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The present peroxidase-antiperoxidase immunohistochemical study demonstrated that approximately 50% of the total chromaffin cells of the rat adrenal medulla exhibited NPY-like immunoreactivity. The immunoreactive material was localized in the core of the chromaffin granules as well as diffusely in the cytoplasm. By combination of immunohistochemistry with noradrenaline-fluorescence microscopy, all NPY-immunoreactive chromaffin cells are nonfluorescent, indicating that all NPY-chromaffin cells co-store adrenaline. A comparison of two consecutive sections, each of which was processed for the immunostaining with anti-NPY and anti-Met-Enk-Arg-Gly-Leu antisera, respectively, indicated that NPY and preproenkephalin A and its derivatives coexist in approximately one-fifth of the total NPY-immunoreactive cells. In addition to the NPY-immunoreactive cells, a plexus of NPY-immunoreactive nerve fibers with varicosities was found in the subcapsular regions of the adrenal gland. The nerve fibers were often associated with small blood vessels and extended into the zona glomerulosa. Single NPY-immunoreactive fibers were sparsely distributed in the deeper regions of the cortex and in the medulla. Ganglion cells in the adrenal gland were not seen exhibiting intensely positive NPY-like immunoreactivity. The NPY-immunoreactive nerve fibers contained abundant small clear vesicles mixed with a few small and large granular vesicles. The immunoreactive material appeared on the granular cores as well as in the axoplasm. The NPY fibers were closely apposed to smooth muscle cells and pericytes of small blood vessels in the cortex.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1986 Mar
PMID:Neuropeptide tyrosine (NPY)-like immunoreactivity in adrenal chromaffin cells and intraadrenal nerve fibers of rats. 351 14

The antigenic phenotype of mouse lymph node follicular dendritic cells (FDCs) was studied by immunocytochemical techniques. Indirect fluorescence was used in conjunction with monoclonal antibodies to localize FDC surface antigens on FDC-enriched cell preparations and in cryostat sections. Lymph nodes from rats and mice were also labeled directly for Ia antigens with fluorescein- or peroxidase-conjugated Ia-specific monoclonal antibodies (i.e., MRC Ox4 and 10-2.16, respectively). Lymphoid tissue was also prepared for electron microscopy to allow clear distinction between Ia antigens of B lymphocytes and FDCs in situ. In these experiments, gold-labeled antigen was used to clearly identify FDCs and their processes among the Ia-positive cells of lymph node follicles. The labeling observed by light and electron microscopy showed that FDCs expressed Ia in situ and in vitro. Additional surface determinants shown to be expressed by FDCs included H2-K, common leukocyte antigen, and the receptor for the Fc portion of IgG1 and IgG2b. Neither macrophage antigens, such as Mac-1, Mac-2, Mac-3, and F4/80, nor the lymphocyte markers Ly-1, Ly-2, and Thy-1 were expressed by FDCs. Thus, the antigenic phenotype of FDCs, along with their distinctive dendritic morphology, their nonphagocytic and nonadherent nature, and their ability to trap and retain immune complexes on their plasma membrane, identifies them as a unique cell population.
Anat Rec 1986 Jul
PMID:Antigenic phenotyping of isolated and in situ rodent follicular dendritic cells (FDC) with emphasis on the ultrastructural demonstration of Ia antigens. 352 78

