Histochemical and ultrastructural characterisation of mantle storage cells in the hydrothermal-vent bivalve Bathymodiolus azoricus

This study reports on two types of storage cells that are present in the mantle connective tissue of the deep-sea mussel Bathymodiolus azoricus. One type corresponds to the adipogranular cells, a kind of storage cell previously described in other bivalves. In these cells extensive regions of the cytoplasm are filled with glycogen deposits and these zones became strongly stained after histochemical (PAS) or ultrastructural detection of polysaccharides. Several lipid droplets and membrane bound granules containing homogeneous electron-dense material are also present in adipogranular cells. A second type of cell contains large lysosomes in addition to numerous lipid droplets, but lacking cytoplasmic glycogen deposits. Due to these characteristics we named them adipolysosomal cells. They can be identified in semi-thin sections stained with PAS reaction because the lysosomes are the only positively stained structures. In the connective tissue of the mantle, some cells containing many lysosomes and a few lipid droplets were also observed. These cells differ from the adipolysosomal cells mainly because they have a reduced amount of lipid reserves, and could be an initial stage in the development of adipolysosomal cells. The vesicular connective tissue cells that in other Mytilidae are specialised in glycogen storage were not detected in B. azoricus. The reserves accumulated in the two types of storage cells described in B. azoricus may be important for the survival of these hydrothermal-vent bivalves if their nutrition is affected by a temporary loss or reduction of endosymbiotic bacteria due to sulphide and/or methane shortage caused by oscillations in vent activity.     

Oogenesis in the marine mussel Mytilus edulis: an ultrastructural study

Ultrastructural changes occurring during the course of development in oocytes of Mytilus edulis are described for mussels collected at monthly intervals over a period of one year (September 1981 to October 1982) from a site in Cornwall, England. During early stages of oogenesis the oocyte is surrounded by a small number of follicle cells but, as development proceeds, the follicle cells are restricted to the stalk region which attaches the oocyte to the acinar wall. Contact between the follicle cells and the developing oocyte is maintained by means of desmosomelike gap junctions. Organelles and inclusion bodies present in the ooplasm during oogenesis include rough endoplasmic reticulum (RER), Golgi bodies, mitochondria, free ribosomes, Balbiani's vitelline body, annulate lamellae and yolk and cortical granules. The RER, in particular, varies considerably throughout the course of development. Evidence for uptake of exogenous macromolecules into oocytes by pinocytosis is presented; it occurs in the basal region of previtellogenic oocytes prior to the formation of the vittelline coat. Lipid-yolk granules invariably have mitochondria in close association and, during the winter months, develop in close proximity to small, apparently glycogen-rich vesicles possibly suggesting that conversion of glycogen to lipid takes place in developing oocytes. Oocyte degeneration was commonly observed and involves initial breakdown of the plasma membrane followed by rupture of the vitelline coat. The oocyte contents once released into the acinar lumen are resorbed by the epithelial cells of the gonoducts, which are prevalent throughout the mantle of ripe individuals.     

Seasonal cycle of lysosomal enzyme activities in the mantle tissue and isolated cells from the musselMytilus edulis

InMytilus edulis L., gametogenesis takes place in the mantle at the expense of the connective storage tissue. There are two main types of storage cells: vesicular (VC) cells storing large amounts of glycogen and adipogranular (ADG) cells containing large numbers of protein granules, lipid droplets and lesser amounts of glycogen. One of the ways in which stored reserves can be mobilized for gamete formation is by controlled autophagy, in which the cellular constituents are degraded by lysosomes. Mussels were collected from the Menai Strait, North Wales, and monthly measurements made, over two years (1984–1986), of the activities of lysosomal acid hydrolases (acid phosphatase,-N-acetyl-D-glucosaminidase and-glucuronidase) and Cathepsins B and L in the mantle tissue, isolated ADG cells, low-density cells and, during spawning, in the mature oocytes of female mussels. The lysosomal proteinases, Cathepsins B, L and H, were further characterised by activation with thiol compounds and inhibition with thiol blockers and by leupeptin. Because of the low activity in the mantle tissue ofM. edulis, Cathepsin H was not assayed on a seasonal basis. There was a general increase in lysosomal enzyme activity during the winter, which can be related to increased autolysis in the storage cells and to the process of maturation in the developing oocytes. The activity of Cathepsin B was highest in the ADG and low-density cells, implying an important role in proteolysis within the ADG cells. By contrast, Cathepsin L displayed the highest activity in the mature oocytes, suggesting a major function of Cathepsin L in the development and maturation of the oocytes. Two different-glucosidase activities were measured in the monthly assays, one with a pH optimum of 4.5 (acid) and the other at pH 7.5 (neutral). Highest activities of the acid-glycosidase were found in the low-density cells, but there were no significant seasonal changes in the mantle tissue as a whole. Activities of the neutral-glucosidase were low in the ADG cells and mature oocytes, but showed high activities in the mantle tissue, with marked seasonal changes that corresponded to the mobilization of glycogen reserves in the VC cells.     

