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A E Hendrickson

Publications and source records attributed to A E Hendrickson.

At least 19 recordsLinked to original sources
PubMed1996

Immunohistochemical characterization of developing and mature primate retinal blood vessels.

PURPOSE: To characterize developing retinal blood vessels with vascular markers and to relate the histochemical profile of maturing vessels to morphologic stages in retinal vascular development. METHODS: Vessels were examined in frozen and paraffin-embedded retinas and in wholemounts of Macaca monkeys ranging in age from fetal day 75 (F75) to adulthood. Endothelial cells were visualized immunohistochemically using antisera to von Willebrand's factor and CD31 with lectins Ulex europaeus, Bandeiraea simplicifolia, peanut agglutinin, Ricinis communis, and wheat germ agglutinin, and by ATPase and ADPase enzymatic histochemistry. Antibodies to vascular basement membrane and matrix markers laminin, fibronectin, and collagen types I and VIII, and antisera recognizing cell cycle-specific nuclear proteins (cyclin, Ki-67, Mib-1) also were used. RESULTS: Newly formed and mature vessels were reactive with reagents specific for CD31, von Willebrand's factor, types I and VIII collagens, laminin, fibronectin, U. europaeus, R. communis, and peanut agglutinin. Wheat germ agglutinin labeled vessels only after pretreatment with neuraminidase. All vascular markers appeared simultaneously, but some were distributed differentially between capillaries and larger vessels, along the central-peripheral extent of a vascular plexus, and among different vascular laminae. Markers of vessels failed to label spindle-shaped presumed vascular precursor cells lying peripheral to the advancing vessels during development. Spindle cells exhibited cyclin, Ki-67, and Mib-1 immunoreactivity. CONCLUSIONS: Immature and mature vitread and sclerad vessels displayed histochemical profiles that were qualitatively similar but that had subtle quantitative differences. Results do not support identification of spindle-shaped cells as vascular precursors in the developing monkey retina and are discussed in relation to mechanisms of retinal vascularization.

Aging↗
PubMed1996

Primary culture and characterization of microvascular endothelial cells from Macaca monkey retina.

PURPOSE: To develop methods for the culture of microvascular endothelial cells (EC) from Macaca monkey retina and to investigate their propagation and survival in vitro. METHODS: Endothelial cells from capillary fragments were cultured on fibronectin-coated dishes in QB-58 serum-free medium containing 20 microliters/ml bovine retinal extract, 90 micrograms/ml heparin, 10% fetal bovine serum, and 10% monkey serum. Non-EC were removed manually. Endothelial cell-specific properties were assessed by endocytosis of acetylated low-density lipoprotein (ac-LDL) and by immunocytochemical staining. The response to growth factors was assayed by 3H-thymidine incorporation. The synthesis of matrix macromolecules was studied by metabolic labeling with 3H-proline and identification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis-immunoblotting. RESULTS: Under these culture conditions, migrating cells emerged from capillary fragments after 1 to 2 days and formed large colonies by 1 week. Cells exhibited a mean doubling time of 44.5 hours during the first 3 to 5 days of culture and 23 hours at 6 to 8 days in culture, and they formed a confluent monolayer by 12 to 14 days. These cells demonstrated uptake of ac-LDL, expressed von Willebrand factor and the cell adhesion protein CD31, and did not contain smooth muscle alpha-actin. Before purification, 92% of the cells in primary cultures were identified as EC. The EC could be maintained in vitro for more than 1 month without the addition of growth factors; however, basic fibroblast growth factor and vascular endothelial growth factor each stimulated cell replication. Secreted extracellular proteins included fibronectin, collagen types I and IV, laminin, and SPARC (secreted protein, acidic, and rich in cysteine). CONCLUSIONS: This study is the first description of the culture and propagation of purified retinal EC from Macaca monkey, a widely accepted model for the human retina. These cultures will be highly relevant to studies of abnormal vascular disease in the human eye.

