Distribution Of Intramembranous Particles Research Articles

Post-testicular changes in the density and distribution of intramembrane particles of stallion sperm surface domains

Freeze-fracture replicas of stallion spermatozoa, collected from the proximal caput, corpus and cauda epididymides regions, were analyzed by electron microscopy to explore the distribution and density of intramembrane particles (IMP). Conspicuous differences in density and arrangement of the IMP were observed in the different topographical domains of mature and immature spermatozoa. A reduction of IMP, especially remarkable in the post-acrosomal domain, was observed in mature epididymal spermatozoa when compared with samples collected from ductuli efferentes. Some structural species-specific differences were also observed. The significance of these changes has not been determined, but remodeling of membrane components during developmental processes constitutes a fine control mechanism to ensure that key molecules are in the correct membrane position and during an appropriate timeframe to mediate fertilization.

Specificity of membrane specializations in mechanoreceptors of birds—A freeze-etching study

The detailed knowledge of the molecular process of mechanotransduction is still an unsolved question. The investigation of the intramembranous structure of the cutaneous mechanoreceptors may play an important role in elucidating this problem. In this relation, Herbst sensory corpuscles in ducks were studied for the first time using the freeze-etching and thin sectioning techniques. Herbst corpuscles have the basic structural components valid for most of the encapsulated mechanoreceptors in mammals: a capsule made of perineural cells, a lamellar complex of modified Schwann cells, surrounding the non-myelinated part of the receptor nerve fiber and its ending. Freeze-etching replicas reveal that the plasmalemmae of the capsule cells, modified Schwann cells and axolemmae of parts of the nerve fiber differ in both density and pattern of distribution of intramembranous particles (IMPs) as well as IMP size. On all the plasmalemmae the IMP density is higher on the P-face (2000–3300 µm−2) than the respective E-face (800–1500 µm−2). The axolemma of the ending of the receptor nerve fiber expresses higher density of IMPs than its shaft. The mean IMP size for all the plasmalemmae varies between 5.5 and 7.5 nm. Many tight junctions occur between the capsule cells. These results indicate that the non-myelinated axolemma as well as the plasmalemmae of other components of Herbst corpuscles are specialized in terms of content and distribution of IMPs. The IMPs may represent various kinds of mechanosensitive channel proteins or related membrane-bound proteins participating in the process of mechanotransduction.

Differences in intramembrane particle distribution in young and old human erythrocytes.

The distribution of intramembrane particles in human erythrocytes was studied by freeze-fracture on young and old cells and compared to that obtained after ATP depletion or following addition of a clustering agent. It was shown that intramembrane particles became aggregated and the mean particle density increased as the cells aged. Likewise, both particle aggregation and increased density were found in young cells after moderate ATP depletion. In contrast, mean particle density was markedly reduced in both cell types after exhaustive depletion. Paradoxically, Zn treatment led to decreased particle density in young cells, whilst producing the opposite effect in aged cells. The results suggest that their low ATP content may account for the increased particle density of senescent cells.

Cold-induced imbibition damage of lettuce embryos: a study using cryo-scanning electron microscopy

The impact of rehydration on a multicellular organism was studied in lettuce (Lactuca sativaL.) embryos, using cryo-scanning electron microscopy (cryo-SEM). Naked embryos were sensitive to imbibitional stress, whereas embryos with an intact, thick-walled endosperm were not. Imbibitional injury to naked embryos was mainly confined to the outer 2–3 cell layers of the axis. These cells failed to swell and appeared poorly hydrated. Deeper layers were not affected even after extended periods of cold rehydration. The proportion of damaged cells (6–7% of total) roughly corresponded with the additional K+that gradually leached from the embryos. Damaged embryos were able to survive the loss of their surface layers and form adventitious roots. The swelling of inner tissues caused the dead surface layers to rupture into patches. Plasma membranes in dried embryos showed normal bilayer structure with a homogeneous distribution of intra-membrane particles (IMPs), also after non-injurious rehydration. Imbibitionally damaged plasma membranes showed many irregularities, such as globular insertions, that probably resulted from malfusions in the ruptured membrane, but the IMPs were still randomly distributed.

Ultrastructural analysis of differentiation in Legionella pneumophila.