Studies on digestion and the fine structure of digestive caeca in Eurydice pulchra (Crustacea: Isopoda)

In the carnivorous isopod Eurydice pulchra Leach the mid-gut caeca have a pH of 5.6 and produce a strong acid proteinase, a lipase and a carbohydrase. Electron microscope studies of the digestive caeca of starved and fed animals show that secretion is merocrine and may originate from a complex involving the basal cell membrane and associated mitochondria, producing lysosomes. Absorbed materials, in fed animals, appear to take the form of neutral lipid bodies, glycogen, and proteinaceous crystals, the latter forming within the golgi complex. Digestion of both lipid and protein inclusions was observed to occur intra-cellularly by fusion with free lysosomes. Secretion and absorption may occur in the same cell, and the scheme for the sequence of epithelial cell stages is proposed.     

Ultrastructure of the gonad and gametogenesis in the eastern oyster,Crassostrea virginica. I. Ovary and oogenesis

The ultrastructural features of the ovary and oogenesis are described in the eastern oyster,Crassostrea virginica (Gmelin, 1791). The ovary is a diffuse organ consisting of highly branching acini in which oocytes develop. The acini are surrounded by a matrix of vesicular connective tissue (VCT tells) which serves a nutrient storage function. Each acinus is bathed by fluid within a surrounding connective tissue compartment, the hemocoel, which likely serves as a means of transporting nutrients to the oocytes. Oocytes begin growth while positioned near to the inner acinus wall. As differentiation proceeds and they enter the late stages of vitellogenesis, they become stalle-shaped and project into the acinus lumen. Follicle tells are closely associated with oocytes during the early and middle stages of vitellogenesis but they are largely confined to the basal, stalked region of late-stage oocytes. Vitellogenesis occurs through a process of autosynthesis, involving the combined activity of the Golgi complex and rough endoplasmic reticulum, and heterosynthesis in which extraovarian precursors are incorporated into oocytes via receptor-mediated endocytosis involving the basal surface of the oocytes. It is suggested that the follicle tells play some important role during oogenesis but probably are not the major source of yolk precursors. The VCT celas are probably the main source of nutrients for vitellogenesis.     

Intracellular cyanobiont Richelia intracellularis: ultrastructure and immuno-localisation of phycoerythrin,nitrogenase, Rubisco and glutamine synthetase

In marine tropical or subtropical plankton the filamentous, heterocyst-forming cyanobacterium Richelia intracellularis forms a symbiosis with the diatom Rhizosolenia clevei. An ultrastructural analysis of the apex of Rhizosolenia clevei showed that the cytoplasm in that particular part of the cell was present only where the cyanobiont was located. The cyanobiont was, however, always outside the host cytoplasm. Vegetative cells as well as the heterocysts of the cyanobiont were devoid of gas vesicles and cyanophycin granules, while carboxysomes and large glycogen granules were common. The cyanobacterial cell wall apparently remained intact in both vegetative and heterocyst cells. In green excitation light the heterocysts and vegetative cells emitted a bright yellow fluorescence, indicating that both cell types possessed high concentrations of the pigment phycoerythrin (PE) commonly associated with photosystem (PS) II. The presence of this pigment in both cell types was verified by immunogold localisation. Using the same technique, the nitrogenase (dinitrogenase reductase) enzyme was shown to be exclusively present in the heterocysts, while Rubisco was localised primarily to the carboxy-somes, which were only detected in vegetative cells. Using an antiserum against the ammonia assimilating enzyme glutamine synthetase (GS), we could demonstrate very low levels of this enzyme, indicating repression of GS in the cyanobiont.     