Acetylation↗
PubMed1996

Synaptic development in macaque monkey retina and its implications for other developmental sequences.

New and existing data are presented regarding synaptic development in primate retina with the aims to identify the sequence in which individual cell types form synapses in the inner plexiform (IPL) and outer plexiform (OPL) layers; to compare synaptic development sequences in cone-dominated fovea and rod-dominated peripheral retina; to compare synaptic formation with other aspects of cell differentiation; and to explore the possible roles for synapses in development. The first synapses are formed in the foveal IPL by bipolar axons at fetal day 55, followed at fetal day 60 by cone ribbon synapses. Amacrine synapses in the foveal IPL only appear in significant numbers at fetal day 88. In peripheral retina amacrine synapses are formed at fetal day 78, bipolar at 99, and photoreceptors at 105. Thus, the fovea forms the first synapses and the IPL matures before the OPL across the retina, but the fovea has a different bipolar/amacrine sequence than peripheral retina. Foveal synapses are present before many photoreceptor-specific proteins such as opsins can be detected, suggesting that some phenotypic information from the inner retina could influence the direction of photoreceptor development. The early synaptic development in the fovea may serve an important mechanical role during subsequent cell migrations that form the mature foveal pit and tightly packed cone foveola.

Animals↗
PubMed1996

Development of astrocytes and their relation to blood vessels in fetal monkey retina.

PURPOSE: To determine the development of astrocytes and their vascular relations in Macaca monkey retina. METHODS: Sections and wholemounts of retinas from fetal day (Fd) 65 to adult animals were analyzed immunohistochemically to detect glial fibrillary acidic protein (GFAP) and vimentin. RESULTS: Astrocytes appeared first near the optic disc, then subsequently further peripherally, but avoided the fovea. In the nerve fiber layer, round and ovoid cells extended processes parallel to ganglion cell axons. In the ganglion cell layer, ovoid and stellate cells exhibited anisotropic processes or a honeycomb network. The inner lamina of astrocytes developed ahead of the outer lamina, and both reached their final positions before birth. Astrocytes lay more peripherally than did developing blood vessels, and the growing edge of nerve fiber layer vessels lay between the two astrocytic layers. Spindle cells, which may be vascular precursor cells, often aligned along linear astrocytic processes. Occasional spindle-shaped cells containing GFAP or vimentin were identified as immature glia. Astrocytes and blood vessels coincided regionally during development, but astrocyte processes were typically not in register with the meshwork of growing blood vessels. Astrocyte-vessel associations increased during fetal life and postnatally. CONCLUSIONS: During development, astrocytes display the same bilaminar pattern and morphologies present in adult retina. Astrocytes and blood vessels exhibit a similar regional distribution, but develop in distinct spatial patterns. Vessel investment by astrocytic processes increases during fetal life but is variable at all ages.

Animals↗
PubMed1995

Developmental changes in calretinin expression in GABAergic and nonGABAergic neurons in monkey striate cortex.