Human NK activity is known to be associated with a population of large granular lymphocytes (LGL) exhibiting several immunophenotypic surface markers including Leu-11a (NKP-15), Leu-7 (HNK-1), Leu-3a (T4), and Leu-2a (T8). Based upon correlation with cytolytic activity, Leu-11a is now considered the most specific antigenic marker for human NK cells. Present investigation compared the ultrastructure of cells expressing Leu-11a, Leu-7, Leu-3a, and Leu-2a, both in human peripheral blood lymphocytes (PBL) and the purified LGL fraction. Subcellular cytochemical reactions were investigated in Leu-7+ or Leu-11a+ PBL or LGL and in cells conjugated with K562 targets (indicating NK cytolytic potential). The surface markers, localized with monoclonal antibodies, were detected by immunoelectron microscopy by using direct or indirect avidin-biotin-peroxidase (ABC) or colloidal gold methods. A peroxidase-colloidal gold double-labeling system was used to identify subsets of Leu-7+ or Leu-11a+ cells. Previously described ultrastructural features of LGL including a villous surface, reniform nuclei, low nuclear/cytoplasm ratios, and abundant cytoplasm with vesicles, vacuoles, electron-dense granules, parallel tubular arrays (PTA), or paracrystalline inclusions were associated with Leu-7+, Leu-11a+, Leu-7+/Leu-11a+, Leu-7+/Leu-11a-, and Leu-7-/Leu-11a+ PBL or LGL. Results showed that the Leu-7+/Leu-11a+ cells were the most abundant NK cells in PBL. Lymphocyte subsets with Leu-3a or Leu-2a surface marker showed some ultrastructural features including PTA similar to Leu-7+ cells and Leu-11a+ cells, and their subsets. These T-cells appeared ultrastructurally more similar to the Leu-7+/Leu-11a- subset. Cytochemical studies showed that electron-dense cytoplasmic granules and PTA typical of the Leu-11a+ cells and Leu-7+ cells contained glycoprotein, acid phosphatase, and arylsulfatase. Large cytoplasmic vacuoles were heterogeneous and typically contained electron-dense material with DAB reactivity, membranous material, PTA, and/or paracrystalline inclusions. Glycoprotein, acid phosphatase, and arylsulfatase, and peroxidase reactive material were also found in these vacuoles. These features suggested that the vacuoles could be secondary lysosomes. The coexistence of intact PTA or degenerating PTA in the same vacuoles with paracrystalline inclusions suggested that the latter are possibly derived from PTA.
Anat Rec 1987 Mar
PMID:Immunoultrastructural studies of human NK cells: I. Ultracytochemistry and comparison with T cell subsets. 355 61

The binding of NK cells to a target cell appears to be a necessary step for NK cell-mediated cytolysis. In this report, we demonstrated effector-target binding by immunoelectron microscopy by using monoclonal antibodies against NK cells (Leu-7, Leu-11a) and T-cell subsets (Leu-2a/T8, Leu-3a/T4). The surfaces of NK and K562 cells were characterized by antitransferrin receptor antibody and various lectins. In addition, the controversial phagocytic activity of NK cells was studied by incubation of peripheral blood mononuclear cells with opsonized Staphylococcus aureus and labeling with anti-Leu-7 or anti-Leu-11a antibody. Results showed that only Leu-11a+ cells displayed a broad cell-to-cell contact with the target by a shallow intercellular interdigitation of cytoplasmic projections, while Leu-7+, Leu-2a+, or Leu3a+ cells showed only a partial contact with target without interdigitation. The Leu-11a+ cells were frequently observed in small clusters and in close association with monocytes. Cluster formation and association with monocytes were not observed in other NK and T-cell immunophenotypes. In Leu-11a+ cells conjugated with target cells, membrane-bound granules, small vesicles, parallel tubular arrays, Golgi apparatus, endoplasmic reticulum, and small vacuoles were evident and concentrated toward the target. The surface of NK cells was intensely stained for glycoprotein by chromic acid-phosphotungstic acid, whereas target cells were not stained. Transferrin receptors were stained only on the surface of target cells. Only the lectins RCA and UEA labeled the surfaces of both NK and target cells. Phagocytic vacuoles containing cell debris or fragments and ingested bacteria were found in the cytoplasm of Leu-11a+ cells but not in Leu-7+ cells. NK cells were also found within the cytoplasm of K562 target cells. All these findings suggest that Leu-11a+ cells are the true functional NK cells involved in NK cell-mediated cytolysis, phagocytosis, and emperipolesis. Therefore, the NK cell is probably "a phagocyte in lymphocyte's clothing." The presence of peroxidase in the small vesicles of NK cells and endocytotic vesicles of target cells at the effector-target contact area indicates that cytolytic enzymes or factors derived from NK cells may be transported into the target by endocytosis.
Anat Rec 1987 Mar
PMID:Immunoultrastructural studies of human NK cells: II. Effector-target cell binding and phagocytosis. 357 43

