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Ovariectomized rabbits received 3H-estradiol via an ear vein and were killed one hour later. Autoradiograms were prepared and exposed up to six months. Labeled cells, as indicated by many silver grains over the nucleus of a neuron, were found in many nuclei of the brain. Thus, the bed nucleus of the stria terminalis had labeled cell bodies. The stria terminalis leads into the medial preoptic area where great numbers of cells concentrated the estrogen. Farther into the hypothalamus the labeled cells were numerous in the ventromedial and arcurate nuclei. Other locations with labeled cells were the lateral septal nucleus and nucleus accumbens septi, the periventricular preoptic nucleus, anterior hypothalamic nucleus, nucleus supraopticus diffusus, posterior hypothalamic nucleus, and premammillary nucleus. The labeled cells could be followed into the central gray surrounding the aqueduct of Sylvius. The amygdaloid nuclei, and in particular the medial amygdaloid nucleus, had labeled cells as did the most ventral posterior part of the hippocampal cortex. The results are discussed in comparison with those in the rat, and with reference to physiologic data.
Anat Rec 1975 Feb
PMID:Localization of cells retaining 3H-estradiol in the forebrain of rabbits. 111 57

Mouse anterior pituitary cells cultured for 2 days were stimulated with one of three biotinylated-GnRH probes ([biotinyl-Lys6]-[D-Lys6]GnRH, [biotinyl-Ser4]-[D-Lys6]GnRH, [biotinyl-Ser4]-[D-Trp6, des-Gly10]GnRH) in the cold (4 degrees C) for 1 hr. These cells were subsequently fixed and an avidin-gold complex was conjugated to the bound GnRH. After a second fixation, the gold label was silver-enhanced for viewing with a scanning electron microscope. Gonadotropes were identified as a result of the labeling procedure, measured for size, and the number of GnRH receptor sites counted. Gonadotropes ranged from 3 to 13 microns in diameter and contained from 23.2 +/- 3.3 to 338.4 +/- 25.2 sites per cell depending upon the size of the cell and the ligand employed. The methods described should be applicable for studying the topographical distribution of a variety of cellular receptors.
Anat Rec 1992 May
PMID:Topographical mapping of GnRH receptors on dispersed mouse pituitary cells by backscattered electron imaging. 131 44

The movement of proteins into and out of enamel was followed over time using a highly sensitive microprecipitation technique to quantify the amount of TCA-insoluble radioactivity present within small pieces of freeze-dried enamel and cells (enamel organ) dissected from the mandibular incisors of rats injected with L-[35S]-methionine. Conventional image processing techniques were also used to estimate the number of silver grains over enamel and cells in radioautographs of mandibular incisors from rats similarly injected with L-[methyl-3H]-methionine. Data from both techniques indicated that the average half-life for labeled proteins secreted into enamel was about 8.9 days. Typically, radioactive proteins accumulated in increasing amounts for 8 hours after which they were lost slowly up to 4 days and more rapidly thereafter when enamel formed during the secretory stage underwent maturation. The half-life for radioactive proteins in cells was only about 20.7 hours. No significant accumulation of radioactivity could be detected in the TCA-soluble or TCA-insoluble fractions of cells as enamel development proceeded. Results from this study suggest that radioautographs provide an accurate estimate of changes occurring to proteins in enamel and cells except at early time intervals (less than 1 hour) when a high percentage of total radioactivity is present within the TCA-soluble fraction of cells.
Anat Rec 1992 Jan
PMID:Correlated biochemical and radioautographic studies of protein turnover in developing rat incisor enamel following pulse-chase labeling with L-[35S]- and L-[methyl-3H]-methionine. 153 54

Sensory nerve fibers of the cochleo-vestibular ganglion (CVG) innervate the otic epithelium in the early chick embryo by directed growth. To see if the target tissue could exert a tropic influence, we co-cultured CVGs from chick embryos (Hamburger-Hamilton stages 16-30) in a 3D collagen matrix with their normal target epithelium or with other epithelial tissues taken from the same or different stages of development. The pattern of neurite outgrowth and the viability of the CVG after five days in vitro were assessed histologically with a silver method. On the basis of the patterns of neurite outgrowth directed toward the epithelium, the cultures were classified as having slightly, mostly, exclusively, or no directed outgrowth. Of 49 cultures containing otic epithelium, 33 had mostly or exclusively directed growth patterns. This effect did not depend on any particular stage difference between co-cultures or on their viability in vitro. Cultures of non-sensory otic epithelium (endolymphatic duct) also presented directed growth patterns. Co-cultures with ectoderm from forelimb or visceral arch had little, if any, directed growth. The directed growth could not be explained simply as a result of guidance by non-neuronal cells or of the viability of the explants. The results are consistent with the hypothesis that the otic epithelium provides a tropic factor that attracts growing CVG fibers.
Anat Rec 1992 Feb
PMID:Tropic effects of otic epithelium on cochleo-vestibular ganglion fiber growth in vitro. 154 5

