Immunocytochemistry is a laboratory technique used to visualize the presence and location of specific proteins or antigens in horse tissue or cell samples. This method employs antibodies that bind to target proteins, which are then detected using various markers, such as fluorescent dyes or enzymes. In equine research, immunocytochemistry is applied to study cellular processes, disease mechanisms, and tissue pathology. It allows researchers to observe the distribution and expression of proteins within horse tissues, aiding in the understanding of equine diseases and conditions. This page compiles peer-reviewed research studies and scholarly articles that explore the application, methodology, and findings of immunocytochemistry in equine science.
Gamboa S, Ramalho-Santos J.Proteins implicated in the "SNARE hypothesis" for membrane fusion have been characterized in the acrosome of several mammalian species, and a functional role for these proteins during the acrosome reaction has been proposed. We have investigated the presence of SNAREs in equine sperm, using semen samples obtained from stallions with varying fertility. Immunocytochemical analysis revealed that members of different SNARE families can be detected on the acrosome of equine sperm, notably in the acrosomal cap and equatorial segment. These proteins include the t-SNARE syntaxin, the v-SNARE synaptobr...
Hess MF, Roser JF.The large amount of testicular estrogens produced by the stallion is unique compared to the amounts found in other domestic species. Although the cellular locale of the cytochrome P450 aromatase (P450arom) enzyme that converts C19 androgens to C18 estrogens has been identified in the Leydig cell of adult equine testis, the location in the immature equine testis is not known. The goal of this work was to localize the enzyme in colts and stallions during sexual development. Testes were obtained from prepubertal (n=7), pubertal (n=6), and postpubertal (n=8) colts and stallions during both the bre...
Robertson SA, Andrew SE.To determine if opioid growth factor (OGF, [Met5]enkephalin) and its specific receptor (OGFr) are present in normal cat, dog and horse cornea. Methods: Normal dog, cat and horse. Methods: Corneas were obtained from animals euthanized for reasons unrelated to this project. One cornea from each of three normal cats, dogs and horses was evaluated. The right or left cornea from each animal was chosen randomly. Corneas were harvested and placed in corneal storage media for transport to The M.S. Hershey Medical Center of The Pennsylvania State University where immunocytochemistry techniques were use...
Gläser KE, Davies ME, Jeffcott LB.This study was designed to examine a new role for cysteine proteinases in the process of endochondral ossification. Objective: The aim of the present study was to investigate the presence and distribution of cathepsin B and cathepsin L in equine articular cartilage during development. Methods: Full-depth cartilage samples from a total of 40 horses (age range: 4 month fetuses to 2 years) were examined and enzymes detected by immunocytochemical localisation. Results: Observations on the presence of cathepsins B and L revealed significant age-related differences, resulting in clear division of th...
Tremoleda JL, Schoevers EJ, Stout TA, Colenbrander B, Bevers MM.Meiotic maturation of mammalian oocytes is a complex process during which microfilaments and microtubules provide the framework for chromosomal reorganisation and cell division. The aim of this study was to use fluorescence and confocal laser scanning microscopy to examine changes in the distribution of these important cytoskeletal elements and their relationship to chromatin configuration during the maturation of horse oocytes in vitro. Oocytes were cultured in M199 supplemented with pFSH and eLH and, at 0, 12, 24, and 36 hr after the onset of culture, they were fixed for immunocytochemistry ...
Raulo SM, Sorsa TA, Kiili MT, Maisi PS.To determine collagenase activity and evaluate matrix metalloproteinase (MMP)-8 and MMP-13 in horses with chronic obstructive pulmonary disease (COPD). Methods: 12 horses with COPD and 12 healthy control horses. Methods: Collagenase activity was determined by use of an assay for degradation of type-I collagen. Western immunoblot analysis was used to identify interstitial collagenases MMP-8 and MMP-13 in tracheal epithelial lining fluid (TELF). Immunocytochemistry and in situ hybridization were used to determine cellular expression of these 2 collagenases in cells in bronchoalveolar lavage flui...
Raulo SM, Sorsa T, Tervahartiala T, Pirilä E, Maisi P.Gelatinolytic activity was analysed to study whether elevated activity previously found at the tracheal level of the respiratory tract of horses with chronic obstructive pulmonary disease (COPD) could also be found at the lower part of the respiratory tract. Furthermore, presence and significance of the gelatinolytic matrix metalloproteinases (MMPs) MMP-2 and MMP-9 in respiratory secretions of healthy and COPD horses were determined. Elevated gelatinolytic matrix metalloproteinases were detected in bronchoalveolar and tracheobronchial secretions from COPD horses. The main pathologically elevat...
