Epithelial cells with vacuoles containing 54,000 dalton sialoglycoprotein in the mouse epididymal duct

Epididymal ligation in the mouse induced unusual cells, called peculiar pale epithelial cells (hereafter, pale cells), specifically in the epithelium of the corpus epididymidis (ABE et al., 1982a, b). These pale cells had vacuoles with long microvilli on their inner surface. In this study, we found that the vacuoles of the pale cells occurred in normal mice and contained epididymal specific glycoprotein, sialoglycoprotein of 54,000 dalton (SGP54). This was elucidated by indirect immunofluorescence and avidin biotin complex techniques using monoclonal antibody T21 which specifically recognizes SGP54. These immunoreactive pale cells occurred in the distal caput and proximal corpus of the epididymidis. The relationship between the pale cell and SGP54 is discussed.     

Immunohistochemical studies of the epididymal duct in Egyptian water buffalo (Bubalus bubalis)

Using immunohistochemistry (IHC), this study aimed to evaluate the regional distribution pattern of some biologically active proteins in the epididymis of Egyptian water buffalo and to determine the structural-functional relationships of the different epididymal structures. Wax-embedded sections from different regions of the epididymal duct from adult, clinically healthy, buffalo bulls were used. Primary antibodies against angiotensin converting enzyme (ACE), S-100, galactosyltransferase (GalTase), alpha smooth muscle actin (α-SMA), connexin 43 (Cx43) and vascular endothelial growth factor (VEGF) were used for immunohistochemical studies. The results showed that, in addition to the well-known principal and basal cells, the epididymal epithelium, similar to that of other species, possessed apical cells and intraepithelial leukocytes. IHC showed that, with the exception of VEGF which reacted negatively, all antibodies used displayed variable reactivity in the different epididymal structures. Apical cells expressed a strong reaction with ACE along the entire length of the duct. The principal cells in the caput epididymis exhibited a distinct reactivity with S-100 and GalTase. The peritubular muscular coat displayed a marked immunostaining for α-SMA and for Cx43. In conclusion these findings showed a regional-specific distribution pattern, distinct from that in bovine bulls. Some potential functional capacities, especially absorptive and secretory ones, are discussed in relation to the different epididymal regions.     

Immunohistochemical localization of renin in mouse brain

Immunohistochemical studies of mouse brain by the peroxidase-antiperoxidase method using monospecific antimouse renin antibodies has revealed the intracellular localization of renin in stellate and small ovoid cells. Renin-containing ovoid cells were observed in the spinal cord, medulla oblongata, pons, granular layers of the cerebellum, deep cerebellar region, and the lamina terminalis; whereas immunostainable stellate cells were found in the cerebral cortex, hippocampus, dentate gyrus and septum. Intracellular localization of renin rather than intravascular localization supports an endogenous origin of this enzyme in the brain. Wide distribution in different types of cells suggests different types of regulatory mechanisms. Double immunostaining with antigalactocerebroside and antirenin antibodies indicates the presence of renin in oligodendrocytes.     

Immunohistochemical localization of connexin 43 in the developing tooth germ of rat

Distribution of gap junction protein in maxillary tooth germs of 1-day-old rats was examined by immunohistochemistry, using an affinity-purified antibody specific to residues 360–376 of rat connexin (CX) 43. In 1-day-old rats, the maxillary second molar formed the shape of the cusp, but neither dentine nor enamel was formed between the cells of the dental papilla and the inner enamel epithelium. In the tooth germ, CX 43 was expressed in the cells of the stratum intermedium and the inner enamel epithelium. Labelling in the stratum inter-medium was extensive and showed an increasing gradient from peripheral to cuspal regions. CX 43 detected in the inner enamel epithelium was at cell surfaces facing the interface between the dental papilla and the inner enamel epithelium. The cells of the dental papilla and the inner enamel epithelium began differentiation as odontoblasts and secretory ameloblasts respectively, in the cusps of the first molars, where predentine and dentine were formed but enamel matrix was not secreted. CX 43 was present in the stratum intermedium, inner enamel epithelium, preodontoblasts, odontoblasts and subodontoblasts. The incisors showed the most advanced stage of development, where the enamel matrix and calcified dentine were formed in the labial part of the teeth. The CX 43 epitope was seen in the stratum intermedium, inner enamel epithelium, preameloblasts, preodontoblasts, odontoblasts, and subodontoblasts. Immunolabelling was more extensive in the stratum intermedium and subodontoblasts than in preameloblasts, preodontoblasts, and odontoblasts. The immunolabelling in preameloblasts and preodontoblasts was accumulated at cell surfaces facing the predentine. Further, the labelling in preameloblasts and preodontoblasts disappeared or was reduced at the initiation of enamel matrix secretion and calcification of dentine matrix.The present results suggest that gap junctional cell communication has important roles in tooth development. Further, the extensive CX 43 expression in the stratum intermedium and the subodontoblast layer suggests that gap junctions have an important role in amelogenesis and dentinogenesis.     

