Synaptic development in long‐term organized cultures of murine hypothalamus

Masurovsky, E. B.; Benitez, Helena H.; Murray, Margaret R.

doi:10.1002/cne.901430302

Cited by 36 publications

(6 citation statements)

References 30 publications

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“…Honi and Shinohara (1979) demonstrated in newborn rats that the firing rate of warm sensitive units increased during the first 20 days in postnatal rats and the warm sensitive units were detected earlier in the neonatal stage than the cold sensitive one. According to the morphological studies of hypothalamic neurons both in vivo (Reier et al 1977) and in organ culture (Masurovsky et al 1971), increase in number of synapses and advances in degree of maturity are evident in hypothalamic neurons for about one month. The development of the warm and cold sensitivities might be maturation.…”

Section: Discussionmentioning

confidence: 99%

“…The method of organ culture of the hypothalamus was described in our previous report (Nakayama et al, 1978). The procedure to get the subdivisional hypothalamic tissue was described by Masurovsky et al (1971). This subdivisional hypothalamic tissue was dissected into 4 coronal sections about 300-500 um thickness from rostrum to cauda and the residual hypothalamic tissue was discarded.…”

Section: Methodsmentioning

confidence: 99%

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Temperature sensitivity of the preoptic and anterior hypothalamic neurons in organ culture.

Hori

Nakayama

1982

Tohoku J. Exp. Med.

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Single unit activities were recorded from the preoptic and anterior hypothalamic (POAH) explants obtained from newborn mice brain to study neural temperature sensitivities in the period of 10-34 days culture. The firing rate of recorded units was less than 13 impulses per sec at 35°C. Fifteen warm sensitive, 2 cold sensitive and 37 thermally insensitive units were obtained. The temperature sensitivities of the cultured POAH neurons were similar to those of in vivo neurons, i.e. the discharge frequencies changed smoothly in proportion to 1-5°C changes in local temperature.The thermal responses of warm sensitive units increased after 18 days and decreased over 30 days in vitro. Two warm sensitive neurons were identified and observed under the optical microscope. The cold sensitive units were unable to be recorded until about 20 days in vitro. It became evident that the neural temperature sensitivities of POAH neurons were maintained in an absence of the extrahypothalamic afferents, temperature-sensitive neurons; hypothalamus; organ culture; temperature regulation

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Temperature sensitivity of the preoptic and anterior hypothalamic neurons in organ culture.

Hori

Nakayama

1982

Tohoku J. Exp. Med.

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“…Media from substantia nigra and locus ceruleus cultures contained substantial quantities of DA. Addition of reserine (10 pM) to the medium depleted locus ceruleus neurons of th amines.…”

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confidence: 98%

“…The controlled conditions of the culture situation could yield data on the metabolism, pharmacological responsiveness, receptor specificity, and developmental characteristics of defined, homogeneous populations of neurons. To date, neuronal culture systems of mammalian brain have utilized entire brain (1, 2), individual cortical (3-6) or lower mesencephalic (7)(8)(9)(10)(11) The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.…”

mentioning

confidence: 99%

“…The controlled conditions of the culture situation could yield data on the metabolism, pharmacological responsiveness, receptor specificity, and developmental characteristics of defined, homogeneous populations of neurons. To date, neuronal culture systems of mammalian brain have utilized entire brain (1, 2), individual cortical (3-6) or lower mesencephalic (7)(8)(9)(10)(11) Norepinephrine/Dopamine Determinations. NE and DA levels were measured by a modified radioenzymatic method (17,18) based on reaction of catecholamines with partially purified catechol-O-methyltransferase (S-adenosyl-L-methionine:catechol O-methyltransferase, EC 2.1.1.6), using Sadenosyl[3H]methionine as the methyl donor (19).…”

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confidence: 99%

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Explant cultures of catecholamine-containing neurons from rat brain: Biochemical, histofluorescence, and electron microscopic studies

Schlumpf

Shoemaker

1977

Proc. Natl. Acad. Sci. U.S.A.

