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Distribution of surface anionic sites during formation of mouse hybrid myelomas
Abstractsofl6th AnnualScientificMeetingof
448
the seminal vesicle. 2. The adult cell or spermiophore,which is found in the female soon after copulation. 3. The mature sperm after capacitation on its way to the ovary. The prospermium has the form of a tube, turned up like a cuff over its whole length. Its outermost shell is a trilaminar unit membrane to which the nucleus is attached near the anterior end of the prospermium. The length of the spermiophore is about twice that of the prospermium since at this stage the tube is unfolded to its full length. Together with the unfolding of the tube, the outermost membrane invaginates into the posterior end of the sperm cell to form the acrosomal canal. During this invagination, the nucleus is carried into the posterior part of the spermiophore. Finally, during capacitation the acrosomal canal evaginates from the spermiophore, carrying with it the nucleus, which is thus exposed to the exterior.
THE GLYCOCALYX OF THE RAM SPERM CELL: DETERMINATION OF CATIONIZED FERRITIN HYDRAZIDE AND CONCANAVALIN A-FERRITIN BINDING SITES B. Bartoov, Department University,
F. Eltes
and 2. Malik
of Life Sciences, Ramat Gan, Israel
Bar-Ilan
Surface glycoproteins of the sperm cell reflect the epididymal maturation state and may play a major role in motility and effective fertilization. The purpose of the present study was to localize negatively charged glycoproteins, sialic acid, glucose and mannose residues of the ram sperm glycocalyx. Cationized ferritin labelled the negatively charged residues of the outer membrane of the entire sperm cell: head, The binding site was very neck and tail. closely related to the membrane and seen prominently in the mitochondrial region. Ferritin adipic acid dihydrazide (FHZ) labelled mostly sialic acid residues of the post-acrosomal region, though labelled patches could be detected on No binding was found in the acrosome. FHZ particles the mitochondrial region. were closely attached to the postacrosomal membrane, whereas the patches on the acrosomal membrane were 40 nm Poisoning the cell with cacodylapart. ate buffer exposed all parts of the cell The to FHZ binding including the tail. poisoned cell was extraordinarily labelled even at the intra-mitochondrial Con Aspaces and nuclear envelope.
TheIsraelSociely
ferritin was localized in a multi-layer order on the cell membrane including the tail. Glucose-bearing glycoproteins appeared as different macromolecules ranging in length from 10 to 60 nm.
DISTRIBUTION OF SURFACE ANIONIC SITES DURING FORMATION OF MOUSE HYBRID MYELOMAS Y. Marikovsky,
L. Wang*
and M. Inbar*
Department of Membrane Research, Weizmann Institute of Science; and *Department of Cell Biology, MilesYeda Ltd., Rehovot, Israel
The advent of the hybrid-myeloma technique for the production of monoclonal antibodies has provided a new model system to study membrane properties as each hybridoma results from the fusion of a normal spleen cell with a malignant myeloma cell. By employing the cationized ferritin (CF) labelling method, we studied the surface anionic site distribution on membranes of mouse normal spleen cells obtained from BALB/c mice after immunization with L-thyroxine (BSA-T4) and cells from a mouse myeloma line (NS-1) --the two parental cells of an anti-BSA-T4 antibody producing The aim of hybridoma cell line (16A). the study was to determine, using surface topography, whether the surface charge characteristic of the newly created hybridoma cell line is determined by its parental cells. Pre-fixation of cells with glutaraldehyde immobilizes CF-binding sites and thus prevents their relocation and regrouping. Nevertheless, the NS-1 myeloma cells exhibit patchA similar pattern of like heaps of CF. patch-like CF heaps is exhibited on the The main dishybridoma cell line 16A. tinction between these cells and that of a normal spleen cell is that the latter do not show any heaps of CF on even and continuous CF-labelled surfaces. The distribution of CF specific sites into heaps on the 16A hybridoma cell membrane may indicate that the surface anionic properties of the malignant myeloma cell become dominant in the cell hybrid.