The development of the calcium-binding protein calretinin (CaR) and its co-localization with GABA was studied in the striate cortex of Macaca monkeys from fetal day (Fd) 45 to adult. At Fd45, early neurons resembling Cajal-Retzius cells are stained in the marginal zone (MZ). At Fd55 the MZ is filled with CaR+ Cajal-Retzius cells and their processes, and scattered CaR+ cells are also found in deep cortical plate (CP), intermediate zone (IZ), and subventricular zone (SVZ). At Fd66, a band of CaR+ fibers appears in the IZ, corresponding to the location of the geniculocortical axons. This fiber band labels heavily until Fd130 but then ceases to be immunoreactive by postnatal (P) 16 weeks. At Fd85-101, the number of CaR+ cells in the CP, SVZ, and ventricular zone (VZ) reaches its highest cell density. After Fd130, CaR+ cells are concentrated in layer II and upper layer III, and this distribution changes little into adulthood. After mid-gestation, there is a progressive loss of CaR+ cell bodies and processes in the MZ, and these are rare in the adult cortex. Just before birth, a weakly stained CaR+ cell band appears in layer IVA at the border between layer IVA and IVB, but this band disappears immediately after birth. Another CaR+ cell band appears transiently in upper layer V just below the border with layers IV at P6 months. These results suggest that CaR is expressed early in fetal development in the cell populations that are immunoreactive for CaR in the adult. However, developmental events related to cortical maturation during late prenatal and early postnatal stages result in transient expression of CaR in neurons that are not immunoreactive for CaR in the adult. CaR-immunoreactivity is colocalized with GABA in almost all CaR+ cells with the exception of Cajal-Retzius cells in the MZ and some large cells observed at Fd70-101 in the VZ. The band of CaR+ fibers in the IZ is GABA-. At Fd90, almost all (> 96%) CaR+ cells are GABA+ in the CP and the first developed layers V/VI. This percentage declines later, so that on average 80% of CaR+ cells are GABA+ in adult cortex. At Fd135, 53% of GABA+ neurons located in layers II/III are CaR+; this percentage declines to 37% in the adult. These double-label patterns suggest that early in fetal development the majority of GABA+ cells stain for CaR and that expression of CaR may be related to the migration of these neurons into the cortical plate. Once they attain their final position in the cortex many GABA+ cells loose CaR-immunoreactivity, so that in postnatal life only a minority of GABA+ neurons contain this calcium-binding protein.

Animals↗
PubMed1995

Quantitative analysis of synaptogenesis in the inner plexiform layer of macaque monkey fovea.

Synaptogenesis has been tracked by using quantitative electron microscopic methods in the inner plexiform layer (IPL) of the developing Macaca monkey fovea from fetal day (Fd) 55 to Fd132. Vesicle-containing profiles were classified according to whether (1) they contained a ribbon indicating that they originated from a bipolar cell, or (2) the profile formed a junction. Group 2 was further subdivided by morphological characteristics into (2a) amacrine, (2b) bipolar, or (2c) unknown profiles. Ribbon-containing bipolar profiles are clearly identifiable at Fd55 when they occur at a density of 0.9/100 microns2. Bipolar synapses increase rapidly to 4.7/100 microns2 by Fd88, similar to their density at Fd132. Identifiable amacrine profiles forming a junction are rare at Fd55-68. By Fd88, amacrine synaptic density has jumped to 6.7/100 microns2 and continues to increase to 9.5/100 microns2 at Fd132. These quantitative data strongly suggest that, at the Macaca fovea, bipolar synaptogenesis both begins and ends before amacrine synaptogenesis. The large number of immature amacrine synaptic profiles and densities at Fd132 suggests that amacrine synapses continue to form after Fd132. This study confirms that cone-dominated monkey fovea has a different sequence of synaptogenesis than the rod-dominated peripheral retina (Nishimura and Rakic, [1985] J. Comp. Neurol 241:420-434). The data support the concept that synaptic developmental sequence is determined by the type of photoreceptor which dominates a particular retinal region or species. Bipolar ribbon synapses are observed in the outer half of the IPL at Fd55, are present in the inner IPL at Fd60, and then, with increasing age, are found throughout the IPL. This pattern strongly suggests that vertical OFF bipolar pathways form earlier than ON pathways in the IPL. In contrast, amacrine profiles are found throughout the IPL at the youngest ages, with an adult-like banding pattern present by Fd132.

Age Distribution↗
PubMed1995

Sources of subcortical afferents to the macaque's dorsal lateral geniculate nucleus.