Forty-nine 12-week-old male Syrian golden hamsters (Mesocricetus auratus), weighting 108 to 128 g, were injected i.p. with [3H]-thymidine (3HTdR 2 microCi/g body wt). Animals were divided into 7 weight-matched groups and were killed at 1 hour (day zero) and 1, 2, 3, 4, 7, and 14 days after thymidine injection. Lungs were fixed by vascular perfusion of 4% formalin/1% glutaraldehyde in 0.2 M cacodylate buffer at pH 7.4, cut at 2 microns, dipped in Kodak NTB3 or NTB2 nuclear emulsion, exposed for 2 weeks, developed and stained. In each airway cross-section, total epithelial and fibroblast counts and labeled cell count were estimated. A cell was considered labeled when 3 or more silver grains appeared on its nucleus. The background grain count was less than 1 grain per nucleus. Mean epithelial and fibroblast cell density in a 100 microns segment were respectively 14.8 +/- 0.1 and 6.6 +/- 0.1 cells (the second number is one standard error of the mean). One hour after labeling, their respective labeling indices (L.I.) were 0.13 +/- 0.02 and 1.24 +/- 0.1. On day 1, their L.I. doubled and then returned to the initial value. One hour after labeling, epithelial and fibroblast mean grain counts did not differ significantly. They were respectively 20.6 + 1, and 15.8 +/- 2. Because grain count intensity is closely related to DNA synthetic time, it seems plausible that epithelial and fibroblast synthetic times do not differ much. Cell turnover and cell cycle times were estimated from grain count dilution curves.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1992 Jun
PMID:Cell kinetics of lamina propria fibroblasts in normal adult hamster bronchus. 160 89

The cytoplasmic distribution of poly(A)+ mRNA and its relationship to annulate lamellae were examined in developing Necturus maculosus oocytes by in situ hybridization with [3H]poly(U). The specificity of [3H]poly(U) binding was tested by incubating control ovarian sections with either KOH or RNase A before in situ hybridization. In both experiments, the silver grain densities were markedly reduced. Poly(A)+ RNA is uniformly distributed in the cytoplasm until the mid-growth phase and then later in vitellogenesis becomes localized in the subcortical ooplasm. The silver grain density in the cytoplasm varied during oogenesis and was greatest in previtellogenic oocytes. Annulate lamellae commonly are observed with the light microscope in oocytes prior to vitellogenesis. In such oocytes, the labeled mRNA probe is observed over cytoplasmic regions of annulate lamellae. The results suggests that a differential localization of messenger RNA occurs during oogenesis in Necturus maculosus. Furthermore, poly(A)+ RNA is present in cytoplasmic regions of annulate lamellae.
Anat Rec 1991 Jun
PMID:Cytoplasmic distribution of poly(A)-containing RNA in developing Necturus maculosus oocytes with reference to annulate lamellae. 171 57

The anatomical relationships between sensory afferents within a topographic map in the cricket cercal sensory system were studied using a computer-based reconstruction system developed in our laboratory. Individual identified mechanosensory afferents were characterized physiologically, stained with cobalt, silver intensified, and reconstructed in three dimensions. All reconstructions were scaled to a common standard. The results indicate that there is very little variability in the position or extent of the terminal arborization of identified mechanosensory afferents. The topographic map was divided relatively equally into four regions representing each of the four classes of afferents studied. These regions were discrete but not completely segregated. Approximately 30% of the topographic map contained regions of overlap between two or more classes or afferents.
Anat Rec 1991 Dec
PMID:Anatomical relationships between sensory afferent arborizations in the cricket cercal system. 172 59