Legionella pneumophila is an adaptive pathogen that replicates in the intracellular environment of fundamentally divergent hosts (freshwater protozoa and mammalian cells) and is capable of surviving long periods of starvation in water when between hosts. Physiological adaptation to these quite diverse environments seems to be accompanied by morphological changes (Garduño et al., p. 82-85, in Marre et al., ed., Legionella, 2001) and conceivably involves developmental differentiation. In following the fine-structural pathway of L. pneumophila through both in vitro and in vivo growth cycles, we have now discovered that this bacterium displays an unprecedented number of morphological forms, as revealed in ultrathin sections and freeze-fracture replicas for transmission electron microscopy. Many of the forms were identified by the obvious ultrastructural properties of their cell envelope, which included changes in the relative opaqueness of membrane leaflets, vesiculation, and/or profuse invagination of the inner membrane. These changes were best documented with image analysis software to obtain intensity tracings of the envelope in cross sections. Also prominent were changes in the distribution of intramembranous particles (clearly revealed in replicas of freeze-fractured specimens) and the formation of cytoplasmic inclusions. Our results confirm that L. pneumophila is a highly pleomorphic bacterium and clarify some early observations suggesting sporogenic differentiation in L. pneumophila. Since morphological changes occurred in a conserved sequence within the growth cycle, our results also provide strong evidence for the existence of a developmental cycle in L. pneumophila that is likely accompanied by profound physiological alterations and stage-specific patterns of gene expression.

Ultrastructural characterization of theisolated hydrogenosome in Tritrichomonas foetus

In the present study we show new aspects of the hydrogenosome ultrastructure as wellalterations induced by the fractionation technique. The morphology of freshly isolated hydrogenosomes as well those found in whole cells of Tritrichomonas foetus were examined in thin-sections, in replicas of fast-freezing, and conventional freeze-fracture, freeze-etching, and by high resolution scanning electron microscopy (field emission in-lens scanning electron microscopy). The true surface as well the concave and convex fracture faces of the inner and outer membranes are shown. We showed that after fractionation procedures the hydrogenosome ultrastructure can be changed, since isolated hydrogenosomes present patchwork-like structures, rosettes and the inner hydrogenosomal membrane is displaced. The peripheral vesicle is seen as a distinct compartment, since its content and morphological appearance is quite different from the rest of the organelle. The peripheral vesicle shows a smooth surface but presenting pores with 20 nm in diameter with a density of 7/μm 2 when observed after freeze-etching. We report the existence of characteristic intramembrane particles distribution and density on hydrogenosome membranes of isolated and whole T. foetus, suggesting that this organelle can have its morphology changed as consequence of technical modifications or as expression of its metabolic state.

Quantal acetylcholine release: vesicle fusion or intramembrane particles?

Images of vesicle openings in the presynaptic membrane have regularly been shown to increase in number after stimulation of cholinergic nerves. However, with a very few exceptions, the occurrence of vesicle openings is delayed in time with respect to the precise moment of transmitter release. In contrast, a transient change in the size and distribution of intramembrane particles (IMPs) has constantly been found as a characteristic change affecting the presynaptic membrane in a strict time coincidence with the release of acetylcholine quanta. This is illustrated here in a rapid-freezing experiment performed on small specimens of the Torpedo electric organ during transmission of a single nerve impulse. A marked change affected IMPs in the presynaptic membrane for 3-4 ms, i.e., a population of IMPs larger than 10 nm momentarily occurred in coincidence with the passage of the impulse. The nicotinic receptors, abundantly visible in the postsynaptic membranes, also underwent very fleeting structural changes during synaptic transmission. In conclusion, for rapidly operating neurotransmitters like acetylcholine, a characteristic IMP change was regularly found to coincide in the presynaptic membrane with the production of neurotransmitter quanta, whereas images of vesicles fusion were either delayed or even dissociated from the release process. This is discussed in connection to the different modes of release recently described for other secreting systems.

Synaptic architecture of glomeruli in lamina II of the chicken spinal cord, as revealed using ultrathin section and freeze fracture techniques.

Synaptic glomeruli in lamina II of the chicken dorsal horn were studied using the freeze fracture technique, and the results were compared with those obtained using the ultrathin section technique. Our findings using the freeze fracture technique were as follows. (1) On the presynaptic P-face of the central terminal, intramembrane particles (IMPs) were arranged circularly around a small dimple which was reported to be a synaptic vesicle attachment site. A distinct area with aggregated large IMPs was found on the postsynaptic E-face of some peripheral neuronal elements. (2) The area with small IMPs intermingled with several dimples and the area with aggregated large IMPs were present juxtaposed on the same central terminal P-face. The area with aggregated large IMPs indicates that the central terminal functions as a postsynaptic element; accordingly, the two areas represent a reciprocal synapse. (3) Distinct IMP aggregates were observed on the P-face of vesicle-containing dendrites which did not face the central terminal. (4) A fractured septate junction was revealed as numerous parallel-lined furrows on the E-face of the central terminal. The distribution of IMPs in the synaptic glomerulus supports the hypothesis that the synaptic glomerulus is the site of the local inhibitory feedback circuit for pain transmission.