BACKGROUND: The dorsal lateral geniculate nucleus (dLGN) is the thalamic region responsible for transmitting retina signals to cortex. Brainstem pathways to this nucleus have been described in several species and are believed to control the retinocortical pathway depending on the state of the animal (awake, asleep, drowsy, etc.). The purpose of this study was to determine all of the subcortical sources of afferents to the dLGN in a higher primate, the macaque monkey, whose visual system is similar to that of humans. METHODS: Injections of horseradish peroxidase (HRP), with or without conjugation to wheat germ agglutinin, were made into the dLGNs of seven macaque monkeys, followed by perfusion, brain sectioning, and analyses of neurons in the brainstem, thalamus, and hypothalamus that contained the retrogradely transported marker. RESULTS: The reticular nucleus of the thalamus, pedunculopontine nucleus, parabigeminal nucleus, pretectal nucleus of the optic tract, superior colliculus, dorsal raphe nucleus, and tuberomammillary region of the hypothalamus contained many retrogradely labeled neurons ipsilateral to the injections. In the contralateral brainstem, HRP-labeled cells were found only in the pedunculopontine nucleus, nucleus of the optic tract, and dorsal raphe nucleus. The number of labeled neurons on the contralateral side was about one-half of that in corresponding ipsilateral nuclei. The locus coeruleus contained no labeled neurons in four of the macaques that had injections limited to the dLGN. CONCLUSION: There are seven subcortical regions that send afferents to the dLGNs of macaque monkeys. Except for the locus coeruleus, these are the same as observed for other species, such as the cat and rat, and indicate the possible sources of subcortical control over the dLGNs of humans.

Animals↗
PubMed1995

Transient co-localization of calretinin, parvalbumin, and calbindin-D28K in developing visual cortex of monkey.

This paper reports a double-labelling immunocytochemical study of the three calcium-binding proteins calretinin, parvalbumin, and calbindin-D28k in developing and adult Macaca primary visual cortex. In adult visual cortex, each protein marks a subset of GABAergic neurons with a characteristic laminar distribution and virtually no co-localization was found between these three proteins, suggesting that each calcium-binding protein may serve as a marker for one or more cortical subcircuits. The immature visual cortex, immunostained using identical techniques was then analysed to determine if each calcium-binding protein could serve as a developmental marker for these circuits. The Cajal-Retzius cells of layer 1 contained all three proteins during development. Calbindin-D28k and calretinin were co-localized starting at Fd (foetal day) 45 and after Fd125, parvalbumin also was present in the same Cajal-Retzius cells. All three proteins continued to be expressed until the Cajal-Retzius disappeared postnatally. In layers 2-6 calbindin-D28k and calretinin were never co-localized. In contrast, parvalbumin and calretinin were found in neurons of deep layer 3 from Fd 155 to postnatal (P6) weeks with a few persisting even later. Before birth almost all PV+ neurons in layers 4-6 were CaB+, but by P3 weeks only a few PV+/CaB+ neurons remained in layer 4C and these completely disappeared by P6 weeks. Co-localization in layer 4 neurons overlaps the period of ocular dominance segregation, suggesting that the onset of cortical maturity coincides with segregation of calcium-binding proteins within the GABA interneurons.

Animals↗
PubMed1995

The appearance of rod opsin during monkey retinal development.

PURPOSE: To determine the temporal and spatial pattern of rod opsin appearance in Macaca monkey retina. METHODS: Frozen sections from fetal day (Fd) 55 to adulthood (birth = Fd168) containing the entire horizontal meridian were stained using Rho4D2 monoclonal antiserum visualized with immunofluorescent labeling. At Fd66, Fd79, and Fd89, retinal samples taken at known eccentricities were studied from the opposite eye using standard electron microscope methods. RESULTS: Rod opsin was detected at Fd66 in or near the fovea, and a second focus appeared at Fd75 to Fd77 near the optic disc in the nasal rod ring. The earliest opsin appeared in the apical stubs, which resembled the apical connecting cilium in the electron microscope. Staining of the entire cell body membrane, including the synaptic spherule, was present 4 to 7 days later. Opsin expression had a nasal bias with rods at the nasal ora labeled at Fd140, whereas temporal ora was not labeled until Fd155. Cell body labeling disappeared by Fd132 across central retina but persisted into the first postnatal year in far peripheral retina. Outer segment (OS) length measurements showed that rods in the rod ring had the longest OS between Fd115 and postnatal week 9. Rod OS at all retinal eccentricites continued to elongate between 11 months of age and adulthood. CONCLUSIONS: Rod opsin expression follows a foveal-to-peripheral gradient beginning at Fd66 and ending near birth. Rod opsin is detected first in the connecting cilium and slightly later in the entire cell membrane, and then cell membrane labeling disappears as the heavily labeled OS elongates. Although the first OS appear on rods near the fovea, these OS still are short at birth and do not reach adult length until after 2 years of age. The longest OS at birth are found on rods at the rod ring, suggesting that this region could have higher scotopic sensitivity than central retina at birth.

