Components and development of the centriolar complex during and beyond spermiogenesis in a passeridan bird, the Masked weaver (Ploceus velatus)

The fate of the proximal centriole in passeridan birds is an area of controversy and relative lack of knowledge in avian spermatogenesis and spermatology. This study examines, for the first time, spatiotemporal changes in the centriolar complex in various phases of spermiogenesis in a passerine bird, the Masked weaver (Ploceus velatus). It also describes the configuration of the centriolar complex and the relationship between it and the granular body in both intra- and extra-testicular spermatozoa. It is shown that the proximal centriole is retained and attaches, at its free end, to the granular body of spermatids in every step of spermiogenesis, as well as in mature intra-testicular and post-testicular spermatozoa, including those in the lumen of the seminal glomus. As the centriolar complex, along with its attached granular body, approaches the nucleus in the early spermatid, the proximal centriole articulates with the distal centriole at an acute angle of about 45°, and thereafter, both centrioles, still maintaining this conformation, implant, by means of their articulating proximal ends, at the implantation fossa of the nucleus. In the mature spermatid and spermatozoon, the granular body winds itself helically around the centriolar complex in the neck/midpiece region of the cell, and, thus, becomes the granular helix. The significance of this observation must await future studies, including possible phylogenetic re-evaluation and classification of birds.     

Immunohistochemistry of the cytoskeleton in the excurrent ducts of the testis in birds of the Galloanserae monophyly

The presence, location and degree of immunoexpression of various microfilament (MF) and intermediate filament (IF) systems (actin, cytokeratins, desmin, vimentin) were studied in the excurrent ducts of the testis in sexually mature and active galliform (Japanese quail, domestic fowl, turkey) and anseriform (duck) birds. These proteins were variably expressed between the epithelia and periductal tissue (periductal smooth muscle cell layer and interductal connective tissue) types and between species. Variable heterogeneous co-expression of filament systems was also found in the various duct epithelia and periductal tissue types: co-expression of filament systems was the rule rather than the exception. In the duck, neither vimentin nor cytokeratin was present in any of the tissues, whereas actin and desmin (absent in the rete testis) were co-expressed in the efferent ducts and epididymal duct unit (comprising the ductus conjugens, ductus epididymidis and ductus deferens). Actin, desmin and vimentin were generally co-expressed in the rete testis, efferent ducts and epididymal duct unit of the quail, domestic fowl and turkey, with vimentin being more strongly immunoreactive than actin and desmin in the epididymal duct unit, but more weakly immunoexpressed in the efferent ducts. Cytokeratin was present and co-expressed with actin, desmin and vimentin in the rete testis, efferent ducts and epididymal duct unit of the domestic fowl and turkey, but not in the quail and duck. The periductal smooth muscle cell layer and interductal tissue co-expressed actin, desmin and vimentin variably in all birds. Luminal spermatozoa of both the turkey and duck were immunonegative for all protein systems, whereas those of the quail and domestic fowl co-expressed actin, desmin and vimentin moderately or strongly. The tissues of the reproductive tract of male birds thus contain cytoskeletal protein systems that are variably but mostly co-expressed and whose contractile ability appears necessary and sufficient for transportation through the various excurrent ducts of the voluminous testicular fluid and its high sperm content, characteristic features of male avian reproduction.     

Immunolocalization of aquaporins 1, 2 and 7 in rete testis, efferent ducts, epididymis and vas deferens of adult dog

The transepithelial movement of water into the male reproductive tract is an essential process for normal male fertility. Protein water channels, referred to as aquaporins (AQPs), are involved in increasing the osmotic permeability of membranes. This study has examined the expression of AQP1, AQP2, and AQP7 in epithelial cells in adult dog efferent ducts, epididymis, and vas deferens. Samples of dog male reproductive tract comprising fragments of the testis, initial segment, caput, corpus and cauda epididymidis, and vas deferens were investigated by immunohistochemistry and Western blotting procedures to show the localization and distribution of the AQPs. AQP1 was noted in rete testis, in efferent ducts, and in vessels in the intertubular space, suggesting that AQP1 participated in the absorption of the large amount of testicular fluid occurring characteristically in the efferent ducts. AQP2 expression was found in the rete testis, efferent ducts and epididymis, whereas AQP7 was expressed in the epithelium of the proximal regions of the epididymis and in the vas deferens. This is the first time that AQP2 and AQP7 have been observed in these regions of mammalian excurrent ducts, but their functional role in the dog male reproductive tract remains unknown. Investigations of AQP biology could be relevant for clinical studies of the male reproductive tract and to technologies for assisted procreation. R.F.D. gratefully acknowledges a Fellowship from the Department of Anatomy, Institute of Biosciences, UNESP, Botucatu, SP, Brazil. This work was also funded by FAPESP (Sao Paulo State Research Foundation; grant 04/05578–1 to A.M.O. and grant 04/05579–8 to R.F.D.). This paper is part of the PhD Thesis presented by R.F.D. to the State University of Campinas – UNICAMP, Brazil.     