Cutaneous leishmaniasis in mast cell-deficient W/Wv mice.

Genetically mast cell-deficient W/Wv mice infected with Leishmania amazonensis showed progressive development of ulcerative skin lesions. However, no significant differences between the W/Wv mice and the normal littermates with respect to size of the lesions, anti-Leishmania immunoglobulin E antibody, and the number of eosinophils accumulated in the lesions were observed.     

Immunohistochemical localization of osteoblast activating peptide in the mouse kidney

Osteoblast activating peptide (OBAP) is a newly discovered peptide detected in the rat stomach, which has a major role in osteogenesis. Recently, we revealed its localization in the parietal cells of the rat stomach. There have been no data regarding OBAP expression in the kidney, despite its role in calcium reabsorption in renal tubules. The current study aimed to inspect the expression of OBAP in the kidney of twelve 10-week-old male C3H/HeNJc1 mice using immunohistochemistry, and immunoelectron microscopic localization. The immunohistochemical investigation revealed an OBAP positive reaction mainly in the medulla, which was stronger than the cortex of the kidney and was concentrated in the distal convoluted tubules (DCT), connecting tubules (CT), and the thick limbs of the loop of Henle (HL). Moreover, we clarified that the OBAP was co-distributed with ghrelin and calbindin (markers of the DCT). Interestingly, immunoelectron microscopy demonstrated that OBAP was concentrated in the mitochondrial inner membrane of the DCT and CT. Based on these results, it was concluded that the mitochondria of the DCT, CT, and HL of the mice kidney generate OBAP. Furthermore, our results suggest that OBAP might have a role in the regulation of calcium reabsorption by the renal tubule; however, further investigations are required to clarify this potential role.     

Immunohistochemical localization of HCG and its subunits in testicular germ cell tumours

Summary Native hCG and its and subunits have been localized by immunocytochemistry in 45 testicular germ cell tumours of the testis.Positivity was found for the three molecules in areas with trophoblastic differentiation or in syncytial-like giant cells present in some seminomas. Isolated positivity of hCG was demonstrated in isolated cells usually found in areas of entodermal differentiation of immature malignant teratomas, and probably of neuro-endocrine function.This finding points to genomic derepression in tumour cells and probably also indicates a variability in subunit synthesis and a defect in subunit recombination.Supported by GRANT n^o 3.4530.81 of FRSM and by the FONDS CANCEROLOGIQUE of C.G.E.R.     

Changes in distribution and staining reactivity of PAS-positive material in the mouse epididymal duct after efferent duct ligation

The distribution of spermatozoa and that as well as the staining reaction of PAS-positive material in the epididymal duct were histologically observed in 60 day-old mice and mice 12 h, 24 h, 2 days, and 7 days after efferent duct ligation at 60 days of age. We divided the mouse epididymal duct into five segments; Segments I, II, and III making up the head of the epididymis, Segment IV the body, and Segment V the tail. The cytoplasm of the principal cells in Segment II was PAS-positive. The lumen of the epididymal duct contained spermatozoa and PAS-positive material in the distal portion of Segment I and other segments distal to Segment I. The lumen in Segments IV and V was distended with abundant spermatozoa and PAS-positive material. After efferent duct ligation, spermatozoa disappeared from the lumen in Segments I, II, and III within 2 days, whereas they took 5 days to pass from Segment IV. As spermatozoa disappeared, the luminal material increased in intensity of the PAS-reaction in the distal portion of Segment I and initial portion of Segment II and Segment IV. Segment I became similar in appearance to Segment II 7 days after the ligation. The findings suggest that the PAS-positive material is produced in the distal portion of Segment I and Segment II and accumulates, in the absence of spermatozoa, at the production site and Segment IV, a storage site of spermatozoa; furthermore, Segment I has the capacity of exerting a Segment II-like function, which becomes distinct after blocking the entrance of the testicular fluid.     