Dopamine D2 receptor mRNAs have recently been cloned and their gross distribution in the central nervous system described. Quantitative in situ hybridization histochemistry with a cRNA probe complementary to the mRNAs encoding approximately 70% of the third intracellular loop of the rat D2 receptor was performed on sections of rat brain to determine whether differences previously observed in the density of ligand binding sites in subregions of the striatum were related to differences in mRNA levels. Film autoradiographic analysis demonstrated 30% more hybridization signal in the lateral compared to the medial caudate-putamen, a distribution parallel to that of binding of ligands specific for the D2 receptor. Inspection at the cellular level using emulsion autoradiography also indicated a differential distribution of the D2 receptor mRNA. Fewer positively labelled cells, as well as fewer silver grains per cell, were seen in the medial compared to the lateral half of the striatum. This suggests that the gradient seen in autoradiographic studies of the distribution of D2 receptors is related both to regional differences in D2 mRNA levels and to the density of cells expressing the receptor. In addition, the distribution of cells expressing D2 receptor mRNA in the extrastriosomal matrix was compared to that in striosomes identified by the presence of a high density of 3H-naloxone binding sites. Labelled cells were mainly found in the matrix (3H-naloxone binding-poor) but were also seen in striosomes (3H-naloxone binding-rich). The results suggest that differences in levels of D2 binding sites in subregions of the striatum are related to differences in the level of expression of this receptor in intrinsic striatal neurons, suggesting differential regulation of dopamine D2 receptor gene expression in topographically distinct striatal neurons.
Anat Rec 1991 Dec
PMID:Heterogeneous distribution of dopamine D2 receptor mRNA in the rat striatum: a quantitative analysis with in situ hybridization histochemistry. 183 6

We used indirect immunofluorescence and immunogold light microscopy to examine the distribution of hepatic lipase, an enzyme involved in lipoprotein metabolism, in ovaries of gonadotropin-treated immature rats. Antibodies utilized were rabbit anti-rat hepatic lipase IgG, anti-rat von Willebrand factor (VWF, an endothelial cell marker), and goat anti-rabbit IgG conjugated to gold particles or rhodamine. Immunoreagents were applied to fresh frozen sections of unfixed ovary or liver (positive control) or were delivered to ovaries by vascular perfusion before fixation in situ and silver-enhancement of sections. Appropriate controls verified that the immunolocalizations were specific. Immunofluorescence implied that luteal but not stromal blood vessels of ovaries were positive for hepatic lipase, whereas luteal and stromal blood vessels bore VWF. The improved morphology gained by perfusing ovaries with antibodies allowed precise localization of the enzyme. Hepatic lipase was concentrated within thin-walled vessels of corpora lutea but not those of stroma in ovaries at the time of peak steroidogenic activity. Quantification of hepatic lipase-labeled vessels in stromal and luteal compartments confirmed our visual impression. Many images suggested that stromal vessels lacking hepatic lipase gained this enzyme upon contact with luteal tissue. Perfusion of ovaries with cationized ferritin labeled all ovarian vessels equally well, ruling out the possibility that the observed distribution of hepatic lipase was artifactual. These findings demonstrate that ovarian blood vessels are heterogeneous for hepatic lipase. Moreover, they imply that luteal tissue, perhaps luteal cells, may influence expression of hepatic lipase binding sites by endothelial cells.
Anat Rec 1991 Jul
PMID:Heterogeneity among ovarian blood vessels: endogenous hepatic lipase is concentrated in blood vessels of rat corpora lutea. 186 6

The conduction velocity and histological structure of motoneurons innervating normal and hypertrophied rat plantaris muscles were investigated. Hypertrophy was produced by ablation of synergist muscles. Single motor units were obtained by ventral root dissection and conduction velocities measured. The structure of neurons was investigated following retrograde labeling with horseradish peroxidase. A combined silver, gold and cholinesterase staining method was developed to study the motor endplate. In addition, the peripheral nerve was fixed, embedded in Araldite, and sectioned for determination of axonal size and myelin thickness. Conduction velocity of motor axons decreased following hypertrophy of the skeletal muscle (control CV = 75.8 +/- 8.9 m s-1, n = 94, hypertrophy CV = 69.0 +/- 12.3 m s-1, n = 84). However, no alteration in the size of motor axons or myelin thickness could account for this alteration in conduction velocity. Mean motoneuronal soma size decreased following muscle hypertrophy (soma diameter: control 36.1 +/- 4.6 microns, n = 283, hypertrophy 32.9 +/- 4.5 microns, n = 294). The complexity of the motor endplate increased following hypertrophy with an increased occurrence of nodal sprouts. In addition, the area of cholinesterase staining increased following hypertrophy (control 588.1 +/- 297.2 microns 2, n = 269, hypertrophy 857.7 +/- 357.0 microns 2, n = 269). This study found that both the morphological and physiological parameters of motoneurons innervating a hypertrophied muscle were shifted toward those of normal rat slow motor units.
Anat Rec 1991 Jan
PMID:Functional and structural changes of rat plantaris motoneurons following compensatory hypertrophy of the muscle. 199 79


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