An electron-microscope study of periostracum formation in some marine bivalves. II. The cells lining the periostracal groove

The histology and ultrastructure of the epithelial cells of the outer and middle mantle folds of the bivalves Mytilus edulis (L), Cardium edule (L), Macoma balthica (L) and Nucula sulcata Bronn are described. The cells lining the inner face of the outer fold exhibit a prominent granular endoplasmic reticulum, and Golgi complexes which are concerned with the elaboration of granules and vesicles eventually incorporated into the periostracum. A gradual reduction in the protein synthetic apparatus occurs towards the tip of the fold. Within the cells, it is proposed that the ovoid inclusion bodies are lysosomes and that they control the rate of secretion. The cells of the middle fold are cuboidal in appearance. Those of M. edulis and N. sulcata exhibit prominent granules, whereas those of C. edule and M. balthica possess vesicles. The cells of M. edulis differ from the others in possessing stout bundles of filaments, which occupy large areas of the cell and constitute a cell web. The cells of the epithelium in all cases do not appear to be implicated in periostracum formation.     

Microarchitecture and function of the lophophore in the bryozoan Cryptosula pallasiana

The architecture and function of the lophophore of the marine bryozoan Cryptosula pallasiana (Moll) are described, including some new features not previously discovered in bryozoans. The nature of fluid movements within the lophophoral coelom during feeding activities is postulated on the basis of the arrangements of epithelia and muscles. Epithelial cells at the tentacle bases are blastemic in nature, and there is a ciliated pit of unknown function in the angle between every pair of tentacles. There are 6 nerves in each tentacle, including a pair of single-axon subperitoneal nerves. Neurosecretory-like vesicles and glycogen occur in some neurons of the ganglion. The basal lamina collagen has a diameter smaller than that previously recorded for an invertebrate. Filament dimensions are given for several different muscles. Tentacle muscles and lophophore retractor muscles are smooth. Thick paramyosin-like filaments up to 75 nm diameter occur in two muscle types. A new set of muscles is described: the basal transverse muscles of the tentacles.     

6-HO-BDE-137对大鼠睾丸支持细胞的毒性

以原代培养的大鼠睾丸支持细胞为研究对象,选取环境及生物体中均有检出的一种典型的羟基化PBDE——6-HO-BDE-137作为目标化合物,设置0、0.1、1、10μmol·L-14个浓度梯度,应用噻唑蓝(MTT)试验、荧光显微镜观察和AnnexinV-FITC/PI双染流式细胞术,通过检测大鼠睾丸支持细胞增殖活性、细胞形态和细胞凋亡坏死率的变化,探讨其对支持细胞的损伤情况.研究发现,6-HO-BDE-137显著影响支持细胞的增殖活力,暴露24h,10μmol·L-1浓度组细胞与对照组相比显著增殖(p<0.05),随暴露时间的延长(至48h),10μmol·L-1组转为增殖抑制;支持细胞形态也表现出不同程度的改变,10μmol·L-1浓度组变化最为明显,有大量的细胞变圆飘起;随6-HO-BDE-137暴露浓度的升高,支持细胞死亡率增加,凋亡是支持细胞死亡的主要方式.研究结果提示:6-HO-BDE-137具有生殖毒性.     

Potential histopathological and molecular changes in rat vas deferens inhaled by Boswellia papyrifera and Boswellia carterii

Boswellia papyrifera and Boswellia carterii released from smoke contaminate indoor environment and consequently adversely affect humans as evidenced by respiratory disturbances. The aim of this study was to determine the effects of these plants on pathological and biochemical changes in vas deferens of albino rats. Animals were administered 4 g/kg body weight B. papyrifera and B. carterii daily for 120 days along with controls. Significant changes were observed in epithelial cell types and some cells showed signs of degeneration. The ultrastructural studies revealed marked changes in cytoplasmic organelles. Microvilli were missing and lysosomes were found in the cytoplasm. In addition, all treated groups plasma fructose and other biochemical parameters were decreased indicating reduced energy necessary for motility and contractility of spermatozoa. Many spermatozoa were disorganized and agglomerated. Data suggest that smoke from these plants adversely affects vas deferens.     

