Biomedical subjects

D L Meinecke

Publications and source records attributed to D L Meinecke.

9 recordsLinked to original sources

PubMed1993

Low-affinity p75 nerve growth factor receptor expression in the embryonic monkey telencephalon: timing and localization in diverse cellular elements.

Monoclonal antibodies against the low-affinity (p75) subunit of the human nerve growth factor receptor have been used to determine the temporal appearance of this receptor and to identify the associated cellular elements in the developing occipital cortex of rhesus monkeys. Adult and fetal brains from embryos at embryonic days 45-121 were used. This embryonic time span includes periods of active neurogenesis, cell migration and initial formation of axonal connections in the cerebral cortex. The first immunolabeling in the developing cerebral wall was seen between embryonic days 56 and 64. The labeling was present in the transient subplate neurons, a small number of axonal processes and pericytes associated with blood vessels. By birth, labeled neurons of the subplate zone disappeared, but immunolabeled axonal processes could now be seen in large numbers in the cortex. These findings are consistent with the role of nerve growth factor in the coordination of cortical differentiation, but not with the initiation of neuronal proliferation, since the emergence of nerve growth factor receptor-labeled elements in the cortex occurs two to three weeks after the onset of neurogenesis in this species. Further, the diverse cellular elements labeled in the fetal cerebrum with the antibodies to the low-affinity nerve growth factor receptor suggests that a receptor or receptors associated with growth factor signaling for more than one growth factor family are recognized by these antibodies. Differential timing in the expression of families of growth factor receptors may be one mechanism by which developing neurons in the cerebral cortex could respond to the different signals which guide such processes as synaptogenesis and morphogenesis.

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PubMed1992

Expression of GABA and GABAA receptors by neurons of the subplate zone in developing primate occipital cortex: evidence for transient local circuits.

Several lines of evidence suggest that the transient subplate zone of the embryonic mammalian telencephalon could influence cortical development through synaptic or trophic interactions with growing cortical afferents and migrating neurons. Since such interactions may involve neurotransmitters and their receptor molecules, we have examined the expression of GABA and subunits of the GABAA/benzodiazepine receptor complex in the occipital lobe of embryonic rhesus monkeys by immunochemistry and in situ hybridization. We found that during the second half of gestation, when the subplate zone reaches peak maturity in this species, many neurons can be immunolabeled with both GABA antisera and monoclonal antibodies against GABAA receptor subunits. The most robust labeling occurs at approximately embryonic day (E)125 (birth is at E165). Electron microscopic observations of receptor subunit-immunolabeled material confirmed that subunits of the GABAA receptor are localized in the subplate neurons and their dendritic processes. In many instances the reaction product is associated with the plasma membranes of labeled processes, some of which form symmetrical synapses with small unlabeled axon terminals. The results of in situ hybridization are in accord with the results of receptor subunit immunochemistry. From E80 to E141, hybridization signal for GABAA receptor subunit mRNA occurs in the subplate zone and increases steadily to peak levels between E125 and E141. The present results reveal that all the elements necessary for the formation of functional GABAergic synaptic circuitry are present in the subplate zone. Further, the ages showing the most pronounced receptor and transmitter expression in this primate coincide with the ingrowth of major cortical afferent systems. Taken together, these findings suggest that GABAergic local neuronal circuits in the subplate may be involved in the development of long tract connections stationed in this zone prior to their transfer to the overlying cortical plate.

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PubMed1990

Developmental expression of GABA and subunits of the GABAA receptor complex in an inhibitory synaptic circuit in the rat cerebellum.

The temporal relationship between the expression of a transmitter and its corresponding receptor may provide important insights into the development of synaptic circuits in the central nervous system. Here we examined the emergence of the inhibitory transmitter GABA, and subunits of the GABAA/benzodiazepine receptor complex in a well-characterized cerebellar circuit formed by granule cells and the synapses they make with Golgi II neurons in the cerebella of rats ranging in age from birth to 21 days. The presence of GABA was determined immunocytochemically. The presence of the GABAA receptor was demonstrated by localizing the alpha 1 subunit of the receptor using in situ hybridization and immunochemical localization of a 50 kDa benzodiazepine-binding subunit using monoclonal antibodies. Germinal cells of the external granular layer which give rise to granule cells did not express the GABAA receptor at any age. Similarly, receptor labeling could not be detected in granule cells during their postmitotic migratory period. In the internal granular layer, immature postmigratory granule cells are unlabeled. The expression of GABAA receptor subunits was first observed on the fifth postnatal day (P5) and then only in the more mature granule cells which have well elaborated dendrites in contact with presynaptic elements. The number of labeled neurons increased over the subsequent ages examined. Presynaptic elements in association with the dendrites of labeled granule cells had ultrastructural features characteristic of Golgi II cell axon terminals. These elements demonstrate GABA transmitter as early as P3, preceded by 2-3 days receptor labeling in the granular layer. Therefore, granule cells express GABAA receptor subunits only after they have completed migration and their dendrites have become involved in specific synaptic circuits, including innervation by GABAergic afferents.