Since the number of motor neurons supplying a muscle graft is reduced, the peripheral field for the surviving motor neurons would be enlarged. This possible change in motor unit size may result in morphologic changes in the size of motor neurons within the motor neuron pool of the graft. The extensor digitorum longus (EDL) muscle from 19 transplanted and 17 age-matched normal 129 ReJ female mice were injected with horseradish peroxidase in order to examine the cell size distributions of the motor neuron pools supplying these muscles. Computer-assisted morphometric analysis of cell sizes within the motor neuron pool to the transplants indicated a significant shift in cell size, with the largest areas ranging between 630 and 1250 micron2. A number of the alpha neurons supplying the grafts were twice the average cell size for the control population (means = 592 micron2). The increase in the number of large motor neurons indicates an hypertrophy of neurons reinnervating the grafts. The long-term graft is reinnervated by a decreased population of motor neurons (means = 7), which vary in size from small to very large, reflecting both changes in the possible source of the nerve reinnervating the graft as well as alteration in the size of the motor unit, respectively.
Anat Rec 1987 Apr
PMID:Morphometric analysis of neuronal soma size within the motor nucleus of a transplanted murine skeletal muscle. 359 67

A capture enzyme immunoassay was developed for the detection of Mycoplasma bovis antigens in bull semen or preputial washings. IgG prepared from rabbits immunised with M bovis was passively adsorped to 96-well polystyrene plates. This antibody captured M bovis antigens which were then detected by using an IgG preparation from an immunised cow and murine monoclonal antibody to the bovine L-chain conjugated with horseradish peroxidase. The sensitivity of the assay was approximately 200 colour changing units (ccu)/ml and the specificity was excellent in that other species of mycoplasma, ureaplasma or acholeplasma did not react. A blind study of bull semen experimentally contaminated with M bovis detected all specimens with more than 200 ccu/ml.
Vet Rec 1987 Jun 20
PMID:Enzyme immunoassay for detection of Mycoplasma bovis antigens in bull semen and preputial washings. 362 62

Tuft cells are present in most columnar epithelia derived from endoderm including the small intestine. They are characterized by long, wide apical microvilli and an extensively developed cytoplasmic tubulovesicular system. We examined in detail the structural features of the apical plasma membrane of small intestinal tuft cells from adult guinea pigs, rats, and adult and suckling mice with freeze-fracture and conventional transmission electron microscopy methods and utilized cationized ferritin and horseradish peroxidase as tracers to determine whether tuft cells endocytose macromolecules. The microvillus membrane of intestinal tuft cells has few P-face intramembrane particles, displays little alkaline phosphatase activity, and is highly enriched in cholesterol. Tuft cell tight junctions resemble those of absorptive cells in strand count and strand-to-strand crosslinks but, unlike those of absorptive cells, they display many abluminal free-ending strands. Tuft cells of adult and suckling mouse intestine show no evidence of internalization of cationized ferritin or, in suckling mice, uptake of horseradish peroxidase. We conclude that the microvillus membrane of small intestinal tuft cells is protein-poor but cholesterol-rich and that small intestinal tuft cells do not endocytose macromolecules in bulk from the intestinal lumen.
Anat Rec 1987 Sep
PMID:Structural features of the apical and tubulovesicular membranes of rodent small intestinal tuft cells. 368 63

