Introduction and objective: In Europe, nearly every sixth couple in the reproductive age is invol... more Introduction and objective: In Europe, nearly every sixth couple in the reproductive age is involuntarily childless. In about 30%, both male and female reveal fertility problems. In about 10% of infertile men, azoospermia is the underlying cause. As conventional therapeutic options are limited, surgical testicular sperm extraction (TESE) is necessary to obtain sperms for assisted reproductive techniques. Regarding the females, up to 30% of all idiopathic infertilities are due to alterations of the uterine tube So far, no imaging technique, which does not require any labelling, is available to evaluate the male and female genital tract at a microscopic level under in vivo conditions. Thus, the aim of this study was to investigate the potential of optical coherence tomography (OCT) as a non-invasive diagnostic tool in gynaecology and andrology. Material and Methods: Tissues samples from the bovine testis, epididymis, vas deferens, ovary, oviduct (ampulla and isthmus) and uterus were obtained immediately after slaughter (14 cows aged 3 to 8 years and 14 bulls aged 3 to 6 years; breeds: Holstein- Friesian, and Deutsches Fleckvieh). Imaging was done by using the US Food and Drug Administration (FDA) approved probe-based Niris Imaging System (Imalux, Cleveland, Ohio, USA) and the Telesto 1325 nm OCT System and Ganymede 930 nm OCT System (Thorlabs Inc., Dachau, Germany). All images obtained were compared to histological images after paraffin embedding and HE staining. Results: OCT imaging visualized the microarchitecture of the testis, epididymis, spermatic duct and the ovary, oviduct and uterus. Using the Thorlabs systems a axial resolution of approx. 5μm and lateral resolution of 8- 15μm could be achieved. Different optical tissue volumes could be visualized, which depends on the optical penetration depth of the wavelength of the system used. While the tissue volume observed by probe based Imalux-OCT is similar to the used Thorlabs systems, the optical resolution is reduced. By means of the microscopic OCT-system differentiation of testical tissue structures like content and diameter of seminiferous tubules and the epididymal duct was possible. Structures of the female oviduct, like the primary, secondary and tertiary folds including the typical epithelium consisting of secretory and ciliated cells were identified. Ampulla and isthmus were clearly differentiated by the height of the folds and the thickness of the smooth muscle layer. Imaging was successful both from the outside wall and from the inner lumen. After experience with microscopic OCT-structure identification such structures could also be identified by means of probe based OCT. Conclusions: Technical improvement of probe-based OCT up to a high-resolution level of nowadays-available OCT microscopic systems could open up new ways of in vivo imaging in the reproductive tract. Potential applications could be an OCT-guided testicular biopsy for improving sperm retrieval or microscopic evaluation of the oviduct by OCT-assisted fertiloscopy. The latter would provide a valuable tool to facilitate the decision of which type of assisted reproductive techniques might be preferred.
Sperm binding within the oviductal sperm reservoir plays an important role for reproductive succe... more Sperm binding within the oviductal sperm reservoir plays an important role for reproductive success by enabling sperm survival and maintaining fertilizing capacity. To date, numerous in vitro technologies have been established to measure sperm binding capacity to cultured oviductal cells or oviductal explants. However, these methods do not accurately represent the microenvironment and complex multi-molecular nature of the oviduct. In this paper, we describe a novel protocol for assessing sperm binding capacity in the tubal sperm reservoir using an ex vivo oviduct culture in the bovine model. This protocol includes the staining of frozen-thawed bovine spermatozoa with the DNA-binding dye Hoechst 33342, the co-incubation of stained sperm in closed segments of the oviduct and the visualization and quantification of bound spermatozoa by fluorescence microscopy. By generating overlays of multiple Z-stacks of randomly selected regions of interest (ROIs), spermatozoa bound in the sperm res...
On the occasion of the held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest spea... more On the occasion of the held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest speakers and audience) collectively felt the need to make a public statement on the general issue of male reproductive health. Our intention is to raise awareness of what we believe is a neglected area of research despite alarming situations around the world. The disclosure strategy desired by the co-authors is to bring it to the attention of the greatest number partly by considering co-publication in the various periodicals dealing with Reproductive Biology and Andrology. BaCA's editorial office accepted this mission and found it natural that our periodical, the official journal of the French Andrology Society (SALF), should carry this message.