Morphology and immunolocalization of aquaporins 1 and 9 in the agouti (Dasyprocta azarae) testis excurrent ducts

This study investigated the morphology and immunoexpression of aquaporins (AQPs) 1 and 9 in the rete testis, efferent ducts, epididymis, and vas deferens in the Azara’s agouti (Dasyprocta azarae). For this purpose, ten adult sexually mature animals were used in histologic and immunohistochemical analyses. The Azara’s agouti rete testis was labyrinthine and lined with simple cubic epithelium. Ciliated and non-ciliated cells were observed in the epithelium of the efferent ducts. The epididymal cellular population was composed of principal, basal, apical, clear, narrow, and halo cells. The epithelium lining of vas deferens was composed of the principal and basal cells. AQPs 1 and 9 were not expressed in the rete testis. Positive reaction to AQP1 was observed at the luminal border of non-ciliated cells of the efferent ducts, and in the peritubular stroma and blood vessels in the epididymis, and vas deferens. AQP9 was immunolocalized in the epithelial cells in the efferent ducts, epididymis and vas deferens. The morphology of Azara’s agouti testis excurrent ducts is similar to that reported for other rodents such as Cuniculus paca. The immunolocalization results of the AQPs suggest that the expression of AQPs is species-specific due to differences in localization and expression when compared to studies in other mammals species. The knowledge about the expression of AQPs in Azara’s agouti testis excurrent ducts is essential to support future reproductive studies on this animal, since previous studies show that AQPs may be biomarkers of male fertility and infertility.     

Duplicated growth hormone genes in a passerine bird, the jungle crow (Corvus macrorhynchos)

Molecular cloning, molecular phylogeny, gene structure and expression analyses of growth hormone (GH) were performed in a passerine bird, the jungle crow (Corvus macrorhynchos). Unexpectedly, duplicated GH cDNA and genes were identified and designated as GH1A and GH1B. In silico analyses identified the zebra finch orthologs. Both GH genes encode 217 amino acid residues and consist of five exons and four introns, spanning 5.2 kbp in GH1A and 4.2 kbp in GH1B. Predicted GH proteins of the jungle crow and zebra finch contain four conserved cysteine residues, suggesting duplicated GH genes are functional. Molecular phylogenetic analysis revealed that duplication of GH genes occur after divergence of the passerine lineage from the other avian orders as has been suggested from partial genomic DNA sequences of passerine GH genes. RT-PCR analyses confirmed expression of GH1A and GH1B in the pituitary gland. In addition, GH1A gene is expressed in all the tissues examined. However, expression of GH1B is confined to several brain areas and blood cells. These results indicate that the regulatory mechanisms of duplicated GH genes are different and that duplicated GH genes exert both endocrine and autocrine/paracrine functions.     

Localization of epidermal growth factor (EGF) and its receptor (EGFR) during postnatal testis development in the alpaca (Lama pacos)

The objective of the present studies was to determine the localization of epidermal growth factor (EGF) and the epidermal growth factor receptor (EGFR) in testicular tissue collected from male alpacas at 12 and 24 months of age. In the testes of 12-month-old alpacas, positive staining for EGF was not detected. EGFR was localized to Leydig cells within the 12-month-old alpaca testis, but staining was absent within seminiferous tubules. At 24 months of age, EGF was localized to Leydig cells, peritubular myoid cells, Sertoli cells and germ cells of the alpaca testis, with a preferential adluminal compartment staining within the seminiferous tubules. EGFR was also localized to the Leydig cells, peritubular myoid cells, Sertoli cells and germ cells within the 24-month-old alpaca testis, but staining within the tubules was primarily within the basal compartment. Results indicate distinct temporal and spatial regulation of EGF and EGFR in the alpaca testis and support a potential role for EGF and its related ligands in alpaca testis development and spermatogenesis.     

精索静脉曲张对青春期大鼠睾丸中VEGF和Flt-1蛋白表达的影响

目的研究血管内皮生长因子(VEGF)及其受体Flt-1蛋白在实验性左侧精索静脉曲张(ELV)大鼠睾丸中的表达和定位,探讨它们在精索静脉曲张(VC)致男性不育中的作用。方法建立青春期大鼠ELV模型,采用免疫组化法检测VEGF及Flt-1在ELV4周、8周组及相应对照组大鼠睾丸中的表达变化。结果 VEGF和Flt-1蛋白在大鼠睾丸中定位具有细胞特异性。VEGF蛋白表达于生精细胞、精子细胞发育中的顶体、Sertoli和Leydig细胞胞质内;Flt-1表达于精子细胞发育中的顶体及Leydig细胞胞质中。ELV4周组睾丸中VEGF蛋白的表达显著增加(P<0.01),8周时其表达量下降(P<0.01);ELV4周组与8周组睾丸中Flt-1蛋白的表达均比相应对照组下降(P<0.01),ELV8周组比4周组显著减少(P<0.01)。结论 ELV可影响青春期大鼠睾丸中VEGF和Flt-1蛋白的表达量,可能会影响精子的发生、发育,因而该变化可能是VC引起男性不育的原因之一。     

