Neuropeptide Y1 receptors in the rat genital tract

Using in situ hybridization and immunohistochemistry, the expression of type 1 neuropeptide Y (NPY) receptors (Y1-Rs) has been demonstrated in the rat genital tract. In the male Y1-R mRNA and Y1-R-like immunoreactivity (LI) were found in smooth muscles of predominantly arterioles and small arteries inside testis. Fibers showing NPY-LI could not be detected within testis but only in the tunica albuginea. These Y1-Rs are suggested to mediate vasoconstriction, possibly activated by NPY released from nerves in the tunica albuginea. In the female rat Y1-R mRNA, but not Y1-R-LI was found in vascular smooth muscles of arteries in the ovary and oviduct. In the oviduct Y1-R mRNA was also detected in the non-vascular smooth muscle layer. Fibers showing NPY-LI were found around blood vessels both in the ovary and oviduct. In the female genital tract also Y1-Rs may thus be involved in regulatory mechanisms mediating, for example, vasoconstriction.     

Neuropeptide Y immunoreactivity in human cerebrospinal fluid

Neuropeptide Y (NPY) immunoreactivity has been determined in human cerebrospinal fluid (CSF). High pressure liquid chromatography revealed that the NPY immunoreactivity co-eluted with authentic NPY. The range of NPY levels was 108 +/- 18 pg/ml (mean +/- S.D.) in a group of 28 normal subjects. In five additional subjects NPY immunoreactivity was measured in 4 sequential 8 ml aliquots of CSF to determine whether a rostro-caudal gradient was present. No significant differences in NPY levels were detected between any of the 4 aliquots.     

Immunolocalization of angiotensin 1 converting enzyme in the human male genital tract by the avidin-biotin-complex method

Summary Immunoreactivity for Angiotensin 1 Converting Enzyme was investigated in a series of 12 fixed and paraffinembedded normal human genital tract specimens. The Avidin-Biotin-Complex immunoperoxidase method was used with overnight (12 h) incubation with a polyclonal antihuman kidney Angiotensin 1 Converting Enzyme antiserum. All tissues, including testis, different parts of epididymis, ductus deferens, prostate and seminal vesicles, demonstrated a staining pattern. Immunoreactivity was observed on the luminal surface of these epithelia especially on nonmotile stereocilia. An intracellular positivity was only observed in spermatids on the acrosomal cap. Besides, an immunologic identity of Angiotensin 1 Converting Enzyme located on the different epithelia of the human male genital tract, on the endothelial cells of vessels and on the proximal tubule brush border of the kidney was observed.     

Immunolocalization of angiotensin 1 converting enzyme in the human male genital tract by the avidin-biotin-complex method

Immunoreactivity for Angiotensin 1 Converting Enzyme was investigated in a series of 12 fixed and paraffin-embedded normal human genital tract specimens. The Avidin-Biotin-Complex immunoperoxidase method was used with overnight (12 h) incubation with a polyclonal antihuman kidney Angiotensin 1 Converting Enzyme antiserum. All tissues, including testis, different parts of epididymis, ductus deferens, prostate and seminal vesicles, demonstrated a staining pattern. Immunoreactivity was observed on the luminal surface of these epithelia especially on non-motile stereocilia. An intracellular positivity was only observed in spermatids on the acrosomal cap. Besides, an immunologic identity of Angiotensin 1 Converting Enzyme located on the different epithelia of the human male genital tract, on the endothelial cells of vessels and on the proximal tubule brush border of the kidney was observed.     

Hypochlorous acid-induced stress on human spermatozoa. A model for inflammation in the male genital tract

The fertilising ability of human spermatozoa may be impaired by inflammations of the genital tract, although details of these processes are still unknown. Hypochlorous acid (HOCl), an important product of myeloperoxidase released from stimulated neutrophils, induces a concentration-dependent increase in externalisation of phosphatidylserine in ejaculated human spermatozoa as revealed by fluorescence-activated cell sorting (FACS) analysis. The increase of annexin-V binding cells starts already at about 10(-5) mol/l HOCl, while a formation of lysophosphatidylcholines as detected by matrix-assisted laser desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is only found at HOCl concentrations higher than 10(-4) mol/l. Thus, changes in lipid composition of spermatozoa are unlikely responsible for the phosphatidylcholine (PS)-externalisation. These data gave concomitant evidence that HOCl itself leads to a dramatic damage of the cell membrane. Thus, the neutrophil-derived HOCl contributes to the deterioration of spermatozoa leading to diminished fertilisation ability.     