Membrane Particle Distribution in the Sternal Epithelia of the Terrestrial Isopod Porcellio scaber Latr. (Crustacea, Oniscidea) during CaCO3 Deposit Formation and Resorption, a Freeze-Etch Analysis

The anterior sternal epithelium of terrestrial isopods transports cuticular Ca2+ to and from large sternal CaCO3 deposits. We analyzed the anterior and posterior sternal epithelium by the means of the freeze-etch technique and measured the size distribution and density of intramembrane particles (IMPs) during three different molting stages. At least three IMP size classes around 4.5, 7.7, and 9.4 nm can be distinguished on the P-face of the apical and basolateral plasma membrane. An additional size class of around 12.8 nm is restricted to the apical compartment. In the anterior sternal epithelium, the density of these large particles changes by a factor of 1.9 during the molt cycle, suggesting a role in CaCO3 formation and/or resorption. The density of the smaller IMPs rises transiently by a factor of 1.3 in the posterior sternal epithelium only. The IMP density of the basolateral plasma membrane increases significantly by a factor of 1.4 and 1.3 in the anterior and posterior sternal epithelia, respectively. The results indicate that increases in the IMP density contribute to the differentiation to an increased transport activity during the cyclic enlargements of the plasma membrane surface area in the anterior sternal epithelium.

Freeze-fracture and cytochemical evidence for structural and functional alteration in the axolemma and myelin sheath of adult guinea pig optic nerve fibers after stretch injury.

Recent work in animal models of human diffuse axonal injury has generated the hypothesis that, rather than there being physical disruption of the axolemma at the time of injury, a pertubation of the membrane occurs, which leads, over time, to a dysfunction of the physiology of the axolemmal. This dysfunction is posited to lead to a disruption of ionic homeostasis within the injured axon, leading to secondary axotomy some hours after the initial insult. We decided to test the hypothesis that membrane pump/ion channel activity or function is compromised and this would be reflected in structural changes within the axolemma and myelin sheath. We used freeze fracture and cytochemical techniques to provide evidence for change in membrane structure and the activity of membrane pumps after nondisruptive axonal injury in the adult guinea pig optic nerve. Within 10 min of injury, structural changes occurred in the distribution and number of intramembranous particles (IMPs) in the internodal axolemma. By 4 h, there was novel labeling for Ca-ATPase membrane pump activity at the same site. There was loss of IMPs from the nodal axolemma extending over several hours after injury. There was loss of both membrane pump Ca-ATPase and p-nitro-phenylphosphatase (p-NPPase) activity of the node. There was loss of ecto-Ca-ATPase activity but increased labeling for p-NPPase activity at sites of dissociation of compacted myelin. Quantitative freeze-fracture demonstrated statistically significant changes in membrane structure. We provide support for the hypothesis that structural and functional changes occur in the axolemma and myelin sheath at nondisruptive axonal injury.

Membrane structure of caveolae and isolated caveolin-rich vesicles.

Caveolae are specialized invaginated domains of the plasma membrane. Using freeze-fracture electron microscopy, the shape of caveolae and the distribution of intramembrane particles (integral membrane proteins) were analyzed. The caveolar membrane is highly curved and forms flask-like invaginations with a diameter of 80-120 nm with an open porus of 30-50 nm in diameter. The fracture faces of caveolar membranes are nearly free of intramembrane particles. Protein particles in a circular arrangement surrounding the caveolar opening were found on plasma membrane fracture faces. For isolation of caveolin-enriched membrane vesicles, the method of Triton X-100 solubilization, as well as a detergent-free isolation method, was used. The caveolin-rich vesicles had an average size of between 100 and 200 nm. No striated coat could be detected on the surface of isolated caveolin-rich vesicles. Areas of clustered intramembrane particles were found frequently on membrane fracture faces of caveolin-rich vesicles. The shape of these membrane protein clusters is often ring-like with a diameter of 30-50 nm. Membrane openings were found to be present in the caveolin-rich membrane vesicles, mostly localized in the areas of the clustered membrane proteins. Immunogold labeling of caveolin showed that the protein is a component within the membrane protein clusters and is not randomly distributed on the membrane of caveolin-rich vesicles.