Wooding FB, Morgan G, Fowden AL, Allen WR.The placenta is the only channel for transport of nutrients to the conceptus and the fetal nutrient demands increase exponentially to term. The 9 kDa calcium binding protein (calbindin, 9CBP) and the iron binding protein uteroferrin (UF) are proving to be reliable markers for epithelia that mediate active transcellular calcium and iron transport and the glucose transporter proteins (GT1 and GT3) for glucose transport by facilitated diffusion. Light and electron microscope immunocytochemistry have been used on perfusion fixed resin embedded material to establish the distribution of 9CBP, UF, GT...
Walters KW, Corbin CJ, Anderson GB, Roser JF, Conley AJ.Estrogen production by the preimplantation equine embryo is presumed to be important in maternal-conceptus communication in the mare. The synthesis of C(18) estrogens from C(19) androgens requires cytochrome P450 aromatase (P450(arom)) in the conceptus, but little information is available on the specific tissue location or potential developmental patterns of expression for the horse. The goal of this research was to localize P450(arom) in the equine conceptus by immunocytochemistry and in situ hybridization. Intact blastocyst-stage embryos were collected by nonsurgical flush on Days 12-15 of p...
Siedek EM, Honnah-Symns N, Fincham SC, Mayall S, Hamblin AS.Monoclonal antibodies (mAbs) recognizing equine macrophages are scarce. The present study compared the immunocytochemical staining of various equine tissues (lymphoid tissue, lung, liver, small intestine, skin and blood leucocytes) by an antibody, Ki-M6, which detects CD68 in human macrophages and dendritic cells, and by a new anti-equine mAb, JB10, with staining produced by two previously described anti-equine macrophage mAbs, CZ2.2 and CZ3.3. Ki-M6 was shown to identify equine macrophages, which had a distribution different from those identified by CZ2.2 and CZ3.3. JB10 identified equine mac...
Cornelissen BP, Buma P, Rijkenhuizen AB, Barneveld A.To localize and determine relative frequency of occurrence of calcitonin gene-related peptide (CGRP) and substance P (SP) fibers in the proximal sesamoid bones (PSB) and adjacent structures in sound horses. Methods: 4 foals and 3 adult horses. Methods: Medial PSB and adjacent ligaments of both forelimbs were collected, flushed, and fixed in buffered 4% formalin. The left PSB were cut into 5 longitudinal, sagittally oriented slabs, and the right PSB were cut into 5 transverse slabs. After decalcification in EDTA, slices were transferred to a 30% sucrose solution, deep frozen, sectioned (80 micr...
Day WE, Bowen JA, Barhoumi R, Bazer FW, Burghardt RC.This investigation examines the relationship between implantation strategy and gap junction protein expression in uterine endometrium. The pattern of gap junction and connexin protein expression was analyzed in porcine and equine endometrium from cycling and pregnant animals using electron microscopy and immunocytochemistry. Functional analysis of cell-cell communication was also monitored by laser cytometry in primary cultures of endometrial epithelial cells. Gap junctions were detected in endometrial stroma of cycling and pregnant animals, which was correlated with immunoreactive Cx43 within...
Rahmanian MS, Thompson DL, Melrose PA.Gonadotropin-specific primary antisera and gold-conjugated secondary antibodies were used to immunocytochemically localize gonadotropins in the anterior pituitary of intact pony mares. Electron microscopy was then used to characterize the ultrastructure and immunoreactive staining characteristics of equine gonadotropes. Cells containing LH were morphologically indistinguishable from those containing FSH. Gonadotropes were relatively large and commonly had eccentric nuclei. The rough endoplasmic reticulum was well developed and dilated. Secretory granules were present in two morphologically dis...
Tan JH, Nanbo Y, Oikawa M, Kiso Y, Sasaki F.To analyze the sex difference in 6 kinds of adenohypophyseal cells of Mongolian ponies and the effect of prepubertal orchidectomy on adenohypophyseal cells. Methods: Pituitary glands collected from 15 adult Mongolian ponies, 5 to 10 years old: 5 stallions, 5 mares, and 5 geldings, orchidectomized between the ages of 1 and 2 years. Methods: Morphologic comparison of 6 kinds of adenohypophyseal cells among mares, stallions, and geldings was done, using immunocytochemistry and morphometry. Results: A sex difference was evident in the percentage of somatotrophs, gonadotrophs (follicle-stimulating ...