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PubMed1989

GABAA/benzodiazepine receptor-like immunoreactivity in rat and monkey cerebellum.

The cellular and subcellular localization of GABAA/benzodiazepine receptor-like immunoreactivity in the rat and monkey cerebellum has been studied with a monoclonal antibody (E9) directed against the alpha-subunit of purified GABAA/benzodiazepine receptors. At both the light and electron microscopic level E9 immunoreactivity is located in all 3 layers of the cerebellar cortex and within the deep cerebellar nuclei. The reaction product accumulates within the cytoplasm of neurons and their dendrites but axons are not immunoreactive. Glial cells in the white matter and the cortical layers are also unlabeled, although in some instances Bergmann glia do contain reaction product. The overall distribution and cellular and subcellular localization of E9 immunoreactivity is identical for both monkey and rat cerebellum. On the basis of cell size, morphology, and location it is evident that E9 immunoreactivity occurs in examples of all 5 neuronal types in the cerebellar cortex: Purkinje cells, Golgi type II cells, granule cells, and stellate and basket cells. However, the distribution of the reaction product within the cells is more selective. For example, electron microscopy demonstrates that axonal processes and terminals are not E9 immunoreactive with the single exception of the mossy fiber terminals in the granular layer. Also, examples of unlabeled axon terminals resembling those derived from Golgi type II cells, basket cells, and stellate cells form synapses with immunoreactive dendrites and cell bodies in the cortical layers. Finally, in the deep cerebellar nuclei unreactive axon terminals make symmetric synapses with immunostained neurons and dendrites. These results show that E9 monoclonal antibodies label neurons and portions of their processes which are postsynaptic in GABA-mediated inhibitory circuits, and demonstrates that this antiserum can be used as a morphological marker for cells which make GABAA/benzodiazepine receptors.

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PubMed1987

GABA immunoreactive neurons in rat visual cortex.

An antiserum to gamma-aminobutyric acid (GABA) was used in a light and electron microscopic immunocytochemical study to determine the morphology and distribution of GABA-containing neurons in the rat visual cortex and to ascertain whether all classes of nonpyramidal neurons in this cortex are GABAergic. The visual cortex used for light microscopy was prepared in such a way that the antibody penetrated completely through tissue sections, and in these sections large numbers of GABA immunoreactive neurons were apparent. The labeled neurons could be identified as being either multipolar, bitufted, bipolar, or horizontal neurons. In layers II through VIa, GABA immunostained cells were distributed uniformly and accounted for approximately 15% of all neurons, but in layer I all neurons appeared to be immunostained. Electron microscopy of GABA immunostained visual cortex prepared to ensure good fine structural preservation confirmed the presence in layers II through VIa of numerous immunoreactive bipolar neurons, both small and large varieties, as well as multipolar and bitufted neurons. Additionally, electron microscopy reveals that astrocytes are frequently GABA immunoreactive. From a correlated light and electron microscopic evaluation of neurons in GABA immunostained visual cortex, it was possible to confirm which kinds of neurons are GABAergic and what proportion of the neuronal population they represent. Thus, from an analysis of some 950 neurons, it was found that pyramidal neurons were never immunoreactive and that except for 20% of the bipolar cell population, all examples of other types of nonpyramidal neurons encountered in this material were GABA immunoreactive.

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PubMed1987

Vasoactive intestinal polypeptide immunoreactive neurons in the primary visual cortex of the cat.

When cat visual cortex (area 17) is reacted with an antibody to vasoactive intestinal polypeptide (VIP) a variety of neuronal types is labelled. Many of the labelled neurons are bipolar in form and are most common in layers II and III, although significant numbers of bipolar neurons are also encountered in layer V. Multipolar cells are also labelled. These are most frequent in layer IV and have a variety of shapes. In layer I, the labelled cells are of three varieties, i.e. horizontal bipolar cells, horizontal bitufted cells and multipolar neurons, while in layer VI the few VIP-positive neurons are horizontal bipolar cells. This suggests that all of the VIP-labelled neurons in cat area 17 are non-pyramidal in form, and this has been confirmed by electron microscopy. In these preparations, axon terminals are also labelled and under the light microscope it can be seen that these terminals occur both within the neuropil and around the cell bodies of some neurons, particularly neurons in layers II and III. Electron microscopy has shown that all of the labelled axon terminals form symmetric synapses and that those in the neuropil synapse with the shafts of smooth dendrites. These axodendritic synapses account for about 90% of the synapses formed by the labelled axon terminals. The remainder of the labelled axon terminals synapse with the cell bodies of pyramidal neurons. Parallels are drawn between these results and those previously obtained by examining those neuronal elements labelled with VIP antibodies in rat visual cortex.