Immunohistochemical localization of spermadhesin AWN in the porcine male genital tract

Boar spermadhesin (AWN) is a 14-kDa multifunctional protein, attached to the surface of the spermatozoa and involved in sperm capacitation and zona pellucida binding. The cellular origin of AWN was previously unknown. Moreover, the region of the male genital tract in which AWN becomes attached to the surface of spermatozoa was also uncertain. By using monospecific polyclonal antibodies against AWN, the immunohistochemical distribution pattern of AWN epitopes has been investigated in tissue sections of the porcine male genital tract. Our study has revealed that AWN is synthesized in the rete testis and in the epithelium of the seminal vesicles. The latter are also the major contributors of seminal plasma AWN. In addition, immunoblotting analysis has shown that AWN is present on epididymal spermatozoa. Our results indicate that the cellular origin of spermadhesins is species-specific. The attachment of AWN to epididymal spermatozoa is probably important in developing the capacity for fertilization.     

Immunocytochemical localization of uteroglobin in the genital tract of male rabbits

Summary Using the immunoperoxidase technique and specific goat antiserum to uteroglobin, selective immunochemical staining products are localized in the epididymal epithelium of the caput and proximal corpus region, at the adluminal border of the cauda epididymidis and, as well known, in epithelial cells of the endometrium of pregnant and progesterone-treated rabbits. Specific staining is also seen on spermatozoa. A uteroglobin-like antigen has been similarly localized in alveolar and bronchial epithelial cells of the lung. Testis, prostate, seminal vesicle and ductus deferens do not seem to contain in their tissues immunoreactive uteroglobin-like antigens. Similarly, the uterus and ductus epididymidis of immature rabbits are devoid of immunoreactivity. The presence of uteroglobin-like antigens in tissues other than the endometrium, particularly the ductus epididymidis, stimulates new discussions on the function of this protein in reproductive physiology and fertility research.     

Distribution of acid glycosidases in the male genital tract of the pheasant

The distribution of the activity of seven acid glycosidases in the reproductive organs of the pheasant (Phasianus colchicus) was studied. The study was carried out on seven mature birds at the age of 11 months during the reproductive season (May). Significant (p<0.01) differences in acid glycosidase activity dependent on enzyme origin were observed. Generally, the highest activity of acid glycosidases was found in the epididymis, intermediate in the ductus deferens and the lowest in the testes. Exceptionally, alpha-mannosidase had the highest activity in the ductus deferens. Anion-exchange chromatography elution profiles of most enzymes from the tested reproductive organs were similar, however evident differences were observed for beta-mannosidase forms.     

Amylase in human lungs and the female genital tract

Summary We investigated the localization of amylase in normal human lungs and the female genital tract using immunohistochemical and histochemical methods. Immunohistochemical procedures were applied to formaldehydefixed, paraffin-embedded specimens as well as to cryostat sections of periodate/lysine/paraformaldehyde (PLP)-fixed tissues. The starch-substrate-film method was used for the histochemical investigation of unfixed frozen sections. Amylase immunoreactivity was observed in ciliated epithelial cells of the bronchus and in serous cells of the bronchial glands but not in the alveolar epithelium. Immunoreactive amylase was also found in the cytoplasm of the ciliated epithelium of the fallopian tubes, especially in the apical part of the cytoplasm and in ciliary vesicles. Immunoreactive amylase was also found to be present in the surface epithelial cells and glands of the uterine cervix, as well as in the superficial part of the endometrial glands. The distribution of amylase activity revealed using histochemistry was similar to that observed in cryostat sections of PLP-fixed tissues after immunohistochemical staining. Amylase amtigenicity was better preserved in cryostat sections of PEP-fixed materials than in formaldehyde-fixed, paraffinembedded specimens. The results are discussed in relation to pulmonary and female-genital-tract diseases.This work was supported by a grant from the Tokyo Metropolitan Government     

Neuropeptide Y Y2 receptor signalling mechanisms in the human glioblastoma cell line LN319