Aging↗
PubMed1994

Differential localization of two glutamic acid decarboxylases (GAD65 and GAD67) in adult monkey visual cortex.

Adult monkey primary visual cortex contains a diverse population of stellate neurons that utilize the neurotransmitter gamma aminobutyric acid (GABA). Two glutamic acid decarboxylase (GAD) enzymes that synthesize GABA, GAD65 and GAD67, were localized within these stellate neurons by in situ hybridization of 35S or digoxigenin (DIG) labeled riboprobes. Double labels were done by using 35S GAD67 riboprobe and GABA immunocytochemistry on the same section to verify that the neuronal population identified by immunocytochemistry was the same one studied in the in situ hybridization experiments. We find that GAD65 mRNA and GAD67 mRNA are widely distributed in the cortex, with four bands of heavily labeled neurons in upper layer 2, lower 3, 4C, and 6. GAD67 labeled neurons were more obvious in layer 4C beta, while GAD65 containing neurons were common in layer 1 and white matter. Northern blots and in situ hybridization on sections with both 35S and DIG riboprobes indicate that cortical neurons typically contain more GAD67 mRNA. Cell counts show that 18% of all cortical neurons contain GAD67 mRNA and 13% contain GAD65 mRNA, suggesting that a small population of GABA neurons might lack GAD65. Cell bodies that contain high amounts of GAD65 mRNA are prominent in layers deep 3, 4B, 4C alpha, and 6 and often are the largest cells in their respective layers. Double labels demonstrate that 96% of all GABA+ neurons contain GAD67 mRNA. Neurons heavily labeled for GABA tend to have smaller cell bodies and contain less GAD67 mRNA, while lightly labeled GABA neurons are larger and contain more GAD67 mRNA. These data indicate that most GABA neurons in monkey striate cortex contain both GAD enzymes. Although the differences in GABA content, cell size, laminar distribution, and GAD mRNA concentration suggest different requirements for GAD67 and GAD65 in cortical circuits, our experiments do not reveal what different roles these two enzymes subserve within GABAergic stellate neurons.

Animals↗
PubMed1994

Vascular development in primate retina: comparison of laminar plexus formation in monkey and human.

PURPOSE: The temporal and spatial sequence of development of laminar vascular plexuses was determined qualitatively and quantitatively in monkey and human retina. METHODS: Histologic and cytochemical methods were used to study Macaca monkey eyes from fetal day 55 (F55d; birth = F168d) to 17 years, and human retina from fetal 21 weeks to adult. RESULTS: In monkey retina, spindle-shaped, presumed vascular precursor, cells appear at F55d in the nerve fiber layer (NFL) adjacent to the optic nerve. The vascular plexuses in the NFL-ganglion cell layer appear first and form in the presence of spindle cells. Nerve fiber layer vessels extended radially to reach the temporal ora at F95d and nasal ora at F110d. The capillary plexus at the inner border of the inner nuclear layer (INL) appears at F120d near the optic disc, whereas the plexus at the outer INL border appears at F130d. Both reach their final position before birth. The INL plexuses form by endothelial budding from more vitread vessels in the absence of spindle cells. In the NFL, vessel growth to match retinal growth at the ora also involves endothelial budding. The growth rate of all plexuses was approximately 225 microns/day. The central fovea and the most peripheral retina adjacent to the ora serrata remained avascular throughout development. Differences between humans and monkeys include: Human vessels complete maturation after birth; human vessels reach the nasal ora earlier than the temporal ora; and spindle cells are more abundant and dispersed over a greater area within human NFL. Growth rates of human plexuses were comparable to those in monkeys. CONCLUSION: In both primates, deeper capillary plexuses form only by extension from existing vessels (angiogenesis). In the NFL, early vessel formation involves spindle precursor cells (vasculogenesis). The main difference between monkey and human in these processes is that the mature monkey vascular pattern is established well before birth.