Rahmanian MS, Thompson DL, Melrose PA.The ultrastructural and immunoreactive staining characteristics of cells containing prolactin (lactotropes) and growth hormone (GH; somatotropes) in the anterior pituitaries of gonadally intact pony mares were studied at the electron microscopic level. Lactotropes included two morphological subsets: Type I cells were larger and contained large, dense, polymorphic granules that were scattered throughout the cytoplasm; Type II cells were smaller and contained small, dense, polymorphic granules that were predominantly found in peripheral areas of the cytoplasm. Lactotropes constituted 5 to 16% of...
Chao TI, Grosche J, Friedrich KJ, Biedermann B, Francke M, Pannicke T, Reichelt W, Wulst M, Mühle C, Pritz-Hohmeier S, Kuhrt H, Faude F, Drommer W....Müller cells from 22 mammalian species were subjected to morphological and electrophysiological studies. In the 'midperiphery' of retinae immunocytochemically labeled for vimentin, estimates of Müller cell densities per unit retinal surface area, and of neuron-to-(Müller) glia indices were performed. Müller cell densities were strikingly similar among the species studied (around 8000-11,000 mm-2) with the extremes of the horse ( or = 20,000 mm-2). By contrast, the number of neurons per Müller cell varied widely, being clustered at 6-8 (in retinae with many cones), at about 16, and at up t...
Hernandez-Vidal G, Jeffcott LB, Davies ME.The distribution of cathepsin D in normal equine growth cartilage has been examined immunocytochemically using an antiserum raised against human cathepsin D. The cross-reactivity and specificity of the antiserum for equine cathepsin D was confirmed, and its lysosomal localisation was demonstrated in horse skin fibroblasts by confocal scanning microscopy. Cultured horse chondrocytes were heterogenous in their expression of cathepsin D. Heterogeneity of distribution of the enzyme was also seen in chondrocytes in cartilage from different anatomical sites. A high level of cathepsin D was observed ...
Klonisch T, Mathias S, Cambridge G, Hombach-Klonisch S, Ryan PL, Allen WR.In situ hybridization employing a cRNA probe derived from a 428-bp fragment of equine relaxin was used to localize relaxin mRNA, and immunocytochemistry was used to localize relaxin itself, in tissues of the placenta-endometrium interface recovered between 33 and 153 days of gestation from mares carrying intraspecific horse, interspecific mule and extraspecific donkey conceptuses. Immunocytochemical staining was also used to localize trophoblast-specific and class I major histocompatibility complex (MHC) antigens on some specimens. Relaxin mRNA and relaxin were both present in the single-cell ...
Sandmann D, Boycott BB, Peichl L.The morphology of horizontal cells chiefly of the horse, but also of asses, mules, and a zebra, has been examined by Lucifer yellow injections into lightly fixed retinae and by immunocytochemistry. In common with other mammals, equids have a B-type horizontal cell, i.e., a cell with dendrites synapsing with cones and possessing a single axon synapsing with rods. Most mammalian retinae have a further type of horizontal cell, the A-type, also synapsing with cones but without an axon. The second type of horizontal cell in equids also has no axon; otherwise, it is most unusual. Compared with other...
Thomas PG, Ignotz GG, Ball BA, Miller PG, Brinsko SP, Currie B.Oviduct epithelial cells (OEC) increasingly are used to support embryonic development and to study gamete interactions with the female reproductive tract in vitro. This series of experiments was designed to characterize monolayers derived from oviduct epithelium. Epithelial cells harvested from the isthmus and ampulla of the oviducts of five estrous mares were cultured with or without the basal lamina extract, Matrigel. Within each group OEC were cultured in the presence of either estradiol-17 beta or a carrier control. All groups were subcultured three times. Epithelial cell morphology and fu...
Grandi D.An ultrastructural and immunohistochemical analysis was performed on the epiphysis of two species of mammals: mink and horse. These animals present a reproductive cycle that varies along the year according to the light exposition. The aim of the study was to define possible structural changes connected with the gonadal activity. Structural aspects in common emerged such as those in concern with the epiphysis cells, their immunocytochemical characteristics (NSE and Synaptophysin positivity of pinealocytes, as well as CFAP positivity of further cells), and those in concern with the general ultra...
Eisenhauer KM, McCue PM, Nayden DK, Osawa Y, Roser JF.Stallion testes secrete large amounts of estrogens, but the cellular location of the enzyme that converts androgens to estrogens, cytochrome P450 aromatase, has not been determined. The goal of the present study was to immunocytochemically localize stallion testicular aromatase using a polyclonal antibody generated against human placental cytochrome P450 aromatase. Testes were obtained from 12 stallions from 2 to 23 years of age, during both the breeding and non-breeding seasons. Immunoreactivity was confined to the Leydig cells in all testes examined. No immunostaining was observed in the Ser...
