Search PubMed⌕ Search

Biomedical subjects

H Hatanaka

Publications and source records attributed to H Hatanaka.

At least 217 records · Page 12Linked to original sources

Nerve-growth-factor-dependent and cell-density-independent survival of septal cholinergic neurons in culture from postnatal rats.

We have established a primary neuronal cell culture technique from the postnatal (P11 to P15) rat CNS to study the nerve growth factor (NGF) response to basal forebrain cholinergic neurons. The survival of septal cholinergic neurons in culture was monitored both by the determination of choline acetyltransferase activity and by counting acetylcholinesterase-positive cells. Cells obtained from postnatal septal regions were found to require a plentiful oxygen supply during the dissociation of the cells. NGF-mediated survival of the septal cholinergic neurons was similarly observed in the cultures by using different plating cell densities up to 12.5 X 10(5) cells/cm2. These results suggest that the promotion by NGF of cell survival in culture is independent of plating cell density.

Acetylcholinesterase↗

Interstitial deletion of long arm of chromosome 2(q31q33).

We describe the case of a 4-month-old girl with interstitial deletion of the long arm of chromosome 2(46,XX,del(2) (q31q33]. Clinical features included intrauterine growth retardation, psychomotor delay, antimongoloid slanting of the palpebral fissures, hypertelorism, low set ears, cleft palate, micrognathia, luxatio coxae and pes varus. It is suggested that the gene for soluble isocitrate dehydrogenase (IDH1) is located on 2q33.3. The activity of serum IDH1 was in the normal range in this patient.

Abnormalities, Multiple↗

Synthesis and secretion of human nerve growth factor by Saccharomyces cerevisiae.

The DNA coding for human nerve growth factor (hNGF) was chemically synthesized and introduced into Saccharomyces cerevisiae. Expression and secretion of hNGF was obtained by use of the yeast phosphoglycerate kinase-encoding gene promoter and the pre-pro sequence of the yeast alpha-mating factor. Immunoblotting with antiserum raised against a protein A-hNGF fusion protein, allowed the detection of an immunoreactive material secreted into the culture medium. A preparation from the culture medium, partially purified by ion-exchange column chromatography, stimulated neurite outgrowth from rat pheochromocytoma PC12h cells.

Amino Acid Sequence↗

Increase in choline acetyltransferase activity in septum of rats after transient forebrain ischemia: a possible role of factors released in the hippocampus.

Choline acetyltransferase (ChAT) activity increased in rat septum 2 weeks after a transient forebrain ischemia. Extracts were prepared from hippocampus in which CA1 pyramidal neurons had been selectively destroyed by the ischemic insult. ChAT activity in septal neuronal cultures treated with these extracts for 6 days was significantly higher than that in control cultures.

Animals↗

Interleukin-6 as a neurotrophic factor for promoting the survival of cultured basal forebrain cholinergic neurons from postnatal rats.

Human recombinant interleukin-6 (IL-6, B-cell stimulating factor-2) was capable of supporting neuronal survival in cholinergic neuron culture, prepared from 10-day-old rat brain septal region. Cell survival of the cultured cholinergic neurons was estimated by measuring the remaining choline acetyltransferase (ChAT) activities after 6 days of culture. IL-6 at a concentration of 5 ng/ml maintained a more than 3-fold higher ChAT activity in the culture as compared with that in cultures without IL-6. The maximal dose of IL-6 was near 50 ng/ml. The concomitant addition of mouse nerve growth factor (NGF) and IL-6, both at maximal doses, had a synergistic effect on cholinergic cell survival. These results indicate that IL-6 can act as a neurotrophic agent, independent of the action of NGF, supporting neuronal survival of cultured postnatal rat septal cholinergic neurons. On the other hand, IL-6 did not affect the differentiation of the cultured embryonic rat septal cholinergic neurons, differently from the differentiation action by NGF.

Animals↗

Transplantation of septal cholinergic neurons to the hippocampus improves memory impairments of spatial learning in rats treated with AF64A.

Embryonic septal neurons were transplanted into damaged hippocampus in adult rats which had received lateral ventricular administration of AF64A, a cholinergic neurotoxin. About 3 months after transplantation, the rats with bilateral septal grafts showed significant improvement in the radial maze and T-maze tasks. Many ingrowths of acetylcholinesterase (AChE)-positive fibers originating from the grafts were observed in the hippocampus of the rats which showed good performance in these learning tasks. These results indicate that transplantation of septal cholinergic neurons into the AF64A-treated hippocampus may induce at least partial recovery in learning tasks believed to involve the hippocampus.

Animals↗

Recovery of hippocampal cholinergic activity by transplantation of septal neurons in AF64A treated rats.