Adolescent↗
PubMed1993

A comparison of the development of neuropeptide and MAP2 immunocytochemical labeling in the macaque visual cortex during pre- and postnatal development.

The appearance of Substance P (SP) and Neuropeptide Y (NPY) has been studied using light microscopic immunocytochemical labeling throughout the complete developmental span of Macaca nemestrina monkey striate cortex. In the adult, 80% of the NPY+ neurons occur in the white matter (WM) and most of the remainder are medium to large multipolar neurons in layer 2. Fibers occur in all layers except 4C and are very numerous, given the relatively small number of NPY+ cell bodies. NPY+ neurons first were seen at embryonic day (E) 75. Most neurons were in the intermediate zone (IZ), but a few were in the immature cortical plate (CP). An adult-like distribution was present by E125 for neurons and by birth for fibers, but fiber staining intensity and number increased to postnatal year 1 (P1yr). In adult cortex, numerous SP+ nonpyramidal neurons were present in layers 2-6 and WM, but SP+ fibers were surprisingly infrequent. During development, significant numbers of SP+ neurons were not seen in the CP until E113-125. Later prenatal ages had a prominent plexus of SP+ cell bodies and fibers at the layer 5/6 border. This plexus disappeared by P12wk due to either down-regulation of SP or cell death. SP+ neurons in IZ/WM were very sparse until birth after which they increased in number and staining intensity up to P1yr, suggesting a postnatal up-regulation of SP in a preexisting WM subpopulation. Cell densities were determined for SP, NPY, and the neuron-specific marker microtubule-associated protein 2 (MAP2) to clarify the developmental dynamics of IZ/WM neurons. MAP2+ cell densities in WM peaked around birth and then declined 20% in the outer half and 77% in the inner half of WM. SP+ cell density rose 57% from birth to P20wk and then declined 20% into adulthood. NPY+ cell density was fairly constant prenatally and then rose 300% by adulthood. Neuropeptide cell density changes took place predominantly in the outer WM. These data indicate that cell death does occur in the general population of monkey striate cortical WM neurons. In contrast, both SP+ and NPY+ cells are characterized by minimal cell death and a late expression of neuropeptides which causes an increase in neuropeptide+ cell density in postnatal WM.

Animals↗
PubMed1993

Comparison of immunolocalization patterns for the synaptic vesicle proteins p65 and synapsin I in macaque monkey retina.

The distributions of the two synaptic vesicle proteins p65 [Matthew et al. (1981) J. Cell Biol., 91:257-269] and synapsin I [De Camilli et al. (1983) J. Cell Biol., 96:1337-1354] were compared in macaque monkey retina using pre-embedding immunocytochemistry for both light and electron microscopy. The monoclonal antibody AB-48 against p65 labeled ribbon-containing synaptic terminals of cone, rod, and bipolar cells as well as many conventional synapses of amacrine cells. In contrast, a polyclonal antiserum against synapsin I (SYN I) labeled many amacrine conventional synapses but no photoreceptor or bipolar ribbon synaptic terminals. Horizontal cell pre- and post-synaptic profiles in the outer plexiform layer were not labeled by either antibody. At the light microscopic level, the banding patterns in the inner plexiform layer also differed for the two antibodies, with four bands of AB-48 immunoreactivity in sublayers S1, S2, S4, and S5 but only three bands of SYN I immunoreactivity in S1, S3, and S5. SYN I also labeled varicose fibers in both the inner nuclear layer and the outer plexiform layer that are probably processes of dopaminergic and GABAergic interplexiform cells. Varicose fibers in the ganglion cell layer were labeled by both antibodies. These results provide the first electron microscopic immunocytochemical labeling for AB-48 and SYN I in intact retina and confirm that AB-48 labels both ribbon and conventional synaptic terminals, whereas SYN I labels only conventional synapses.

