RESUMEN
UNLABELLED: Axoplasmically transported proteins synthesized in neuronal somata labeled by radioactively labeled amino acids (tritium), following local targeted injections for tracing of pathways in the central nervous system using autoradiography. Results from a number of neuronal systems, including: the rat olfactory bulb; cortico-thalamic projections in the mouse; commissural connections of the rat hippocampus; and retinal projections in the monkey and chick are documented. Pathway origins are clear, as the number and distribution of the labeled cells and the normal structure of the injection site is preserved. Light and electron microscopic autoradiography shows that proteins are transported, at two rates: rapid transport (>100mm/day) of fewer proteins accumulating in axon terminals; and, slow transport (1-5mm/day) of the bulk of labeled proteins distributed along the length of axons. Different survival times can be selected to evaluate terminal projection field(s) or pathways from origin to termination. The clarity of autoradiographic labeling of pathways and their terminations is comparable to other techniques (such as the Nauta-Gygax and the Fink-Heimer methods and the electron microscopy of terminal degeneration). Labeled amino acids do not label molecules in fibers of passage and there is no retrograde transport of labeled material from the axon terminals. The functional polarity of fiber pathways can be easily established. We summarize the merits of this technique is based upon an established physiological properties of neurons that are summarized in contrast to currently used techniques dependent upon pathological changes in neurons, axons, or axonal terminals. ARTICLE ABSTRACT: This article considers a heavily cited Brain Research article that reported an extremely important turning point in the ability to demonstrate neuroanatomical pathways in the central nervous system. Using radioactive leucine microinjections into the brain, neurons synthesized proteins from this amino acid that were transported down their axons to the terminal synapses on the target neurons. Tracing the transport of the labeled protein by autoradiography permitted quantitative analysis of projections and pathways. As a result, pathway analysis was transformed from studying the degenerating processes of lesioned neurons to the study of intact pathways in non-manipulated brains. The classical protocol has since been widely applied and used to investigate countless brain circuits. This article is part of a Special Issue entitled SI:50th Anniversary Issue.
Asunto(s)
Autorradiografía/historia , Axones , Encéfalo/anatomía & histología , Técnicas de Trazados de Vías Neuroanatómicas/historia , Neuroanatomía/historia , Neuronas , Animales , Transporte Axonal , Historia del Siglo XX , Ratones , Vías Nerviosas/anatomía & histología , RatasRESUMEN
UNLABELLED: Axoplasmically transported proteins synthesized in neuronal somata labeled by radioactively labeled amino acids (tritium), following local targeted injections for tracing of pathways in the central nervous system using autoradiography. Results from a number of neuronal systems, including: the rat olfactory bulb; cortico-thalamic projections in the mouse; commissural connections of the rat hippocampus; and retinal projections in the monkey and chick are documented. Pathway origins are clear, as the number and distribution of the labeled cells and the normal structure of the injection site is preserved. Light and electron microscopic autoradiography shows that proteins are transported, at two rates: rapid transport (>100mm/day) of fewer proteins accumulating in axon terminals; and, slow transport (1-5mm/day) of the bulk of labeled proteins distributed along the length of axons. Different survival times can be selected to evaluate terminal projection field(s) or pathways from origin to termination. The clarity of autoradiographic labeling of pathways and their terminations is comparable to other techniques (such as the Nauta-Gygax and the Fink-Heimer methods and the electron microscopy of terminal degeneration). Labeled amino acids do not label molecules in fibers of passage and there is no retrograde transport of labeled material from the axon terminals. The functional polarity of fiber pathways can be easily established. We summarize the merits of this technique is based upon an established physiological properties of neurons that are summarized in contrast to currently used techniques dependent upon pathological changes in neurons, axons, or axonal terminals. ABSTRACT: The cytoarchitecture of layer IV in mouse SmI cerebral cortex was examined in.formalin-fixed, Nissl-stained and Cox-fixed, Golgi-Nissl-stained sections cut coronally and tangentially to the pia, A multicellular cytoarchitectonic unit is described in layer IV, roughly cylindrical, 100-400um in diameter, and perpendicular to the pia. Because of their characteristic shape we call these structures barrels. Each barrel is a ring of neurons, the side, which surrounds a less cellular hollow. The nearly acellular reigion surrounding each barrel and separating adjacent barrels is the septum. Barrels are discussed in relation to observations reported in several earlier papers on the mouse cortex. The barrel field (all barrels) has remarkable constancy by all measures: from one hemisphere to the next and from one specimen to the next. A consistent part of the barrel field is the postero-medial barrel subield (PMBSF). Barrels in the PMBSF are larger, elliptical in shape, organized into five distinct rows and their numbers are constant. It is postulated that each barrel in the PMBSF is the cortical correlate of a contralateral mystacial vibrissa (whisker). On the basis of counts of barrels and of all facial sinus hairs a 'one barrel-one vibrissa' hypothesis is proposed. The general hypothesis is that barrels are the morphological manifestation in layer IV of the functional cortical columns discovered by physiologists. The barrels offer excellent opportunities for integrated studies of sensory cerebral cortex at a degree of resolution previously not possible. This article is part of a Special Issue entitled SI:50th Anniversary Issue.