Bowker RM, Rockershouser SJ, Linder K, Vex KB, Sonea IM, Caron JP.The innervation of the navicular bone (os sesamoideum distale) and its suspensory ligaments (ligamenta sesamoidea collateralia) (CSL) or proximal suspensory ligament and the ligamentum sesamoideum distale impar or the distal sesamoidean impar ligament (DS-impar ligament) was examined using combined anatomical techniques of silver impregnation and immunocytochemistry. Silver impregnation studies revealed an abundance of nerve fibres present in both the CSL and DS-impar ligament with the latter having relatively more nerve fibres. These silver-impregnated nerves coursed parallel to and were asso...
Bowker RM, Sonea IM, Vex KB, Caron JP.Substance P (SP) immunocytochemistry and receptor autoradiography were used to define the innervation of the equine synovial membrane of joints equivalent to the wrist and knuckle of man. SP-immunoreactive fibers were mainly concentrated around blood vessels in the subsynovial layer, although not exclusively, while in the more distal joint, SP fibers were more frequently seen in the synovial surface layer. Iodinated SP receptor autoradiography studies revealed silver grain concentrations in the advential layer of blood vessels associated with the vasa vasorum, on the vascular endothelium and i...
Bowker RM, Rockershouser SJ, Vex KB, Sonea IM, Caron JP, Kotyk R.To determine whether the distal interphalangeal (DIP) joint directly or indirectly communicates with the navicular bursa (bursa podotrochlearis) and to identify sensory nerves in these synovial structures that might be desensitized by intra-articular injections of anesthetics, Evans blue dye in physiologic saline solution, Luxol fast blue dye with mepivicaine, or commercial latex was injected into the DIP joint (5 ml) or the navicular bursa (3 ml) of 152 digits obtained from horses or ponies at necropsy. The digits were frozen, cut with a band saw, and examined for distribution of dye or latex...
Bowker RM, Brewer AM, Vex KB, Guida LA, Linder KE, Sonea IM, Stinson AW.Two types of sensory receptors were located in the equine foot, using anatomic techniques. Histologic examination of stained hoof sections revealed lamellated corpuscles in the hoof dermis, which had many of the morphologic characteristics of Pacinian corpuscles. These sensory receptors were restricted to the palmar (caudal) aspects of the solar dermis of the heel. A second type of receptor was detected by use of immunocytochemistry, indicating apparently naked nerve endings containing the neuropeptide calcitonin gene-related peptide-like immunoreactivity in skin, solar dermal tubules, and the...
Bowker RM, Abhold RH, Caron JP, Sonea IM, Vex KB, Kotyk R.Immunocytochemical analysis of equine synovial membranes revealed presence of several neuropeptides, including substance P (SP), neurokinin A, and neuropeptide Y, in nerves of the radiocarpal, middle carpal, and metacarpophalangeal (fetlock) joints. Within the subsynovium, these neuropeptides were located perivascularly, whereas in the fronds, only neuropeptide Y was restricted to the vessels of the synovial membrane. Only SP and neurokinin A were found in the intimal layer. The intimal layer of the metacarpophalangeal joint contained more SP-immunoreactive fibers than were observed in the int...
Griffiths IR, Kyriakides E, Smith S, Howie F, Deary AW.Equine grass sickness (EGS) is a primary dysautonomia characterised pathologically by lesions in autonomic ganglia, enteric plexi and specific nuclei in the CNS. Immunocytochemistry and lectin histochemistry of the autonomic ganglia were used to determine whether abnormalities can be detected in specific proteins or cellular organelles. EGS ganglia contained a mixture of morphologically normal and abnormal neurons, the former appearing identical to cells from control animals. Affected cells showed marked disturbances in neurofilament (NF) proteins and beta-tubulin, major components of the cyto...
Bakst M, Hadick S, Proudman J, Maruyama K.This study was conducted to determine the crossreactivity of antisera to human prolactin (PRL), adrenocorticotropic hormone (ACTH), and growth hormone (GH) to turkey pituicytes. In addition, crossreactivities of the above antisera and antiserum to turkey GH to pituicytes of turkey, cat, rabbit, horse, owl monkey, and human were evaluated. Results of the immunocytochemical localizations showed that with one exception antisera to human hormones were positive for each species tested. Turkey pituicytes failed to crossreact with antiserum to human GH. Likewise, antiserum to turkey GH failed to cros...