Animals↗
PubMed1993

Immunocytochemical localization of GABA and glycine in amacrine and displaced amacrine cells of macaque monkey retina.

Recent studies have varied widely in the percentages of GABA- and glycine-immunoreactive (GABA+, GLY+) amacrines reported for primate retina. We compared the distributions of GABA+ and GLY+ amacrines and displaced amacrines at seven locations along the horizontal meridian of macaque retina using postembedding immunogold labeling with silver intensification. The percentage of GABA+ amacrine profiles was higher in central retina (50-55%) than peripheral retina (30-40%), whereas the percentage of GLY+ amacrine profiles did not vary much with eccentricity (52-57%). GABA and glycine were colocalized in 5-20% of amacrines, depending on the eccentricity, whereas 5-30% of amacrines were not immunoreactive for either neurotransmitter. GABA+ amacrines were slightly larger than GLY+ amacrines or Müller cells. In the ganglion cell layer, 5-20% of neurons were labeled for either GABA or glycine and were identified as displaced amacrines. Of these, 53% were GABA+ only, 11% were GLY+ only, and 37% were double-labeled. A few large, very lightly labeled GABA+ cells were identified as ganglion cells. Other features that varied with eccentricity included the linear density of GABA+ and GLY+ amacrines, and the ratio of amacrines to Müller cells.

Animals↗
PubMed1992

VVA-labelled cells in monkey visual cortex are double-labelled by a polyclonal antibody to a cell surface epitope.

The staining patterns produced by the lectin Vicia villosa and by a commercially available polyclonal antibody generated to substance P were analysed and compared in monkey visual cortex at the light and electron microscopic levels. Vicia villosa lectin labels the cell surface of a subpopulation of cortical cells, producing a meshlike pattern over the soma and proximal dendrites. The polyclonal antibody labels three distinct elements in the cortex: a pericellular epitope present on a subpopulation of non-pyramidal cells, and putative intracellular sites in a type of small pyramidal cell located at the layer 5/6 border, and in a small number of non-pyramidal cells in the underlying white matter. Because of the similarity of the appearance of the Vicia villosa lectin labelling and the pericellular labelling produced by the polyclonal antibody, further experiments were conducted to determine the relationship between the cell surface sites recognized by these markers. Double-labelling experiments show that both sites are present on the same population of cells, and at the ultrastructural level both markers appear to outline the intersynaptic cell membrane, sometimes extending around presynaptic elements. However, preadsorption experiments indicate that the markers recognize different sites on the cell membrane. Preadsorption experiments also show that the pericellular epitope recognized by the polyclonal antibody is unlikely to be substance P, but it may be structurally similar to keyhole limpet haemocyanin. Comparison of cortical and subcortical staining patterns produced with the polyclonal antibody and with a commonly used monoclonal antibody to substance P reveal that one of the putative intracellular epitopes recognized by the polyclonal antibody is likely to be substance P.

Animals↗
PubMed1992

Development of synapses in macaque monkey striate cortex.