Specific antisera raised against calbindin-D28K (CaBP), the vitamin D-dependent calcium-binding protein from chick intestine, was used to localize the protein in chick ultimobranchial glands (UB glands) by the peroxidase-antiperoxidase technique. CaBP was localized in secretory cells in the cell cords and in a few cells of the epithelium lining the follicles. It was not found in the fibroblastlike cells in the cell cords nor in islands of parathyroid tissue present in the UB gland. The immunomarker for CaBP was distributed throughout the cytoplasm and nucleus of the secretory cells. The same cells demonstrated a positive reaction in their cytoplasm when reacted with an antiserum specific for salmon calcitonin (CT), thus confirming the presence of CaBP and CT in the same UB-gland secretory cells. In other tissues, the presence of CaBP is regarded as an end-organ marker for actions of the vitamin D endocrine system. This novel demonstration of CaBP in UB-gland cells responsible for secretion of calcitonin suggests a direct effect of the vitamin D endocrine system on those cells in addition to an indirect effect through the stimulation produced by elevated circulating calcium levels.
Anat Rec 1987 Sep
PMID:Localization of calbindin-D28K in calcitonin containing cells of chick ultimobranchial glands. 368 64

An indirect immunohistochemical method in which an avidin-biotinylated horseradish peroxidase complex is bound to the secondary antibody was used to visualize vasoactive intestinal peptide-immunoreactive (VIPI) nerves in the rat kidney. Rats were perfused with 4% paraformaldehyde or 2% paraformaldehyde + 0.15% picric acid in 0.1 M phosphate buffer, then transferred to the buffer. After 24-48 hours, the kidneys were sectioned with a Vibratome at 200 or 300 micron and incubated in the primary antiserum for 18 hours at room temperature. A sparse plexus of VIPI nerves innervates the rat renal calyx. Some VIPI nerves innervate interlobar arteries and each succeeding segment of the arterial tree including afferent arterioles, but most innervate arcuate and interlobular arteries. VIPI axons do not innervate each arcuate artery or each interlobular branch of an arcuate artery with equal density. Although some axons follow each interlobular branch, most form a dense plexus on only one or two branches. Therefore, most VIPI nerves in the rat kidney innervate a restricted segment of the renal arterial tree. These nerves may be efferent and may selectively dilate arcuate and smaller arteries, or they may be afferent and may sense local changes in mechanical or chemical parameters.
Anat Rec 1987 Oct
PMID:Vasoactive intestinal peptide-immunoreactive nerves in the rat kidney. 368 73

The aim of this immunohistologic study was to determine how lymphocytes and accessory cells associated with antigen processing are distributed in the domes of Peyer's patches. Monoclonal antibodies against mouse B cells (anti-B220), T cells (anti-Thy-1.2), T helper cells (anti-L3T4), T cytotoxic/suppressor cells (anti-Lyt-2), macrophages (anti-Mac-1), and Ia+ cells (anti-I-Ad) were applied to cryostat sections of mouse Peyer's patches and visualized with peroxidase-conjugated avidin-biotin complexes (ABC). The subepithelial dome, the dome epithelium, and the follicle crypt epithelium were ech surveyed for cells labeled with these antibodies. Each region had a characteristic and nonrandom distribution of labeled cells. In the subepithelial dome, B cells, T helper cells, macrophages, and Ia+ accessory cells formed a cellular meshwork between follicle and the dome epithelium. T cytotoxic/suppressor cells were sparse beneath the epithelium. In the dome epithelium, solitary and paired lymphocytes occurred both above and below the level of epithelial cell nuclei. B cells predominated, and both T helper cells and T cytotoxic/suppressor cells were present. B cells sometimes aggregated in small clusters. Macrophages were rarely observed in the epithelium. The distribution of cells in the dome epithelium was distinctly different from non-Peyer's patch intestine where T cytotoxic/suppressor cells predominated and B cells were few in number. Although lymphocytes were usually absent in crypts outside Peyer's patches, in follicle crypts B cells and T cells were seen but not Ia+ accessory cells or macrophages. Most of the T cells in the crypt epithelium were T cytotoxic/suppressor cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1986 Jun
PMID:Differential distribution of lymphocytes and accessory cells in mouse Peyer's patches. 372 11


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