Asunto(s)
Neuroanatomía/historia , Corteza Somatosensorial/anatomía & histología , Animales , Autorradiografía/historia , Historia del Siglo XX , Ratones , Técnicas de Trazados de Vías Neuroanatómicas/historia , Técnicas de Trazados de Vías Neuroanatómicas/métodos , Neuroanatomía/métodos , VibrisasRESUMEN
In situ hybridization is the technique by which specific RNA or DNA molecules are detected in cytological preparations. Basically it involves formation of a hybrid molecule between an endogenous single-stranded RNA or DNA in the cell and a complementary single-stranded RNA or DNA probe. In its original form the probe was labeled with (3)H and the hybrid was detected by autoradiography. The first successful experiments in 1968 involved detection of the highly amplified ribosomal DNA in oocytes of the frog Xenopus, followed soon after by the reiterated "satellite DNA" in mouse and Drosophila chromosomes. Fluorescent probes were developed about ten years later.
Asunto(s)
Autorradiografía/historia , Sondas de ADN/historia , Hibridación in Situ/historia , Cromosomas Politénicos/ultraestructura , Animales , Autorradiografía/instrumentación , Autorradiografía/métodos , ADN/química , ADN/genética , ADN/ultraestructura , Sondas de ADN/síntesis química , Drosophila melanogaster/genética , Colorantes Fluorescentes/síntesis química , Colorantes Fluorescentes/historia , Historia del Siglo XX , Historia del Siglo XXI , Hibridación in Situ/instrumentación , Hibridación in Situ/métodos , Larva/genética , Ratones , Oocitos/metabolismo , Oocitos/ultraestructura , ARN/química , ARN/genética , ARN/ultraestructura , Glándulas Salivales/metabolismo , Glándulas Salivales/ultraestructura , Tritio/química , Xenopus laevis/genéticaAsunto(s)
Citogenética/historia , Cariotipificación , Neoplasias/historia , Patología/historia , Animales , Autorradiografía/historia , Distinciones y Premios , Biopsia con Aguja Fina/historia , Mapeo Cromosómico/historia , Citogenética/normas , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Hungría , Neoplasias/diagnóstico , Neoplasias/virología , Patología/organización & administraciónRESUMEN
The history of receptor autoradiography, its development and applications, testify to the utility of this histochemical technique for localizing radiolabeled hormones and drugs at cellular and subcellular sites of action in intact tissues. Localization of diffusible compounds has been a challenge that was met through the introduction of the "thaw-mount" and "dry-mount" autoradiographic techniques thirty years ago. With this cellular receptor autoradiography, used alone or combined with other histochemical techniques, sites of specific binding and deposition in vivo and in vitro have been characterized. Numerous discoveries, some reviewed in this article, provided information that led to new concepts and opened new areas of research. As an example, in recent years more than fifty target tissues for vitamin D have been specified, challenging the conventional view about the main biological role of vitamin D. The functions of most of these vitamin D target tissues are unrelated to the regulation of systemic calcium homeostasis, but pertain to the (seasonal) regulation of endo- and exocrine secretion, cell proliferation, reproduction, neural, immune and cardiovascular responses, and adaptation to stress. Receptor autoradiography with cellular resolution has become an indispensable tool in drug research and development, since information can be obtained that is difficult or impossible to gain otherwise.