Immune response potential of mast cell-deficient W/Wv mice

Immune responses of mast cell-deficient WBB6F1-W/Wv mice and their mast cell-sufficient littermates (LM: WBB6F1-W/+, Wv/+ and +/+) were compared. After a single intravenous injection of sheep erythrocytes (SE), polyvinylpyrrolidone or bacterial lipopolysaccharide, the antigen-specific IgM plaque-forming cell (PFC) response of W/Wv mice was similar to or greater than the response of LM mice. When both primary and secondary injections of SE or chicken gamma-globulin were given to mice and antigen-specific IgG PFC responses quantified, the response of W/Wv again was similar to or greater than that of LM mice. Serum titers of antigen-specific IgE were higher in W/Wv than in LM mice after injections of ovalbumin in alum or infections of Nippostrongylus brasiliensis. Ovalbumin-sensitized W/Wv and LM mice developed active systemic anaphylaxis after ovalbumin challenge. The ability of W/Wv mice to be sensitized for and elicit contact sensitivity (CS) reactions was studied using picryl chloride or dinitrofluorobenzene as sensitizing and challenge agents and quantifying 24-hour reactions by change in ear thickness. SE or methylated bovine serum albumin was used to sensitize and challenge mice for delayed-type hypersensitivity (DTH) reactions which were quantified at 24 h by change in foot pad or ear thickness. CS and DTH reactions of W/Wv and LM mice were similar. No evidence of immune deficiency of W/Wv mice was found.     

Susceptibility of W/Wv mice to murine cytomegalovirus infection

Summary The susceptibility of congenitally anemic W/W^v mice to infection with murine cytomegalovirus (MCMV) was examined. W/W^v mice showed a higher mortality rate and shorter survival time after MCMV infection than did their +/+ littermate mice. In addition, W/W^v mice showed a lower plaque-forming unit (PFU) per 50% lethal dose (LD₅₀) and produced higher titers of infectious virus in various organs. The mortality rate and survival time of W/W^v mice which received a bone marrow graft 4 weeks before infection was completely restored to the level for +/+ mice, suggesting the importance of the cells of myeloid origin.Although natural killer (NK) activity of W/W^v mice was comparable to that of +/+ mice before infection, marked reduction was observed after MCMV infection. Furthermore, OK-432 treatment failed to enhance NK activity of W/W^v mice. Impaired NK response was also completely restored by bone marrow grafting 4 weeks before infection. The level of serum interferon (IFN) of infected or uninfected W/W^v mice was comparable to that of +/+ mice. Therefore, impaired NK inducibility seems to be responsible, at least in part, for the high susceptibility of W/W^v mice to MCMV infection.     

Immunohistochemical localization of lactate dehydrogenase isoenzyme 1 in germ cell tumors of the testis

In this study, antiserum to lactate dehydrogenase isoenzyme 1 (LD 1) was used to determine immunohistochemical patterns of localization in a variety of germ cell neoplasms of the testis. All 29 seminomas and teratocarcinomas and four of seven embryonal carcinomas were positive for LD 1. These results suggest that elevated serum LD 1 levels noted in patients with germ cell neoplasms may be derived directly from neoplastic tissue and explain the relationship between serum LD 1 levels and tumor burden. A variety of neoplasms from other organs also were evaluated in order to determine the specificity of LD 1 staining for testicular tumors.     

Immunohistochemical localization and quantification of glucose transporters in the mouse brain

A family of seven facilitative glucose transporters (Glut1-5, 7 and 8) mediates the cellular uptake of glucose. In the brain, Glut2, Glut5 and Glut8 are found at relatively low levels whereas Glut1, Glut3 and Glut4 were reported in abundance in several brain regions. Using immunofluorescence, this study investigated, compared and quantified the localization of the brain major glucose transporters, Glut1, Glut3 and Glut4, in the different cerebral areas of CD1 mice. Most of the staining of Glut1, Glut3 and Glut4 in the mouse brain coincides with observations made in rats. The results confirm the cortical neuropil distribution of Glut3, the prominence of this transporter in the mossy fiber field of the hippocampus and the Glut3 and Glut4 immunostaining of the hippocampal pyramidal cell layer. The present study also reports novel localizations of the transporters such as the presence of Glut3 in neuronal perikarya, Glut4-labeled neurons in the CA3 of the hippocampus and the subiculum. In the cerebellum, Glut3 shows subcellular localization to the base of the Purkinje cell bodies near the axon hillock. Furthermore, an important population of Golgi cells was found to be strongly immunostained for Glut4 in the granular cell layer of the cerebellum. The quantification results suggest that the relative abundance of Glut1 in the frontal and motor cortices coincides well with the high-energy demands of these brain regions. However, the Glut4-selective abundance in cerebral motor areas supports its suggested role in providing the energy needed for the control of the motor activity. The reported neuropil distribution of Glut3 seems to uphold its suggested role in synaptic energy provision and neurotransmitter synthesis.We conclude that the cellular and regional distributions of the glucose transporters in the rodent brain seem to be relevant to their corresponding functions.     