A quantitative electron-microscopic (EM) analysis of the development of synaptic density (number of synapses/100 microns2 neuropil) has been done in primary visual cortex (striate, area 17) of the Old World monkey Macaca nemestrina. A comparative EM morphological study of developing synaptic contacts also was done in the same tissue. We find that a few immature synaptic contacts are present at fetal (F) 75 days either in the marginal zone, which becomes layer 1, or in the deepest portion of the cortical plate, the future layer 6. At F90-140 days synaptic contacts are found throughout the cortical plate, but their density remains higher in lower cortical layers. By F140 days synaptic density averaged for all layers (10.9) is three times higher than at F90 days. Just before and after birth, synaptic density rises very rapidly to peak at postnatal (P) 12 weeks (63) and then declines slowly to reach adult values (37.7) between 2-6 years. This pattern was further tested by comparing synaptic density in layer 2 which contains the last cells generated in the striate cortex to that in layer 6 which contains the first cells generated in the striate cortex. Layer 6 contained the first synapses, and had a higher density up to F140 days (an "inside-to-outside" distribution). Synaptic density was equal in the two layers at F152 days and P2 days, but by P12 weeks synaptic density in layer 2 was 27% higher than that in layer 6 (an "outside-to-inside" distribution). After P12 weeks, the synaptic density declined 51% in layer 2 and 21% in layer 6 so that both layers achieved similar densities by P6 years. A light and EM comparison of neuropil and synaptic contact morphology finds that, at each age up to birth, synapses in layer 2 are generally less mature than those in layer 6, but these differences disappear shortly after birth. Between P6-24 weeks, synaptic contacts throughout the cortex acquire a mature morphology that clearly differentiates between asymmetric and symmetric types, although asymmetric contacts continue to acquire more postsynaptic density until adulthood. This complex developmental pattern suggests a sequence for synaptic developments which is more related to neuron birthdate than to the arrival of extrinsic pathways or developmental events occurring in specific laminae.

Animals↗
PubMed1991

Development of the calcium-binding protein parvalbumin and calbindin in monkey striate cortex.

The development of immunoreactivity for the calcium-binding proteins parvalbumin (PV) and calbindin-D28K (Cal) was studied in Macaca nemestrina striate cortex from fetal (F) 60 days to postnatal (P) 5 + years. We correlated changes in PV and Cal staining patterns with the well-documented developmental sequence for primate striate cortex neuron generation and maturation, synaptogenesis, and thalamocortical axon interactions in an attempt to deduce a functional role for these proteins. Our major findings is that Cal and PV have diametrically opposed developmental patterns except in layer 1. At F60 days both are present only in neurons of layer 1 and the number of labeled cell bodies and processes increases up to F125 days. Almost all Cal+ and PV+ cells in layer 1 disappear by P12 weeks. Cal is present by F113 days in pyramidal and stellate neurons, particularly layers 4-6. The numbers and staining density of cells in layers 2-6 increases up to birth and then both decline by P9-12 weeks. Supragranular layers show a second increase in Cal labeling from P20-36 weeks, and then there is a slow decline to the adult pattern which is reached by P1-2 years. Cell bodies in layers 4A, 4C alpha, and deep 4C beta are heavily Cal+ during pre- and early post-natal periods, but upper 4C beta remains unlabeled. PV is not seen until F155-162 days in layers 2-6. Large stellate and a few pyramidal cells appear first in layers 5/6 and 4C alpha, but PV+ stellate neurons are found in all layers except 4C beta by P6 weeks. Layer 4C beta contains a few PV+ cell bodies at P3 weeks, and light neuropile staining at P6 weeks, but then PV labeling rapidly increases so that by P12 weeks the density of 4C beta exceeds that of 4C alpha. Striate cortex has an adult pattern of cell number and neuropile density by P20 weeks. These developmental patterns suggest that the highest density of Cal cell body staining does not correlate with synaptogenesis, or the postnatal critical period of visually driven, binocular interactions. Rather Cal appears when lateral geniculate axons arrive in cortex, persists over the entire span of thalamocortical interactions, and disappears during the decline of cortical plasticity. The appearance of PV is highly correlated with the onset of complex visually driven activity at birth, while both the number of PV+ cell bodies and the density of PV+ neuropile reach adult levels coincident with the completion of thalamocortical connections.

Animals↗