Sequence-related behaviour of transmembrane domains from class I receptor tyrosine kinases

2 H NMR spectroscopy and freeze-fracture electron microscopy were used to compare the transmembrane domains of two Class I protein receptor tyrosine kinases (the EGF receptor and Neu/erbB-2) regarding overall behaviour in fluid lipid bilayer membranes. The 34-residue peptide, EGFR tm, was synthesised to contain the 23 amino acid hydrophobic stretch (Ile 622 to Met 644) thought to span the membrane of the human EGF receptor, plus the first 10 amino acids (Arg 645 to Thr 654) of the cytoplasmic domain. Deuterium probes replaced selected 1 H nuclei at sites corresponding to Ala 623, Met 644, and Val 650. The 38-residue peptide, Neu tm, was synthesised having the 21 residue hydrophobic stretch (Ile 660 to Ile 680) calculated to span the membrane in rat Neu/erbB-2, plus residues Lys 681 to Thr 691 of the contiguous cytoplasmic domain. Deuterium probes replaced selected 1 H nuclei at Ala 661, Leu 667, and Val 676. A third peptide, Neu tm*, was also prepared, corresponding to the transmembrane domain of a constitutively-activating Neu/erbB-2 transformant in which Val 664 is replaced by Glu: it was deuterated in a manner identical to Neu tm. Peptides were studied by 2 H NMR spectroscopy at 1 mol% and 6 mol% in unsonicated fluid bilayers of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) and in POPC containing 33 mol% cholesterol, over the range 12° to 65°C. Overall motion was found to be different for each of the three peptides under a given set of conditions. EGFR tm spectra were characteristic of axially symmetric motion in membranes of POPC alone, and in POPC/cholesterol at 35°C and above. In contrast, spectra of the transmembrane peptides, Neu tm and Neu tm*, were characteristic of significantly axially asymmetric motion under all conditions studied (and regardless of sample preparation method). Addition of 33% cholesterol to membranes was accompanied by spectral changes consistent with increased formation of peptide dimers/oligomers in all cases. The transformant peptide, Neu tm*, showed greater spectral evidence of immobilisation than did the wild type - probably reflecting a greater tendency to form large oligomers. Sequence-related details within the transmembrane domains of Class I receptor tyrosine kinases appear to exert important control over their associations within membranes. Freeze-fracture electron microscopy of the NMR samples demonstrated their liposomal nature. Peptide-related intramembranous particles (IMPs) were present which likely represent oligomers of the transmembrane peptide. IMP size and distribution were similar under a given set of conditions for all three peptides, suggesting that the differences seen by NMR spectroscopy reflect structures smaller than the 2 nm resolution limit of freeze-fracture EM and peptide relationships within its 20 nm accuracy of identifying lateral position.

A freeze-fracture study of the membrane morphology of phosphatidylethanolamine-deficient Escherichia coli cells

Freeze-fracture electron microscopy was applied to study membrane morphology in a phosphatidylethanolamine-deficient E. coli strain. For growth, this strain requires millimolar concentrations of specific divalent cations like Mg 2+ or Ca 2+. These cations bring the bilayer to nonbilayer phase transition temperature of the lipids back to wild type levels by shifting the phase preference of cardiolipin in the membrane towards the inverted hexagonal (H II) phase. Under growth conditions, these cells show a bilayer based membrane with an intramembrane particle distribution as in wild type cells. Upon lowering the temperature, smooth areas are observed corresponding to gel state lipid bilayer domains. Ca 2+ was used to manipulate the phase behavior of the membrane lipids in situ. Exposing the cells to Ca 2+ up to 100 mM at 42°C did not result in the appearance of nonbilayer structures, despite the fact that in total lipid extracts under these conditions the hexagonal H II phase was observed. However, the addition of a Ca 2+ ionophore, which leads to exposure to Ca 2+ of both faces of the plasma membrane, gives rise to formation of H II phase, stacked bilayer domains and blebbing upon addition of 50 mM CaCl 2 at 42°C. We conclude that the asymmetrical localization of divalent cations in the periplasm of this strain allows them to be functionally effective while membrane stability is maintained.