Immunohistochemical localization of endothelin-like immunoreactivity in human tooth germ and mature dental pulp

Summary The distribution in oral tissues of endothelin, a multifunctional peptide originally identified within endothelial cells, and subsequently in some epithelial cells, neurons and neuroendocrine cells, has not been investigated yet. We have studied the localization of endothelin-like immunoreactivity in human tooth germ and mature dental pulp by immunohistochemical techniques. Such immunoreactivity was detected only within endothelial cells in both mature dental pulp and developing tooth. Arteries and veins of various sizes as well as small thin vessels displayed endothelin-like immunoreactivity. In the tooth germ, the cells of the enamel organ or the precursors of the odontoblasts were found unreactive. In the mature pulp, no cells of the stroma or nerves displayed endothelin-like immunoreactivity. These findings suggest that vascular endothelium may be the only source of endothelin in human dental tissues. It is tentatively proposed that endothelin released in mature tooth pulp may participate in the regulation of the pulpal blood flow. Although the possible role of endothelin in developing tissues is far from being clear, the mitogenic effects and the proto-oncogenes expression induced by endothelin in some cells raise the possibility that this peptide might also play a role during tooth development.     

Immunohistochemical localization of translationally controlled tumor protein in the mouse digestive system

Translationally controlled tumor protein (TCTP) is a housekeeping protein, highly conserved among various species. It plays a major role in cell differentiation, growth, proliferation, apoptosis and carcinogenesis. Studies reported so far on TCTP expression in different digestive organs have not led to any understanding of the role of TCTP in digestion, so we localized TCTP in organs of the mouse digestive system employing immunohistochemical techniques. Translationally controlled tumor protein was found expressed in all organs studied: tongue, salivary glands, esophagus, stomach, small and large intestines, liver and pancreas. The expression of TCTP was found to be predominant in epithelia and neurons of myenteric nerve ganglia; high in serous glands (parotid, submandibular, gastric, intestinal crypts, pancreatic acini) and in neurons of myenteric nerve ganglia, and moderate to low in epithelia. In epithelia, expression of TCTP varied depending on its type and location. In enteric neurons, TCTP was predominantly expressed in the processes. Translationally controlled tumor protein expression in the liver followed porto‐central gradient with higher expression in pericentral hepatocytes. In the pancreas, TCTP was expressed in both acini and islet cells. Our finding of nearly universal localization and expression of TCTP in mouse digestive organs points to the hitherto unrecognized functional importance of TCTP in the digestive system and suggests the need for further studies of the possible role of TCTP in the proliferation, secretion, absorption and neural regulation of the digestive process and its importance in the physiology and pathology of digestive process.     

Susceptibility of mast cell-deficient W/Wv mice to Cryptosporidium parvum.

Mast cell-deficient W/Wv infant mice were similar to normal mice in their susceptibility to and recovery from infection with the intestinal protozoan Cryptosporidium parvum. W/Wv adult mice were significantly more susceptible to primary infection than were normal adult mice, but both groups recovered at a similar rate.     

Immunohistochemical localization of murine stage-specific embryonic antigens in human testicular germ cell tumors.

Monoclonal antibodies raised against and/or recognizing stage-specific antigens on preimplantation mouse embryos and stem cells of murine teratocarcinoma were used to localize these antigens immunohistochemically on human testicular germ cell tumors. SSEA-1, the antigen found on mouse embryonal carcinoma (EC) cells and embryonic cells from the 8-cell stage embryo onward, including the fetal primordial germ cells, was detected on yolk sac carcinoma components of human tumors, but not on EC cells. SSEA-3, the antigen found on follicular ova, fertilized eggs, early cleavage stage embryonic cells, and visceral endodermal cells of the mouse embryo, but not on mouse EC cells, was detected on human EC cells. Both antigens were found on the cell surface of fetal testicular germ cells but not in the seminiferous tubules of adult human testes. These data point out differences between human and murine EC cells suggesting that human EC cells correspond developmentally to a less mature embryonic cell than the murine EC cells. The possible histogenesis of human germ cell tumors from primordial and/or fetal germ cells is briefly discussed.     

Immunohistochemical localization of ryanodine receptors in mouse central nervous system.

The distribution of ryanodine receptor-like immunoreactivity in the mouse central nervous system was studied using two antibodies raised against synthetic peptides. These peptides represented a region conserved between the cardiac and skeletal muscle forms and a region specific to the cardiac form. Western blotting analysis and [3H]ryanodine binding analysis showed ryanodine receptors are expressed in all the brain regions. The activity was prominent in hippocampus and cerebral cortex. Immunohistochemical study demonstrated that the ryanodine receptors were localized unevenly in somata. Some apical and proximal dendrites in some cells were also labeled. In hippocampus pyramidal neurons in CA2-3 region were more labeled than CA1 region. Immunohistochemical distribution revealed by two antibodies was essentially the same but the fibers were more immunoreactive with the antibody raised against the cardiac muscle ryanodine form. The localization of ryanodine receptors was quite different from that of inositol 1,4,5-trisphosphate receptors.