Sinha AK, Rose RJ, Pozgaj I, Hoh JF.The histochemical ATPase method for muscle fibre typing was first described by Brooke and Kaiser in 1970. However, problems have been found with the subdivision of type II fibres using this technique. To determine whether indirect myosin immunocytochemistry using anti-slow (5-4D), anti-fast (1A10) and anti-fast red (5-2B) monoclonal antibodies with cross reactivity for type I, II and IIa fibres, respectively, in a number of species, could identify three fibre types in equine skeletal muscle, data on fibre type composition and fibre size obtained using the two different techniques were compared...
Sabeur K, Ball BA.Carbohydrate-binding proteins are thought to be involved in a myriad of sperm functions including sperm-oviductal and sperm-zona interactions. Recent studies in our laboratory have characterized galactose-binding proteins on equine spermatozoa as possible candidate molecules for sperm adhesion to oviduct epithelial cells. In the current study, equine sperm membrane proteins were subjected to galactose-affinity chromatography, and bound proteins were eluted with excess galactose in a calcium-free buffer. The eluted fraction recovered after galactose-affinity chromatography was used for generati...
Griffiths IR, Kyriakides E, Smith S, Howie F, Deary AW.Equine grass sickness (EGS) is a primary dysautonomia characterised pathologically by lesions in autonomic ganglia, enteric plexi and specific nuclei in the CNS. Immunocytochemistry and lectin histochemistry of the autonomic ganglia were used to determine whether abnormalities can be detected in specific proteins or cellular organelles. EGS ganglia contained a mixture of morphologically normal and abnormal neurons, the former appearing identical to cells from control animals. Affected cells showed marked disturbances in neurofilament (NF) proteins and beta-tubulin, major components of the cyto...
Yang Q, Pinto VMR, Duan W, Paxton EE, Dessauer JH, Ryan W, Lopez MJ.Damage to an ectodermal-mesodermal interface like that in the equine hoof and human finger nail bed can permanently alter tissue structure and associated function. The purpose of this study was to establish and validate culture of primary progenitor cell isolates from the ectodermal-mesodermal tissue junction in equine hooves, the stratum internum, with and without chronic inflammation known to contribute to lifelong tissue defects. The following were evaluated in hoof stratum internum cell isolates up to 5 cell passages (P): expansion capacity by cell doublings and doubling time; plasticity ...
Soverchia L, Mosconi G, Ruggeri B, Ballarini P, Catone G, Degl'Innocenti S, Nabissi M, Polzonetti-Magni AM.Proopiomelanocortin (POMC) is a precursor protein that contains the sequences of several bioactive peptides including adrenocorticotropin (ACTH), beta-endorphin (beta-EP), and melanocyte-stimulating-hormone (MSH). POMC is synthesized in the pituitary gland, brain, and many peripheral tissues. Immunoreactive POMC-derived peptides as well as POMC-like mRNA have been evidenced in several nonpituitary tissues, thus suggesting that POMC is actively synthesized by these tissues. The present study was aimed at evaluating if also in the case of stallion POMC-derived peptide, beta-EP, is produced local...
Siedek EM, Honnah-Symns N, Fincham SC, Mayall S, Hamblin AS.Monoclonal antibodies (mAbs) recognizing equine macrophages are scarce. The present study compared the immunocytochemical staining of various equine tissues (lymphoid tissue, lung, liver, small intestine, skin and blood leucocytes) by an antibody, Ki-M6, which detects CD68 in human macrophages and dendritic cells, and by a new anti-equine mAb, JB10, with staining produced by two previously described anti-equine macrophage mAbs, CZ2.2 and CZ3.3. Ki-M6 was shown to identify equine macrophages, which had a distribution different from those identified by CZ2.2 and CZ3.3. JB10 identified equine mac...
Chan G, Morgan WH, Yu DY, Balaratnasingam C.Astrocytes are critical for the maintenance of retinal ganglion cell (RGC) axonal function and viability, and form a key component of the functional neurovascular unit. Recently, we described the quantitative properties of astrocytes in relation to the capillary distributions in optic nerve laminar regions. Here, we provide a quantitative analysis of astrocytes and RGC axons in longitudinal sections of optic nerve tissue. Histological and immunocytochemical techniques are used to demonstrate the density of astrocytes, RGC axons and glia-neuron ratios across the pre laminar, lamina cribrosa and...