Introducción La zona pelucida (ZP) es una matriz extracelular transparente, que rodea al ovocito ... more Introducción La zona pelucida (ZP) es una matriz extracelular transparente, que rodea al ovocito y al embrión mamífero en estadios tempranos de desarrollo. Constituye una capa homogénea que separa el oolema de la región más interna de las células foliculares, la corona radiata. Prolongaciones celulares de esas células atraviesan la ZP y forman, con la membrana plasmática del ovocito, los puentes celulares (gap juctions), ver capítulo desarrollo folicular en este libro. La ZP desempeña un rol de importancia en los diferentes pasos del proceso de la fecundación (Cuadro 1). Establece el contacto especie-específico de las células espermáticas, la unión de los espermatozoides capacitados y la inducción de la reacción acrosómica (Florman y Storey, 1982). En la mayoría de las especies la adhesión de los espermatozoides tiene lugar por medio de oligosacáridos de las glicoproteínas de la zona, que interaccionan con proteínas, que se unen a los glúcidos, complementarias de los espermatozoides...
Galactosyltransferase (GalTase) has been discovered on a variety of cells where it is believed to... more Galactosyltransferase (GalTase) has been discovered on a variety of cells where it is believed to be involved in cell-cell adhesion and cell-substratum adhesion as well as in metastasis of carcinoma cells. This immunohistochemical study was undertaken to identify the topography and the cellular distribution of GalTase in normal prostatic tissue, benign prostatic hyperplasia (11 cases), and prostatic carcinoma (26 cases). Immunoreactive GalTase was found to be exclusively associated with carcinoma cells and with premalignant epithelial cells in prostatic hyperplasia. In highly differentiated carcinomas, most of the carcinoma cells are positive for GalTase, whereas in poorly differentiated tumors, GalTase immunoreactivity was restricted to a subset of carcinoma cells with obviously invasive behavior. At the cellular level, GalTase was localized in the cytoplasm and at the cell membrane. In sections of normal prostatic tissue, as well as in unaltered acini of prostatic hyperplasic tiss...
The purpose of this study was to investigate the expression of two zona pellucida gene families Z... more The purpose of this study was to investigate the expression of two zona pellucida gene families ZPB (ZP3 alpha) and ZPC (ZP3 beta). Sections of ovaries from bovine fetuses, calves and cows were labelled with polyclonal antibodies. Immunopositive labelling was found in both the follicle cells and the oocyte. Labelling was dependent on the stage of development. The specific sequence of immunopositive reactions suggests that in the bovine fetus both the ovary and the follicle cells contribute to the production of the zona pellucida during pre- and postnatal development.
Introduction and objective: In Europe, nearly every sixth couple in the reproductive age is invol... more Introduction and objective: In Europe, nearly every sixth couple in the reproductive age is involuntarily childless. In about 30%, both male and female reveal fertility problems. In about 10% of infertile men, azoospermia is the underlying cause. As conventional therapeutic options are limited, surgical testicular sperm extraction (TESE) is necessary to obtain sperms for assisted reproductive techniques. Regarding the females, up to 30% of all idiopathic infertilities are due to alterations of the uterine tube So far, no imaging technique, which does not require any labelling, is available to evaluate the male and female genital tract at a microscopic level under in vivo conditions. Thus, the aim of this study was to investigate the potential of optical coherence tomography (OCT) as a non-invasive diagnostic tool in gynaecology and andrology. Material and Methods: Tissues samples from the bovine testis, epididymis, vas deferens, ovary, oviduct (ampulla and isthmus) and uterus were obtained immediately after slaughter (14 cows aged 3 to 8 years and 14 bulls aged 3 to 6 years; breeds: Holstein- Friesian, and Deutsches Fleckvieh). Imaging was done by using the US Food and Drug Administration (FDA) approved probe-based Niris Imaging System (Imalux, Cleveland, Ohio, USA) and the Telesto 1325 nm OCT System and Ganymede 930 nm OCT System (Thorlabs Inc., Dachau, Germany). All images obtained were compared to histological images after paraffin embedding and HE staining. Results: OCT imaging visualized the microarchitecture of the testis, epididymis, spermatic duct and the ovary, oviduct and uterus. Using the Thorlabs systems a axial resolution of approx. 5μm and lateral resolution of 8- 15μm could be achieved. Different optical tissue volumes could be visualized, which depends on the optical penetration depth of the wavelength of the system used. While the tissue volume observed by probe based Imalux-OCT is similar to the used Thorlabs systems, the optical resolution is reduced. By means of the microscopic OCT-system differentiation of testical tissue structures like content and diameter of seminiferous tubules and the epididymal duct was possible. Structures of the female oviduct, like the primary, secondary and tertiary folds including the typical epithelium consisting of secretory and ciliated cells were identified. Ampulla and isthmus were clearly differentiated by the height of the folds and the thickness of the smooth muscle layer. Imaging was successful both from the outside wall and from the inner lumen. After experience with microscopic OCT-structure identification such structures could also be identified by means of probe based OCT. Conclusions: Technical improvement of probe-based OCT up to a high-resolution level of nowadays-available OCT microscopic systems could open up new ways of in vivo imaging in the reproductive tract. Potential applications could be an OCT-guided testicular biopsy for improving sperm retrieval or microscopic evaluation of the oviduct by OCT-assisted fertiloscopy. The latter would provide a valuable tool to facilitate the decision of which type of assisted reproductive techniques might be preferred.