Embryonic septal neurons were transplanted into the hippocampus of adult rats which had received lateral-ventricular administration of AF64A, a cholinergic neurotoxin, and the effects on hippocampal cholinergic activity were studied. One week after AF64A administration, we injected dissociated septal cell suspension into the dorsal hippocampus, unilaterally. About 3 months after the transplantation, acetylcholine (ACh)-rich septal grafts formed extensive acetylcholinesterase (AChE)-positive fibers into the host hippocampus, recovering choline acetyltransferase (ChAT) level only in the grafted side. These results indicate that septal implants can produce a partial recovery of the cholinergic activity in the chemically damaged hippocampus.

Acetylcholinesterase↗

Nerve growth factor promotes survival of retrogradely labeled hippocampus-projecting neurons in the rat basal forebrain in vitro.

The effect of nerve growth factor (NGF) on the survival of neurons projecting to the hippocampus from developing medial septum and vertical limb of the diagonal band was studied in vitro. The neurons had previously been labeled retrogradely in vivo with fluorescent latex microspheres. The microspheres were injected bilaterally into the hippocampus of 5-day-old rats. Twenty to 24 h after the injection, cells from the basal forebrain were dissociated with papain and cultured for 3-5 days. The number of microsphere-labeled neurons in the culture with supplementation of NGF was much greater than that without NGF. The result clearly indicates a survival-promoting effect of NGF on these projection neurons.

Animals↗

IGF-II-like immunoreactivity in human tissues, neuroendocrine tumors, and PC12 cells.

We looked for cells that reacted with monoclonal antibodies against rat insulin-like growth factor II (IGF-II). We found them in human fetal kidneys in the 14th week of gestation and in adult adrenal medullas but not in human liver, pancreas, pituitary gland, or thymus. Every pheochromocytoma in a group of 20 had IGF-II-like immunoreactive cells, as did three out of four ganglioneuroblastomas (one of which was heterotransplantable in athymic nude mice) and one carotid body tumor. Using the electron microscope we localized the immunoreactivity in pheochromocytoma cells to their neurosecretory granules of the non-catecholamine type. We failed to find any IGF-II-reactive cells among the pancreatic islet cell tumors, neuroblastomas, medullary carcinomas of the thyroid, retinoblastomas, and Wilms' tumor we studied. One line of human neuroblastoma cells did show some immunoreactivity after treatment with dbcAMP. Additionally, the rat pheochromocytoma cell line PC12 and its subline PC12h occasionally produced a few immunostained cells. These results strongly indicate that human IGF-II is primarily produced in paraganglionic tissues and tumors.

Adrenal Gland Neoplasms↗

Intracellular storage and evoked release of acetylcholine from postnatal rat basal forebrain cholinergic neurons in culture with nerve growth factor.

Cholinergic neurons from the septum area, the vertical limb of the diagonal band of Broca, and the nucleus basalis of Meynert of postnatal 13-day-old rats were cultured with or without nerve growth factor (NGF) conditions. Total choline acetyltransferase (ChAT) activities, acetylcholine (ACh) contents, and survival numbers of cholinergic neurons in culture from each of three distinct regions were increased by NGF treatment, but little difference was found in cellular ChAT activities and ACh contents obtained in cultures with or without NGF. The result shows that NGF promotes the survival of cholinergic neurons from 13-day-old rats. Furthermore, the release of ACh from cultured neurons was investigated. The cells cultured with NGF showed a larger increase of the high K+-evoked ACh release than those cultured without NGF. However, NGF had no effect on spontaneous release. This suggests that NGF could regenerate and sustain the stimulation-evoked release mechanisms of ACh in cultured cholinergic neurons from postnatal rats.

Acetylcholine↗

Skin blood flow in diabetic patients during cold loading.

We measured periflux blood flow (PBF) in people's fingers and its response to cold loading with a laser Doppler flowmeter. As an index in the cold loading test we used cold recovery time (CRT). Among normal people, CRT increased with age. Diabetics had significantly longer CRTs than did age-matched controls. CRT seems to be useful for evaluating the skin microcirculation in diabetics and the aged.

Adolescent↗

Transplantation of fetal and early postnatal rat septal cholinergic neurons cultured in serum-free and serum-containing medium with nerve growth factor.

Fetal rat (E17-E19) septal neurons were cultured in a defined, serum-free medium for 6-8 days with or without nerve growth factor (NGF) and transplanted into the hippocampus or the surrounding ventricle of 28 adult rats denervated of its septal input by a fimbria-fornix transection. The cholinergic septal neurons, which were visualized by acetylcholinesterase (AChE) histochemistry, always survived in transplantation to the adult brains from nearly pure neuronal cultures. Although choline acetyltransferase (ChAT) activity of septal neurons in culture was greatly increased (5.59-fold) by the addition of NGF to the defined medium, this ChAT induction appeared to have little effect on the subsequent survival or growth of the septal neurons after transplantation. These results demonstrate that survival of cultured fetal septal cholinergic neurons following transplantation is not dependent upon the presence of NGF or serum- or glia-derived factors during the preliminary culture. Postnatal rat (P4) septal neurons cultured for 5 days in serum-containing medium with NGF were also successfully transplanted in one of 3 cases.

Acetylcholinesterase↗