Rahmanian MS, Thompson DL, Melrose PA.Gonadotropin-specific primary antisera and gold-conjugated secondary antibodies were used to immunocytochemically localize gonadotropins in the anterior pituitary of intact pony mares. Electron microscopy was then used to characterize the ultrastructure and immunoreactive staining characteristics of equine gonadotropes. Cells containing LH were morphologically indistinguishable from those containing FSH. Gonadotropes were relatively large and commonly had eccentric nuclei. The rough endoplasmic reticulum was well developed and dilated. Secretory granules were present in two morphologically dis...
Tarko A, Fabová Z, Kotwica J, Valocký I, Alrezaki A, Alwasel S, Harrath AH, Sirotkin AV.The influence of environmental contaminant toluene and of plant fennel (Foeniculum vulgare Mill.) on reproduction are reported, but the mechanisms of their action and the protective effect of fennel on contaminant influence remain to be elucidated. In this study, we hypothesized that toluene and fennel directly affects basic ovarian cell functions, and that fennel can be used as an appropriate natural protective agent against the potential adverse effects of toluene. This study aimed to examine the action of toluene (20 μg/mL) and fennel extract (0, 1, 10, 100 μg/mL), and assess their comb...
Bishop AE, Hodson NP, Major JH, Probert L, Yeats J, Edwards GB, Wright JA, Bloom SR, Polak JM.In recent years, distinct changes in regulatory peptides have been found in a number of gastrointestinal diseases. Grass sickness is a fatal disease of horses for which the etiology has yet to be fully ascertained. In this study, the peptide-containing nerves and ganglionic and mucosal endocrine cells of the ileum, colon and rectum were investigated in horses with sub-acute or chronic grass sickness and compared with normal controls using immunocytochemistry, at both the light and electron microscopical levels, and radioimmunoassay. A substantial loss of both peptide-containing cells and nerve...
Cummings JF, Sellers AF, Lowe JE.The distribution of the putative motor excitatory neurotransmitter, substance P, was studied immunocytochemically in the left dorsal colon of four normal control ponies and three ponies with amitraz-induced impaction colic. Substance P-like immunoreactivity in the control ponies was observed in nerve fibres in all layers of the bowel wall and in the nerve cell bodies of the enteric ganglia. The substance P-like immunoreactivity was clearly more intense in the cell bodies of submucosal ganglia than in those of the myenteric ganglia. The internodal nerve strands of the myenteric plexus were very...
The measurement of cell proliferation and cell viability using 5'bromo-2'deoxy-uridine (BrdU) labelling has been described in several cell types and species. The aim of this study was to adapt this technique to equine embryos and to compare the index of DNA replication (S-phase) between equine and caprine embryos. Seventeen equine embryos were recovered at day 6.5 post-ovulation and 20 caprine embryos were recovered at day 7 after the onset of estrus. Equine embryos were incubated during 1h at 39 degrees C in PBS containing 1mM of BrdU. Embryos were then treated in 0.05% trypsin during 15 min ...
Cozzi B.A combined ultrastructural and immunocytochemical study was performed on the pineal gland of the horse in order to identify the cell types present and describe their characteristics. Comparisons have been made with other mammals. Two main cell types are present: pinealocytes and glial cells. Pinealocytes display different degrees of electron density in the nucleus and the cytoplasm, yet no ultrastructural feature supports the idea of separate populations. Putative secretory materials are stored in vesicles related to the Golgi apparatus. A variety of electron-dense bodies are present in the cy...
Sinha AK, Rose RJ, Pozgaj I, Hoh JF.The histochemical ATPase method for muscle fibre typing was first described by Brooke and Kaiser in 1970. However, problems have been found with the subdivision of type II fibres using this technique. To determine whether indirect myosin immunocytochemistry using anti-slow (5-4D), anti-fast (1A10) and anti-fast red (5-2B) monoclonal antibodies with cross reactivity for type I, II and IIa fibres, respectively, in a number of species, could identify three fibre types in equine skeletal muscle, data on fibre type composition and fibre size obtained using the two different techniques were compared...
Furuoka H, Shirakawa T, Taniyama H, Ohishi H, Satoh H, Itakura C.Pancreatic tissue from 20 horses was examined using immunocytochemical techniques. In aged horses, neogenesis of endocrine cells, neoformation, and hyperplasia of islets occurred closely associated with the pancreatic duct; these changes were regarded as nesidioblastosis. In addition, pancreatic fibrosis accompanied by ductal proliferation and endocrine neogenesis was considered a regenerative change. Thus, the origin of neoformation in the endocrine pancreas was in the ductal system, and it is suggested that the pancreatic endocrine cells were of endodermal origin.