Oocyte development in the kuruma prawn Penaeus japonicus

Female kuruma prawns (Penaeus japonicus Bate) with undeveloped, early developing, developing, nearly ripe and ripe ovaries, were collected from Ise Bay, Japan, in 1984. Oocyte development of the kuruma prawn was classified into ten stages according to morphological characters, namely: (1) synapsis stage, (2) chromatin nucleolus stage, (3) early perinucleolus stage, (4) late perinucleolus stage, (5) oil globule Stage I, (6) oil globule Stage II, (7) yolkless stage, (8) yolk granule stage, (9) prematuration stage, and (10) maturation stage. The synapsis stage is a multiplication stage. The chromatin nucleolus stage, early and late perinucleolus stages are previtellogenesis and primary growth stages. Oil globule Stage I is an initial stage of primary vitellogenesis and secondary growth. Follicle cells on the oil globule Stage I oocytes expand rapidly and reach maximum size during oogenesis. Yolk granule stage oocytes are in the initial stages of secondary vitellogenesis. Strongly acidophilic yolk granules accumulate within basophilic vesicles of the cytoplasm. The yolk granules are first concentrated in the inner part of the cytoplasm, then gradually spread to the periphery. Cortical crypts, which are separated from the oocyte cytoplasm by the cytoplasmic membrane, are situated outside of oocyte cytoplasm. Germinal vesicle breakdown (GVBD) is initiated in the late phase of prematuration and continues until the late phase of maturation immediately prior to spawning. At the beginning of the maturation stage, the oocytes are ovulated, after which the nuclei further shrink and migrate out-wards. After ovulation, meiotic division of the ovarian oocyte progressed up to the metaphase of primary maturation division. Finally, the meiotic metaphase is visible just beneath the cytoplasmic membrane in the mature oocyte. Though ovulation is synchronous within the same ovary, GVBD is not completely synchronous. Ovulated mature oocytes have many club-shaped cortical crypts in the peripheral part of the cytoplasm and contain extensive accumulations of yolk granules dispersed throughout the cytoplasm. The apical end of the club-shaped cortical crypts and cytoplasmic membrane are coated by the vitellin envelope in the mature oocyte.     

An electron-microscope investigation of the cells lining the outer surface of the mantle in some marine molluscs

The ultrastructural details of the cells lining the outer surface of the mantle in Mytilus edulis (L), Cardium edule (L), and Nucula sulcata Bronn. are described. Along the length of the outer mantle fold and the general outer surface of the mantle, the epithelial cells differ, with a progressive reduction in the protein synthetic apparatus and an increase in glycogen, while large numbers of mitochondria are evident in the apical and basal regions.     

镉诱导仔猪睾丸支持细胞氧化酶活性降低及DNA损伤的研究

以仔猪睾丸支持细胞为实验模型,采用二步酶消化法分离支持细胞进行培养。探讨了0、10、20、40、80gmol·L^-1的氯化镉对支持细胞的毒性作用。结果表明：10gmol·L^-1以上的氯化镉有抑制支持细胞生长的作用,并能使支持细胞氧化酶活性下降,造成支持细胞DNA损伤。     

Cu2+或表面活性剂AE对黄鳝肝损伤的超微结构观察

利用透射电镜研究了亚致死浓度的Cu^2 或表面活性剂AE(脂肪醇聚氧乙烯醚)对黄鳝肝细胞结构的损伤作用．观察发现，在ρ(Cu^2 )=4．5mg／L的污染环境中，黄鳝肝脏细胞核变形，染色质凝集，内质网囊泡化，细胞质中出现较多的溶酶体和过氧化物酶体；在ρ(AE)=4．0mg／L的表面活性剂污染下，黄鳝肝脏细胞核变形，内质网呈线状，溶酶体、过氧化物酶体体积大、数量多，部分细胞质解体，细胞中出现空腔甚至死亡．图版1参10     