Biochemical-Morphological Modifications of Platelet Membranes during Pregnancy-Induced Hypertension

A platelet (PLT) function modification has been reported in normal pregnancies compared with the nonpregnant condition and it has been hypothesized to play a central role in the pathogenesis of pregnancy-induced hypertension (PIH). The aims of the present study were (i) to evaluate the lipid composition, fluidity at different depths, transport functions, and ultrastructural features of the PLT membrane in PIH and (ii) to ascertain whether similar modifications may be determined by the state of pregnancy in comparison with the nonpregnant condition. The platelets of healthy pregnant women (HPW) showed: (i) an increase in Ca 2+ ATPase activity, (ii) a decreased fluidity of the deeper site of the membrane, (iii) a reduced cholesterol concentration, with an increased ratio between unsaturated and saturated fatty acids; (iv) a decreased intramembranous particles (IMP) distribution factor (DF) of the plasma membrane E face in comparison with healthy nonpregnant women. When comparing women affected by PIH with HPW, we observed (i) reduced Na +/K + ATPase activity and enhanced Ca 2+ ATPase activity and intraplatelet calcium concentrations, (ii) an increased membrane fluidity, (iii) an increased cholesterol concentration and ratio between unsaturated and saturated fatty acids, (iv) a reduction of the IMP number and the DF. Pregnancy produces a deep modification of the platelet plasma membrane, and PIH produces a more pronounced alteration of the maternal platelets, which can be responsible for the observed modifications in placental blood flow and in the fetomaternal exchange.

Giardia lamblia: freeze-fracture ultrastructure of the ventral disc plasma membrane.

The close contact of Giardia lamblia trophozoites with mucosal surfaces produces surface indentations of epithelial cells, that progress to areas of microvilli depletion. This interaction is mediated by the lateral crest, a specialized contractile structure in the ventral disc of the parasite. We have analyzed the plasma membrane of the ventral disc using freeze fracture electron microscopy to study the distribution of large integral proteins, and freeze fracture cytochemistry with filipin to reveal sterol-containing sites. A previously undetected structural specialization was found at the lateral crest as a horseshoe-shaped membrane domain characterized by a near absence of intramembrane particles, in sharp contrast to the remainder of the ventral disc plasma membrane. In addition, the lateral crest showed a striking paucity of cholesterol-containing complexes in replicas of trophozoites treated with filipin. These observations demonstrate that the plasma membrane of G. lamblia displays a microheterogeneity in the planar distribution of cholesterol and intramembrane particles. Both membrane components are noticeably less abundant at the outer rim of the ventral disc where contraction takes place.

Cytological characterization of isolated sperm cells of perennial ryegrass (Lolium perenne L.)

The cytoplasmic content and the distribution of intramembrane particles (IMPs) of the plasma membrane of isolated sperm cells of perennial ryegrass (Lolium perenne L.) have been characterized using flow cytometry, transmission electron microscopy, confocal scanning laser microscopy and freeze-fracture studies. The isolated haploid sperm cells contain a variety of cell organelles with the exception of microtubules. Proplastids and plastids with starch were observed, although only rarely. Vacuoles containing remnants of organelles and stacked lamellae of endoplasmic reticulum with cytoplasmic inclusions were observed frequently, indicating that autophagy takes place. The number of mitochondria varies from 11 to 26 with an average of 17. Generally, the nucleus has a lobed shape and displays various interphasic stages of chromatin condensation. The analysis of the number of mitochondria and the nuclear state did not show evidence of sperm cell dimorphism. The cytological variability observed, could be explained by differences in developmental stages already present in vivo at the moment of isolation. No correlation between the number of mitochondria and the nuclear cross-sectioned area and/or the condensation state of the chromatin could be found. The density of intramembrane particles of the plasma membrane on the exoplasmic fracture face is more than twice that on the protoplasmic fracture face. That is the opposite of what was found for sporophytic cells of perennial ryegrass. These results are discussed in relation to the potential use of these cells for biotechnology and developmental studies.

Treponema pallidum rare outer membrane proteins: analysis of mobility by freeze-fracture electron microscopy.