Klonisch T, Mathias S, Cambridge G, Hombach-Klonisch S, Ryan PL, Allen WR.In situ hybridization employing a cRNA probe derived from a 428-bp fragment of equine relaxin was used to localize relaxin mRNA, and immunocytochemistry was used to localize relaxin itself, in tissues of the placenta-endometrium interface recovered between 33 and 153 days of gestation from mares carrying intraspecific horse, interspecific mule and extraspecific donkey conceptuses. Immunocytochemical staining was also used to localize trophoblast-specific and class I major histocompatibility complex (MHC) antigens on some specimens. Relaxin mRNA and relaxin were both present in the single-cell ...
Cornelissen BP, Buma P, Rijkenhuizen AB, Barneveld A.To localize and determine relative frequency of occurrence of calcitonin gene-related peptide (CGRP) and substance P (SP) fibers in the proximal sesamoid bones (PSB) and adjacent structures in sound horses. Methods: 4 foals and 3 adult horses. Methods: Medial PSB and adjacent ligaments of both forelimbs were collected, flushed, and fixed in buffered 4% formalin. The left PSB were cut into 5 longitudinal, sagittally oriented slabs, and the right PSB were cut into 5 transverse slabs. After decalcification in EDTA, slices were transferred to a 30% sucrose solution, deep frozen, sectioned (80 micr...
Bowker RM, Rockershouser SJ, Linder K, Vex KB, Sonea IM, Caron JP.The innervation of the navicular bone (os sesamoideum distale) and its suspensory ligaments (ligamenta sesamoidea collateralia) (CSL) or proximal suspensory ligament and the ligamentum sesamoideum distale impar or the distal sesamoidean impar ligament (DS-impar ligament) was examined using combined anatomical techniques of silver impregnation and immunocytochemistry. Silver impregnation studies revealed an abundance of nerve fibres present in both the CSL and DS-impar ligament with the latter having relatively more nerve fibres. These silver-impregnated nerves coursed parallel to and were asso...
Yang Q, Lopez MJ.The equine hoof dermal-epidermal interface requires progenitor cells with distinct characteristics. This study was designed to provide accurate ultrastructural depictions of progenitor cells isolated from inflamed tissue and normal tissue before and after cryopreservation and following selection of cells expressing both keratin (K) 14 (ectodermal) and cluster of differentiation (CD) 105 (mesodermal). Passage 3 cell ultrastructure was assessed following 2D culture and after 3D culture on decellularized hoof tissue scaffolds. Outcome measures included light, transmission electron, and scanning e...
Miller L, Woodward EM, Campos JR, Squires EL, Troedsson M.The objectives of this study were to (i) verify localization of SP22 on fresh, cooled, and frozen/thawed equine spermatozoa and to (ii) determine SP22 mRNA and protein expression in equine testicular and epididymal tissues. Immunocytochemistry and Western blots were performed on the spermatozoa samples. Northern blots and Western blots were performed on the tissue samples. The immunocytochemistry revealed the presence of SP22 in all samples tested. The fresh spermatozoa stained predominantly over the equatorial segment as did the samples cooled for 1 and 2 days. The samples cooled for 3 days, ...
Walters KW, Corbin CJ, Anderson GB, Roser JF, Conley AJ.Estrogen production by the preimplantation equine embryo is presumed to be important in maternal-conceptus communication in the mare. The synthesis of C(18) estrogens from C(19) androgens requires cytochrome P450 aromatase (P450(arom)) in the conceptus, but little information is available on the specific tissue location or potential developmental patterns of expression for the horse. The goal of this research was to localize P450(arom) in the equine conceptus by immunocytochemistry and in situ hybridization. Intact blastocyst-stage embryos were collected by nonsurgical flush on Days 12-15 of p...
Gautam A, Dubey JP, Saville WJ, Howe DK.Sarcocystis neurona is a two-host coccidian parasite whose complex life cycle progresses through multiple developmental stages differing at morphological and molecular levels. The S. neurona merozoite surface is covered by multiple, related glycosylphosphatidylinositol-linked proteins, which are orthologous to the surface antigen (SAG)/SAG1-related sequence (SRS) gene family of Toxoplasma gondii. Expression of the SAG/SRS proteins in T. gondii and another related parasite Neospora caninum is life-cycle stage specific and seems necessary for parasite transmission and persistence of infection. I...
Campos-Junior PH, Costa GM, de Avelar GF, Segatelli TM, Lacerda SM, Aponte PM, de França LR.Morphometry is a classical quantitative method often used in biology to provide a data basis for functional interpretations/interactions of a particular organ or system. Herein we took advantage of this valuable approach to evaluate the spermatogonial stem cell niche using the horse testis and immunocytochemical localization of GFRA1 [glial cell line-derived neurotrophic factor receptor produced by Sertoli cells)] as an example. Using the NIH ImageJ free software, we describe in detail all the necessary steps to investigate this specific and crucial microenvironment. Based on several recently ...