Annual cycle of expression of connective tissue polypeptide markers in the mantle of the musselMytilus galloprovincialis

We suggest that gonad development in the mantle tissue of the bivalve molluscMytilus galloprovincialis Lmk. is an example of epithelial/mesenchymal interactions (i.e. soma/germline interactions) and morphogenesis in the adult state. According to this concept, the aim of the present study was to use biochemical and immunochemical methods for identifying and characterizing the mantle cell polypeptide markers whose expression is seasonally and morphogenetically regulated. We showed for the first time thatM. galloprovincialis mantle, of both males and females, contains polypeptides (with an apparent mol. wt of 45 to 53 kDa) specific for connective tissue (mantle connective tissue polypeptides; MCTPs). Electrophoretic, immunoblotting and immunofluorescent experiments demonstrated that MCTPs are primarily localized in the adipogranular (ADG) cells, and their expression in the mantle is seasonally regulated. There is a positive correlation between MCTP expression and connective tissue volume in the mantle. MCTPs are overexpressed during the rest period, when the mantle consists of connective tissue mainly, whereas mature gonads contain only trace amounts of MCTPs. Moreover, there is a temporal correlation between the onset and decrease of MCTP expression and the appearance and disappearance of the ADG cells in the mantle. MCTP localization in the mantle tissue should not be associated with the ADG cells only, because positive immunofluorescence was also detected in follicle membranes (but not in germ cells) and superficial mantle epithelium. Using immunoblotting and immunofluorescence, MCTPs were found in the connective tissue of the mantle, posterior adductor muscle and visceral mass, but not gills, foot or hepatopancreas. Possible mechanisms by which MCTPs could participate in the annual processes of mantle gonad/connective tissue development and involution are discussed.     

Ultrastructure changes in hepatocytes of catfish Clarias gariepinus from Lake Mariut, Egypt

In the present study, specimens of catfish (Clariidae) were collected from a polluted location (Main Basin) and a relatively clean area (East Basin) in Lake Mariut, one of the Nile Delta Lakes in Egypt. Fifteen fish were taken from each site. Liver preparations of fish from the two sources were comparatively examined for cellular changes using transmission electron microscopy. Fish hepatocytes from the polluted area showed accumulation of the heterochromatin, enlarged nucleoli, and an extremely folded nuclear envelope. Perichromatin granules were increased and progressively formed small clusters closely associated with patches of heterochromatin. In the cytoplasm, fractionation, dilation, and vesiculation of rough endoplasmic reticulum (RER), and elevated amounts of smooth endoplasmic reticulum (SER) tubules were noted. The most frequent pathological modifications were the swelling of mitochondria, cristae regression and changes in the electron-transparency of the matrix. Lysosomes showing myelin-like stacks of membraneous material (phospholipidosis), glycogenosomes (i.e., glycogen rosettes enclosed by membranes) and cytoplasmic myelinated bodies were strongly developed. Furthermore, increasing numbers of secondary lysosomes with degraded cell organelles were found. With reference to the storage vesicles, there appeared to be an increase in the lipid droplets (lipidosis) within many hepatocytes. This study reinforces the need to select representative sentinel species from different habitats for biomonitoring purposes and it provides further support for the use of biomarkers in assessing the health of aquatic ecosystems.     

Electron microscopy documenting the cellular metabolic fate of Zn IONS∗

In vertebrates and invertebrates Zn exist as complexed compounds with metallothioneins. However, its cellular level effects and metabolic fates are scantly documented. In the elucidation of this fact, EM results on hepatic cellular alterations in fish under lethal dose exposure at 4.0 ppm over a week is illustrated.

A large number of lysosomes in hepatic cells prevailed on exposure to Zn in its sulfate form. Evidently, due to metal compound stress and cellular damage lysosomal activity is augmented. The lysosomes harboured digestible material, presumably the aforementioned substrates. Contrary to it, fat droplets prevailed while glycogen depletion is noticeable. Unlike the effects of Hg, the nuclei were normal with granular chromatin and prominent nucleoli. However, the mitochondria contained some small intramitochondrial bodies. Similar to the effects of Hg, the cell membrane remained intact.