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PubMed1986

Somatostatin immunoreactive neurons in rat visual cortex: a light and electron microscopic study.

Somatostatin immunoreactive neurons in rat visual cortex were examined in the light and electron microscopes using an antibody to the tetradecapeptide form of somatostatin. Somatostatin immunoreactive neurons were found to belong only to non-pyramidal classes. They are of five main types: multipolar neurons with either thin or thick dendrites; small and large bipolar neurons; bitufted neurons; horizontal neurons; and neurons in the subcortical white matter. Of the immunoreactive neurons, multipolar neurons are the most common and account for 30% of the population, while bipolar and bitufted neurons make up 25% and 15% of the immunoreactive population, respectively; the least common somatostatin immunoreactive neurons are the horizontal and subcortical white matter neurons. Occasional multipolar neurons with thick dendrites have a prominent ascending dendrite so that they resemble pyramidal cells in the light microscope, but electron microscopic examination confirms that, like all other somatostatin-positive cells, they are non-pyramidal neurons, for they have both symmetric and asymmetric synapses on their cell bodies. Somatostatin-positive neurons are distributed among all the cortical layers and the subcortical white matter but they are more common in two laminae, one coinciding with layer II/III and the other with layers V and VI. The multipolar and bipolar neurons are distributed in similar proportions in these upper and lower cortical laminae, while bitufted neurons are more common in upper laminae and horizontal neurons are predominantly located in layer VI.

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PubMed1984

Fine structure of dividing astroglia and oligodendroglia during myelin formation in the developing mouse spinal cord.

To study the morphology and cellular relationships of dividing glial cells during myelin formation, were perfused newborn and 5-day mouse pups and embedded slices of cervical, thoracic, and lumbar cord for light and electron microscopic study. In semithin epon sections stained with toluidine blue, all levels of spinal cord at both ages contained mitotic glia in gray columns and funiculi. In electron micrographs of funiculi, dividing astroglia containing bundles of glial filaments, many glycogen granules, and had large processes extending into the surrounding neuropil. Cytoplasmic organelles of many immature interphase oligodendroglia and mitotic oligodendroblasts were similar and included microtubules, clusters of free ribosomes, and scattered profiles of granular endoplasmic reticulum. Unlike astroglia, dividing oligodendroblasts lacked large processes and in metaphase they were ellipsoids and had smooth plasma membranes. When these cells were studied in alternating serial thin and semithin sections over 10-15 micrometers distances, we did not identify connections between myelin sheaths and mitotic oligodendroblasts. Our findings indicate that oligodendroglia in developing white matter multiply before developing large processes. Our data also suggest that oligodendroglia do not divide while forming myelin.

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PubMedDate not supplied

Early expression of GABA-containing neurons in the prefrontal and visual cortices of rhesus monkeys.

Light and electron microscopic immunohistochemistry was used to examine the time of emergence and distribution of GABA-containing cells in an association (prefrontal) and primary sensory (visual) region of the telencephalon at progressive fetal and postnatal stages of cortical maturation in the rhesus monkey. Thirty fetuses and six postnatal monkeys were examined beginning at embryonic day 41 (E41), the start of cortical neurogenesis, to birth (E165) and proceeding to maturity (greater than 5 years of age). The emergence and major developmental modifications in the distribution of immunoreactive neurons in both areas examined were nearly identical. GABA-immunolabeled neurons were present throughout the full thickness of the cerebral wall, including the cortical plate and the developmentally transient marginal, subplate, and ventricular zones, as early as E41. An important and surprising result was that a subset of bipolar migrating neurons in the subplate zone also contained GABA at these early stages. GABA-containing neurons in the ventricular and subventricular zones disappeared after E100, when neurogenesis is completed. In contrast, the number of immunoreactive multipolar and bipolar neurons within the subplate zone diminished between E100 and E131. By the first postnatal week, the distribution and density of GABA-containing neurons in the cortex appeared qualitatively similar to that observed in mature monkeys. The early appearance of GABA in cortical neurons and its expression by a population of migrating neurons suggest that a subset of cortical neurons may be committed to a transmitter phenotype independent of synaptic interactions and prior to attaining their adult positions in the maturing cerebral cortex.

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