Sonmez K, Gurel A, Takai S.The aim of the present study was to develop an immunocytochemical procedure for the early detection and demonstration of Rhodococcus equi in smears of tracheal aspirates taken from live foals in field conditions. Tracheal wash samples were collected from thoroughbred foals, aged 1-5 months and located in studs around Bursa and Istanbul, Turkey. Some foals were suspected of having R. equi infection on the basis of clinical examination (n=56) and others were unaffected control animals (n=54). Serum samples were also collected from each foal for testing for the presence of R. equi-specific antibo...
Duran MC, Starrak G, Dickinson R, Montgomery J.After physical examination, ultrasonographic evaluation of thorax and abdomen, and peritoneal fluid analysis, gastrointestinal neoplasia with suspected diffuse peritoneal metastasis was diagnosed in a 17-year-old Arabian gelding. The owner elected euthanasia and declined postmortem examination. Immunocytochemistry analysis of the peritoneal fluid resulted in a diagnosis of B-cell lymphoma. Immunocytochimie du liquide péritonéal utilisée pour le diagnostic d’un cas possible de lymphome gastro-intestinal à cellules B. Après un examen physique, une évaluation échographique du thorax et d...
Gray PR, Derksen FJ, Robinson NE, Slocombe RF, Peters-Golden ML.We have developed an alternative method for examining equine tracheal epithelial arachidonic acid (AA) metabolism that utilizes strips of pseudostratified columnar epithelium attached to a layer of elastic tissue 80 to 130 microns thick. We compared the responses of this preparation with those of enzymatically dispersed suspensions of tracheal epithelium obtained from the same animal. Strips incubated with [3H]AA incorporated 40.8 +/- 3.6% of added radioactivity and released 2.55 +/- 0.23% of incorporated radioactivity when stimulated with 5 microM A23187. Values for the cell suspension were 5...
Grandi D.An ultrastructural and immunohistochemical analysis was performed on the epiphysis of two species of mammals: mink and horse. These animals present a reproductive cycle that varies along the year according to the light exposition. The aim of the study was to define possible structural changes connected with the gonadal activity. Structural aspects in common emerged such as those in concern with the epiphysis cells, their immunocytochemical characteristics (NSE and Synaptophysin positivity of pinealocytes, as well as CFAP positivity of further cells), and those in concern with the general ultra...
Bowker RM, Rockershouser SJ, Vex KB, Sonea IM, Caron JP, Kotyk R.To determine whether the distal interphalangeal (DIP) joint directly or indirectly communicates with the navicular bursa (bursa podotrochlearis) and to identify sensory nerves in these synovial structures that might be desensitized by intra-articular injections of anesthetics, Evans blue dye in physiologic saline solution, Luxol fast blue dye with mepivicaine, or commercial latex was injected into the DIP joint (5 ml) or the navicular bursa (3 ml) of 152 digits obtained from horses or ponies at necropsy. The digits were frozen, cut with a band saw, and examined for distribution of dye or latex...
Bakst M, Hadick S, Proudman J, Maruyama K.This study was conducted to determine the crossreactivity of antisera to human prolactin (PRL), adrenocorticotropic hormone (ACTH), and growth hormone (GH) to turkey pituicytes. In addition, crossreactivities of the above antisera and antiserum to turkey GH to pituicytes of turkey, cat, rabbit, horse, owl monkey, and human were evaluated. Results of the immunocytochemical localizations showed that with one exception antisera to human hormones were positive for each species tested. Turkey pituicytes failed to crossreact with antiserum to human GH. Likewise, antiserum to turkey GH failed to cros...
Szober CM, Hauck SM, Euler KN, Fröhlich KJ, Alge-Priglinger C, Ueffing M, Deeg CA.The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses' vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, an...
Gläser KE, Davies ME, Jeffcott LB.This study was designed to examine a new role for cysteine proteinases in the process of endochondral ossification. Objective: The aim of the present study was to investigate the presence and distribution of cathepsin B and cathepsin L in equine articular cartilage during development. Methods: Full-depth cartilage samples from a total of 40 horses (age range: 4 month fetuses to 2 years) were examined and enzymes detected by immunocytochemical localisation. Results: Observations on the presence of cathepsins B and L revealed significant age-related differences, resulting in clear division of th...