In vivo enzymatic studies indicated augmentation in catalase, acid‐ and alkaline‐phosphatases, while glucose‐6‐phosphatase is inhibited. However, only alkaline‐ and glucose‐6‐phosphatases are inhibited under in vitro conditions.

Thus, it is evident that Zn enhances cellular bioenergetic requirements culminating in glycogen depletion owing to stress, concomitantly envisaging inhibition of oxidative phosphorylation in the electron transport system.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Fine structure of naive and allogeneic challenged ampullae in Botrylloides subpopulation I from the Mediterranean coast of Israel

The external peripheral termini of vasculature in botryllid ascidians (ampullae) are important in the expression of effector mechanisms during histocompatibility reactions. We study here morphological changes to the microstructure during allorecognition in Botrylloides subpopulation I (SP1) from the Mediterranean coast of Israel, where special giant ampullae, called megaloampullae are developed hours after first allogeneic challenge. The basal part of each naive ampulla consists of a sheath of continuous squamous epithelial cells (1 μm×20 μm). At the tip of the ampullae, about 130 epithelial cells become columnar with wide apical pads (15 μm×5–10 μm), that contain electron-opaque “adhesive vesicles”. Foci of crystallizations were observed within the vesicles of some columnar cells. Ampullar epithelial cells develop internal folds that support tight attachments between circulating blood cells (most of them are pigment cells) and ampullar epithelium. During the histoincompatibility process, the tunic peripheral edge at the interaction area becomes highly convoluted. Filaments (up to 1.16 μm) accumulate in the interaction tunic matrix. Some of these filaments encircle destructed cells. Fragmented cell membranes are also found next to the reacting ampullae. The most characteristic feature of the Botrylloides SP1 rejection reaction is dilation of the ampullae. The ampullar epithelium becomes “curly” when megaloampullae are formed. Within the curly region, about 30% of the squamous epithelial cells are further stretched, up to 33 μm in length. Some additional mechanisms are suggested to explain the fast increase (up to seven times) in ampullar width that is recorded within a few hours from the first allogeneic contact.     

Spermatogenesis,sperm storage and comparative sperm morphology in nine species of Capitella,Capitomastus and Capitellides (Polychaeta: Capitellidae)

Light microscopy and transmission electron microscopy (TEM) were used to describe spermatogenesis and the morphology of mature sperm and sperm storage organs in five sibling species of Capitella, three species in the related genus Capitomastus, and one species in the genus Capitellides. These capitellids lack a well-developed testis, but young males have a few specialized regions of the peritoneum in the eighth setiger, where germ cells proliferate and spermatogonia are released into the coelom, and spermiogenesis is completed. Mature sperm are stored in the central regions of paired genital ducts (coelomoducts), which lie between the seventh and eighth setigers. The cells forming the walls of the coelomostome and central region of the duct are ciliated and have large glycogen deposits. The lumenal borders have extensive microvilli and there is evidence that they secrete glycogen-containing materials into the duct. All species have modified primitive sperm with a conical acrosome, elongated nucleus, and long middle piece extending along the proximal portion of the flagellum. A single ring-shaped mitochondrion encircles the centriolar region of the middle piece and the cytoplasm is filled with glycogen. The sperm of all nine species differ significantly in the lengths of their middle pieces, acrosomes and especially in their nuclear lengths. The nuclear lengths have a twofold range among the sibling species of Capitella and Capitomastus. Subtle differences in the shape and volume of the acrosomal vesicle and acrosomal space characteristic of the Capitella sibling species seem to correlate with a basic division of these species into those with diploid chromosome numbers of 20 or 26. Spermiogenesis, the number of sperm produced, and the method of sperm storage are appropriate for efficient sperm utilization in fertilization. No evidence indicates that spermatophores are formed and transferred between individuals and the method of sperm transfer is not understood. The differences in the dimensions and acrosome morphology of mature sperm, and the previously demonstrated specializations in the egg envelopes in the Capitella sibling species, are characteristic features of the reproductive isolation that exists among these capitellid species.