-AAT-like immunoreactivity and were not labeled by c-AAT. These findings suggested that aspartate is a neurotransmitter in the hair cells of the vestibular end-organs. Offprint requests to: M. Tanaka     

The relationship of the increased level of metallothionein with heavy metal levels in the tissue of the harbor seal (Phoca vitulina)

The relationship between age, heavy metal levels, and a heavy-metal binding protein, metallothionein (MT), in the liver and kidney of the harbor seal (Phoca vitulina) was studied. The cadmium (Cd) level in the liver and the Cd, inorganic mercury (I-Hg), and zinc (Zn) levels in the kidney were increased with body length, suggesting an age-related accumulation of these metals. The MT levels determined by an MT radioimmunoassay showed concentrations of 240 +/- 139 micrograms/g in the liver and 343 +/- 219 micrograms/g in the kidney. These MT levels were shown to be correlated with age. The statistically significant relationship of the MT levels with the levels of Cd and Zn in the liver and those of Cd, Zn, and I-Hg in the kidney suggested that the protein is responsible for the sequestration of these metals as already observed in terrestrial animals.