Neuropeptide Y (NPY) regulates neurotransmitter release through activation of the Y2 receptor subtype. We have recently characterized a human glioblastoma cell line, LN319, that expresses exclusively NPY Y2 receptors and have demonstrated that NPY triggers transient decreases in cAMP and increases in intracellular calcium responses. The present study was designed to further characterize calcium signalling by NPY and bradykinin (BK) in LN319 cells. Both agonists elevated free intracellular calcium ([Ca(2+)](i)) without soliciting calcium influx. NPY appeared to activate two distinct signalling cascades that liberate calcium from thapsigargin- and ryanodine-insensitive compartments. One pathway proceeded through phospholipase C (PLC)-dependent phosphatidylinositol turnover, while the other triggered calcium release through a so far unidentified mediator. Part of the response was sensitive to pertussis toxin (PTX) under conditions where the toxin totally abolished the NPY-mediated effects on cAMP. The calcium release induced by BK on the other hand was largely PTX-insensitive, PLC-dependent, and from both thapsigargin- and ryanodine-sensitive stores. Following stimulation with NPY, subsequent [Ca(2+)](i) responses to NPY were strongly depressed. Partial heterologous desensitization occurred, when BK was used as the first agonist, whereas NPY had no effect on a subsequent stimulation with BK. These data suggest that NPY-induced calcium mobilization in LN319 cells involves two different G proteins and signalling mediators, and a hitherto unidentified calcium compartment. Homologous desensitization of NPY signalling might be explained by receptor-G protein uncoupling, while heterologous desensitization by BK could be the result of either transient depletion or inhibition of a mediator in the calcium signalling cascades activated by NPY.     

Pre-natal innervation of the human female genital tract

In the human fetus of 14 weeks, ganglia on either sides of the Müllerian uterovaginal canal contained two types of cells. In the 16th week, axons invaded the basal zone of the stratified squamous epithelium at the sides of the upper vagina. In the 20th week, vesicular nuclei typified the large neurons in the midportion of the cervico-vaginal ganglion. During the 22nd week, capsulated ganglia invaded the wall of the upper vagina forming three concentrically disposed strata. Non-capsulated clusters invaded its lamina propria. At the 24th week, axons were shaded after reaching the superficial zone of the stratified vaginal epithelium. In the 28th week, satellites surrounded the mature neurons and sheath cells enveloped the axons. Ganglia invaded the splitted muscle layer of the upper vagina at 30 weeks. Intraepithelial fibres invaded the whole thickness of the endometrium, the columnar epithelium of the cervix and uterine tube at 40 weeks. Nerve cells were detected among the basal epithelial cells of the lower vagina and its subepithelial plexus.     

Neuropeptide Y is a vasoconstrictor in human nasal mucosa.

Neuropeptide Y (NPY) is a neurotransmitter in sympathetic nerve fibers in human nasal mucosa. Like norepinephrine, NPY acts as a vasoconstrictor. An established method of nasal provocation was used to determine the effects of topically applied NPY on nasal resistance to airflow measured by anterior rhinomanometry, the protein content of nasal secretions, and the protein content of bradykinin-induced secretions. NPY (2.3 nmol) reduced the resistance to inspiratory airflow by 57 +/- 18% (P < 0.001) in 10 normal subjects and by 50 +/- 17% (P < 0.05) in 12 subjects with perennial rhinitis. In nasal provocations, NPY in doses of 0.1-10 nmol had no effect on vascular (albumin), glandular (lysozyme, glycoconjugate), or total proteins present in lavaged nasal secretions. Because the vasoconstrictor properties of NPY may only be apparent in the presence of increased vascular permeability and albumin exudation, bradykinin (BK) nasal provocation was performed. BK (500 nmol) significantly increase total protein (10- to 20-fold), albumin (10- to 30-fold), and glycoconjugate (2- to 5-fold) in lavage fluid. NPY (2.3 nmol) reduced BK-induced total protein by 59 +/- 15% (P < 0.05) and albumin by 63 +/- 17% (P < 0.02) but had no significant effect on glandular secretion. Therefore exogenous administration of NPY to the human nasal mucosa reduced nasal airflow resistance and albumin exudation without affecting submucosal gland secretion. NPY agonists may be useful for the treatment of mucosal diseases characterized by vasodilation, vascular permeability, and plasma exudation.     

On the Y fluorescence in human male fibroblasts

Summary In a system of mitotically synchronized human fibroblast cells of the karyotype 46, XY and 47,XYY, the frequency and the variation in structure of the quinacrine dihydrochloride stained heterochromatic part of the Y chromosome was counted stepwise in regard to the periods of the cell cycle. There was no change of frequency and structure in correlation to these periods, especially no visible uncoiling in relation to the time of DNA synthesis.

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Zusammenfassung In einem System teilungssynchroner Kulturen menschlicher Fibroblasten des Karyotyp 46,XY und 47,XYY wurden Häufigkeiten und Strukturen des mit Quinacrindihydrochlorid gefärbten heterochromatischen Anteils des Y-Chromsoms in Abhängigkeit von den einzelnen Phasen des Zellcyclus fortlaufend beobachtet. Es fand sich keine Veränderung in Korrelation zu den Cyclusphasen, insbesondere keine meßbare Entspiralisation des heteropyknotischen F-body in Verbindung mit dem Zeitraum des DNS-Synthese.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.