Sperm binding within the oviductal sperm reservoir plays an important role for reproductive succe... more Sperm binding within the oviductal sperm reservoir plays an important role for reproductive success by enabling sperm survival and maintaining fertilizing capacity. To date, numerous in vitro technologies have been established to measure sperm binding capacity to cultured oviductal cells or oviductal explants. However, these methods do not accurately represent the microenvironment and complex multi-molecular nature of the oviduct. In this paper, we describe a novel protocol for assessing sperm binding capacity in the tubal sperm reservoir using an ex vivo oviduct culture in the bovine model. This protocol includes the staining of frozen-thawed bovine spermatozoa with the DNA-binding dye Hoechst 33342, the co-incubation of stained sperm in closed segments of the oviduct and the visualization and quantification of bound spermatozoa by fluorescence microscopy. By generating overlays of multiple Z-stacks of randomly selected regions of interest (ROIs), spermatozoa bound in the sperm res...
On the occasion of the held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest spea... more On the occasion of the held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest speakers and audience) collectively felt the need to make a public statement on the general issue of male reproductive health. Our intention is to raise awareness of what we believe is a neglected area of research despite alarming situations around the world. The disclosure strategy desired by the co-authors is to bring it to the attention of the greatest number partly by considering co-publication in the various periodicals dealing with Reproductive Biology and Andrology. BaCA's editorial office accepted this mission and found it natural that our periodical, the official journal of the French Andrology Society (SALF), should carry this message.
Introducción La zona pelucida (ZP) es una matriz extracelular transparente, que rodea al ovocito ... more Introducción La zona pelucida (ZP) es una matriz extracelular transparente, que rodea al ovocito y al embrión mamífero en estadios tempranos de desarrollo. Constituye una capa homogénea que separa el oolema de la región más interna de las células foliculares, la corona radiata. Prolongaciones celulares de esas células atraviesan la ZP y forman, con la membrana plasmática del ovocito, los puentes celulares (gap juctions), ver capítulo desarrollo folicular en este libro. La ZP desempeña un rol de importancia en los diferentes pasos del proceso de la fecundación (Cuadro 1). Establece el contacto especie-específico de las células espermáticas, la unión de los espermatozoides capacitados y la inducción de la reacción acrosómica (Florman y Storey, 1982). En la mayoría de las especies la adhesión de los espermatozoides tiene lugar por medio de oligosacáridos de las glicoproteínas de la zona, que interaccionan con proteínas, que se unen a los glúcidos, complementarias de los espermatozoides...
Galactosyltransferase (GalTase) has been discovered on a variety of cells where it is believed to... more Galactosyltransferase (GalTase) has been discovered on a variety of cells where it is believed to be involved in cell-cell adhesion and cell-substratum adhesion as well as in metastasis of carcinoma cells. This immunohistochemical study was undertaken to identify the topography and the cellular distribution of GalTase in normal prostatic tissue, benign prostatic hyperplasia (11 cases), and prostatic carcinoma (26 cases). Immunoreactive GalTase was found to be exclusively associated with carcinoma cells and with premalignant epithelial cells in prostatic hyperplasia. In highly differentiated carcinomas, most of the carcinoma cells are positive for GalTase, whereas in poorly differentiated tumors, GalTase immunoreactivity was restricted to a subset of carcinoma cells with obviously invasive behavior. At the cellular level, GalTase was localized in the cytoplasm and at the cell membrane. In sections of normal prostatic tissue, as well as in unaltered acini of prostatic hyperplasic tiss...
The purpose of this study was to investigate the expression of two zona pellucida gene families Z... more The purpose of this study was to investigate the expression of two zona pellucida gene families ZPB (ZP3 alpha) and ZPC (ZP3 beta). Sections of ovaries from bovine fetuses, calves and cows were labelled with polyclonal antibodies. Immunopositive labelling was found in both the follicle cells and the oocyte. Labelling was dependent on the stage of development. The specific sequence of immunopositive reactions suggests that in the bovine fetus both the ovary and the follicle cells contribute to the production of the zona pellucida during pre- and postnatal development.
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