Freeze-fracture and deep-etch electron microscopy were used to investigate the molecular architecture of the Treponema pallidum outer membrane (OM). Freeze-fracture electron microscopy of treponemes freshly harvested from rabbit testes revealed that the intramembranous particles (IMPs) in both the concave and convex OM leaflets were distributed into alternating areas of relatively high and low particle density; in many OM fractures, IMPs formed rows that ran either parallel to or obliquely across the fracture faces. Statistical analysis (runs test) confirmed that the IMPs were nonrandomly distributed in both OM leaflets. Examination of deep-etched specimens revealed that the particles observed in freeze-fractured OMs also were surface exposed. Combined analysis of deep-etched and cross-fractured treponemes revealed that the OM particles were located in regions of the OM away from the endoflagella and closely apposed to the cytoplasmic membrane-peptidoglycan complex. When treponemes were incubated for extended periods with heat-inactivated immune rabbit syphilitic serum, no alteration in the distribution of OM IMPs was detected. In further experiments, approximately 1:1 mixtures of T. pallidum and Escherichia coli or separate suspensions of the nonpathogenic Treponema phagedenis biotype Reiter were fixed at 34 degrees C or after cooling to 0 degree C (to induce lateral phase separations that would aggregate IMPs). Only particles in the T. pallidum OM failed to aggregate in cells fixed at the lower temperature. The combined data suggest that the mobility of T. pallidum rare OM proteins is limited, perhaps as a result of interactions between their periplasmic domains and components of the peptidoglycan-cytoplasmic membrane complex.

Electrical and Freeze-Fracture Analysis of the Effects of Ionic Cadmium on Cell Membranes of Human Proximal Tubule Cells

We previously reported that cell cultures of human proximal tubule (HPT) cells respond to ionic cadmium in a manner consistent with well-defined Cd(2+)-elicited responses reported for in vivo systems. However, one unique finding was that the transepithelial electrical resistance and tight junction sealing strands were altered as a result of Cd2+ exposure at micromolar concentrations. These alterations are reexamined in detail in the present report to determine whether the Cd(2+)-induced alterations are specific alterations in the tight junction structure or reflect a general alteration in the cell membrane. Exhaustive analysis of tight junction sealing strands demonstrated no significant alterations due to Cd2+ exposure, even at the concentration that elicited a significant reduction in transepithelial resistance. Further analysis of intramembrane particle distribution demonstrated a significant increase in apical intramembrane particles, indicating that Cd2+ exposure altered the characteristics of the apical cell membrane. Overall, the results were consistent with evidence of Cd(2+)-induced alteration in the apical cell membrane of the HPT cell.

Electrical and freeze-fracture analysis of the effects of ionic cadmium on cell membranes of human proximal tubule cells.

We previously reported that cell cultures of human proximal tubule (HPT) cells respond to ionic cadmium in a manner consistent with well-defined Cd(2+)-elicited responses reported for in vivo systems. However, one unique finding was that the transepithelial electrical resistance and tight junction sealing strands were altered as a result of Cd2+ exposure at micromolar concentrations. These alterations are reexamined in detail in the present report to determine whether the Cd(2+)-induced alterations are specific alterations in the tight junction structure or reflect a general alteration in the cell membrane. Exhaustive analysis of tight junction sealing strands demonstrated no significant alterations due to Cd2+ exposure, even at the concentration that elicited a significant reduction in transepithelial resistance. Further analysis of intramembrane particle distribution demonstrated a significant increase in apical intramembrane particles, indicating that Cd2+ exposure altered the characteristics of the apical cell membrane. Overall, the results were consistent with evidence of Cd(2+)-induced alteration in the apical cell membrane of the HPT cell.

Sperm morphology and distribution of intramembranous particles in the sperm heads of selected freshwater teleosts

The morphology of spermatozoa and the distribution of intramembranous particles (IMPs) in sperm-head membranes in teleostean fish were examined ultrastructurally to clarify the presence of characteristic arrays (parallelogram or hexagon in packing) of IMPs. The following four species of fish were used: goldfish ( Carassius auratus), loach ( Misgurnus anguillicaudatus), flat bitterling ( Acheilognatus rhombeus), sweetfish ( Plecoglossus altivelis). It was demonstrated that spermatozoa of all these fish were devoid of an acrosomal structure in the anterior portion of the head. Spermatozoa had round heads in goldfish, loach and flat bitterling. Two centrioles (proximal centriole and basal body) were present and located adjacent to each other in all fish. The characteristic arrays of IMPs were found in spermatozoa of goldfish and flat bitterling. IMPs were more numerous on the P-face than on the E-face in all species. The present work showed that the characteristic arrays of IMPs were not common structures in spermatozoa of teleostean fish.