SOME ULTRASTRUCTURAL ASPECTS OF REGENERATION FOLLOWING INJURY OF RAT GASTROCNEMIUS MUSCLE U. Oron
and D. Roth
Department Faculty of University,
of Zoology, George S. Wise Life Sciences, Tel dviv Tel Aviv 69978, Israel
448
the seminal vesicle. 2. The adult cell or spermiophore,which is found in the female soon after copulation. 3. The mature sperm after capacitation on its way to the ovary. The prospermium has the form of a tube, turned up like a cuff over its whole length. Its outermost shell is a trilaminar unit membrane to which the nucleus is attached near the anterior end of the prospermium. The length of the spermiophore is about twice that of the prospermium since at this stage the tube is unfolded to its full length. Together with the unfolding of the tube, the outermost membrane invaginates into the posterior end of the sperm cell to form the acrosomal canal. During this invagination, the nucleus is carried into the posterior part of the spermiophore. Finally, during capacitation the acrosomal canal evaginates from the spermiophore, carrying with it the nucleus, which is thus exposed to the exterior.
THE GLYCOCALYX OF THE RAM SPERM CELL: DETERMINATION OF CATIONIZED FERRITIN HYDRAZIDE AND CONCANAVALIN A-FERRITIN BINDING SITES B. Bartoov, Department University,
F. Eltes
and 2. Malik
of Life Sciences, Ramat Gan, Israel
Bar-Ilan
Surface glycoproteins of the sperm cell reflect the epididymal maturation state and may play a major role in motility and effective fertilization. The purpose of the present study was to localize negatively charged glycoproteins, sialic acid, glucose and mannose residues of the ram sperm glycocalyx. Cationized ferritin labelled the negatively charged residues of the outer membrane of the entire sperm cell: head, The binding site was very neck and tail. closely related to the membrane and seen prominently in the mitochondrial region. Ferritin adipic acid dihydrazide (FHZ) labelled mostly sialic acid residues of the post-acrosomal region, though labelled patches could be detected on No binding was found in the acrosome. FHZ particles the mitochondrial region. were closely attached to the postacrosomal membrane, whereas the patches on the acrosomal membrane were 40 nm Poisoning the cell with cacodylapart. ate buffer exposed all parts of the cell The to FHZ binding including the tail. poisoned cell was extraordinarily labelled even at the intra-mitochondrial Con Aspaces and nuclear envelope.
TheIsraelSociely
ferritin was localized in a multi-layer order on the cell membrane including the tail. Glucose-bearing glycoproteins appeared as different macromolecules ranging in length from 10 to 60 nm.
DISTRIBUTION OF SURFACE ANIONIC SITES DURING FORMATION OF MOUSE HYBRID MYELOMAS Y. Marikovsky,
L. Wang*
and M. Inbar*
Department of Membrane Research, Weizmann Institute of Science; and *Department of Cell Biology, MilesYeda Ltd., Rehovot, Israel
The advent of the hybrid-myeloma technique for the production of monoclonal antibodies has provided a new model system to study membrane properties as each hybridoma results from the fusion of a normal spleen cell with a malignant myeloma cell. By employing the cationized ferritin (CF) labelling method, we studied the surface anionic site distribution on membranes of mouse normal spleen cells obtained from BALB/c mice after immunization with L-thyroxine (BSA-T4) and cells from a mouse myeloma line (NS-1) --the two parental cells of an anti-BSA-T4 antibody producing The aim of hybridoma cell line (16A). the study was to determine, using surface topography, whether the surface charge characteristic of the newly created hybridoma cell line is determined by its parental cells. Pre-fixation of cells with glutaraldehyde immobilizes CF-binding sites and thus prevents their relocation and regrouping. Nevertheless, the NS-1 myeloma cells exhibit patchA similar pattern of like heaps of CF. patch-like CF heaps is exhibited on the The main dishybridoma cell line 16A. tinction between these cells and that of a normal spleen cell is that the latter do not show any heaps of CF on even and continuous CF-labelled surfaces. The distribution of CF specific sites into heaps on the 16A hybridoma cell membrane may indicate that the surface anionic properties of the malignant myeloma cell become dominant in the cell hybrid.
SOME ULTRASTRUCTURAL ASPECTS OF REGENERATION FOLLOWING INJURY OF RAT GASTROCNEMIUS MUSCLE U. Oron
and D. Roth
Department Faculty of University,
of Zoology, George S. Wise Life Sciences, Tel dviv Tel Aviv 69978, Israel