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ABSTRACr Norepinephrine (NE~producing cells of the nucleus locus ceruleus and dopamine (DA)producing cells of the substantia nigra were dissected microscopically from embryonic rat brain, explanted, and maintained in cu ture for up to 5 weeks. The cultured neurons of both brain regions showed normal maturation of axons and dendrites and formed ultrastructurali defined synaptic contacts. Fluorescence microscopy of cultured neurons from both brain regions showed typical in situ cytological features: long axonal processes with multiple varicosities for locus ceruleus cultures, and smooth, wispy nonvaricose processes in the substantia nigra cultures. All cultures processed for fluorescence microscopy contained specific catecholamine-fluorescent cells. BY radioenzyme assay for The long survival time in culture of locus ceruleus cells, the normal appearance of fluorescent cell bodies and processes, the apparent development of morphologically specialized interneuronal connections, and the ability to synthesize and store NE make these cultures ideally suited for neurophysiological recording as well as morphological, biochemical, and pharmacological experiments.Many advantages would accrue from the ability to culture specific neuronal nuclei from mammalian brain. The controlled conditions of the culture situation could yield data on the metabolism, pharmacological responsiveness, receptor specificity, and developmental characteristics of defined, homogeneous populations of neurons. To date, neuronal culture systems of mammalian brain have utilized entire brain (1, 2), individual cortical (3-6) or lower mesencephalic (7)(8)(9)(10)(11) The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Cell survival and architectonic differentiation of the hypothalamic mamillary region of the newborn mouse in culture,

Sobkowicz

Bleier

Monzain

1974

J of Comparative Neurology

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The development of the hypothalamic mamillary region of the newborn mouse has been studied in 519 cultures. The nuclear groups of the mamillary configuration as well as the infundibular nucleus may survive for at least 50 days. The surviving neuronal groups undergo in culture a further cytological and structural organization. The presence of ependymal tissue at the surface of the substrate enhances ependymal-glial proliferation and this in turn supports the survival and new growth of the nerve cells in the adjacent areas. On the other hand, degeneration of white matter, especially if it occurs between the substrate and the overlying neuronal groups, has a highly detrimental effect leading to a profound regional neuronal loss. The survival of neuronal groups, the new axonal growth and the preservation of the cellular architectonics, which make possible their identification in culture, indicate that the mamillary complex has a capacity for regeneration during its development in isolation. The marnillary region and the infundibular nucleus display different developmental patterns and different structural organizations of their cells and axons. As a result, the culture seems to be divided morphologically into two distinctly separate regions.The hypothalamic area has been a subject of extensive studies for many years. Morphological studies have been supplemented by experiments in which varying degrees of isolation from the remaining neural or endocrine centers have been produced i n a n experimental animal. At least some results seem to be constant; namely, a transneuronal degeneration of the medial mamillary nuclei after destruction of the limbic cortex i n a newborn or young animal (Gudden, 1880; Cowan and Powell, '55; Bleier, '69), a severe retrograde degeneration of both medial and lateral mamillary nuclei after anterior thalamic injury or severance of their axons (Powell and Cowan, '54; Fry et al., '63; Fry and Cowan, ' 7 2 ) , and a preservation of premamillary and infundibular nuclei regardless of the degree of isolation (Bleier et al., '66) newborn animal, at a time when chances for the survival of any nerve cell after axonal injury are expected to be minimal and those for the survival of the medial mamillary neurons are known to be nil? The tissue culture method seemed to be ideal for such a n investigation. The use of this method in this study brought an additional challenge to the project. As far as it is known, all studied regions of the fetal, newborn or young developing nervous system including the hypothalamus (Murray, '65; Lumsden, '68; Masurovsky et al., '71) show a capacity for survival, new growth and some differential organization during development in culture. While it is known that the paraventricular and supraoptic 355

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Synaptic development in long‐term organized cultures of murine hypothalamus

Cited by 36 publications

References 30 publications

Temperature sensitivity of the preoptic and anterior hypothalamic neurons in organ culture.

Temperature sensitivity of the preoptic and anterior hypothalamic neurons in organ culture.

Explant cultures of catecholamine-containing neurons from rat brain: Biochemical, histofluorescence, and electron microscopic studies

Cell survival and architectonic differentiation of the hypothalamic mamillary region of the newborn mouse in culture,

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