The identification and localization of two intermediate filament proteins in the tunic of Styela plicata (Tunicata, Styelidae)

The intermediate filament (IF) proteins Styela C and Styela D from the tunicate Styela (Urochordata) are co-expressed in all epidermal cells and they are thought to behave as type I and type II keratins. These two IF proteins, Styela C and Styela D, were identified in immunoblots of proteins isolated from the tunic of Styela plicata. The occurrence and distribution of these proteins within the tunic of this ascidian was examined by means of immunofluorescence and immunoperoxidase techniques, using anti-Styela C and anti-Styela D antibodies. In addition, immuno-electron microscopy of the tunic showed that the two proteins are located in the cuticle layer and in the tunic matrix. These results represent the first data about the presence of IF proteins in the tunic of adult ascidian S. plicata. The possible involvement of these IF proteins in reinforcing the integrity of the tunic, that represents the interface between the animal body and the external environment, is discussed.     

Development of the genital ducts in Telmatodrilinae (Tubificidae, Clitellata)

In Tubificidae, the male genital duct comprises a funnel in the testes segment, followed by a vas deferens, an atrium, and, frequently, a copulatory structure in the adjacent ovarian segment. There may also be a diffuse or compact prostate gland in association with the duct. The morphology and position of the genital ducts are important for the classification of the oligochaetous Clitellata. Different parts of the male duct, however, have been named without regard to whether they are homologous or not. One way to establish better hypotheses of homology is to study the detailed morphology and/or the development of the genital ducts. The morphogenesis of the genital ducts in Alexandrovia onegensis (Telmatodrilinae) is described. The male funnel originates by multiplication of peritoneal (mesodermal) cells in the posterior septum in the testes segment. A cord of these cells breaks through the septum and grows backwards into the next segment, where it connects to the epidermis. This cord gives rise to the vas deferens, and is therefore mesodermal in origin. The atrium in A. onegensis develops from a primary epidermal (ectodermal) invagination. The vas deferens and atrium connect and a continuous duct from the testes segment to the exterior is formed. Several compact prostate glands develop along the atrium, each being formed from cells in the atrial epithelium. The spermatheca develops from an invagination of the epidermis in the testes segment. The female duct is formed from peritoneal (mesodermal) cells in the posterior septum in the ovarian segment. These developmental findings strengthen the hypothesis about a closer relationship between the Telmatodrilinae and Tubificinae (both Tubificidae).     

Carotenoids, colour and conservation in an endangered passerine, the hihi or stitchbird (Notiomystis cincta)

Carotenoids are essential dietary components utilized not only in pigmentation but also as immuno-stimulants and antioxidants. Reduced availability can have consequences on individual health and survival, thus making carotenoids a good indicator of environmental stress. We compared carotenoid profiles and plumage colour characteristics of an endangered passerine species in New Zealand, between its remnant island source population and two reintroduced island populations. Circulating carotenoids were predominantly lutein (mean of 82.2%) and zeaxanthin (mean of 14.8%), and these were the major carotenoids present as yellow pigments in the males' plumage. There were clear differences in total carotenoid concentrations and plumage colour among the three populations. Circulating carotenoid concentration was significantly higher in one of the reintroduced populations, and the yellow plumage of males was significantly higher in both reintroduced populations in comparison with the remnant population (reflected as a significant increase in hue). Understanding how these differences arise may be of importance to this species given the health benefits carotenoids impart and our ability to select plant species containing these compounds or artificially supplement them.     

The Cytoskeleton of Primary Astrocytes in Culture Contains Actin, Glial Fibrillary Acidic Protein, and the Fibroblast-Type Filament Protein, Vimentin

Abstract: Primary astrocytes were cultured from the forebrains of 1-day-old rats. Immunofluorescence microscopy showed that approximately 80% of the cells were positive for glial fibrillary acidic protein (GFAP) and >80% were stained with an antiserum to the molecular weight 58,000 fibroblast intermediate filament protein (vimentin). Gel electrophoresis of Triton-insoluble cytoskeleton preparations from these cultures revealed three major bands having molecular weights of 58,000, 51,000, and 42,000, together with some prominent lower-molecular-weight species. The protein of molecular weight 51,000 was not present in preparations from fibroblasts. Each of the three major astrocyte proteins was subjected to limited proteolysis, while two of the proteins were cleaved by cyanogen bromide. The electrophoretic peptide patterns of the 58,000 protein were similar to those of vimentin isolated from NIL-8 fibroblasts, and the patterns of the 51,000 protein were similar to those of GFAP isolated from rat spinal cord. The patterns of the protein of molecular weight 42,000 resembled those of muscle actin. Rocket immunoelectrophoresis showed that the 51,000 astrocyte protein reacted with an antiserum to bovine GFAP, but the 58,000 and 42,000 proteins failed to react. We conclude that the major proteins of cytoskeleton preparations from cultured primary astrocytes are vimentin (58,000), GFAP (51,000), and actin (42,000), and that our data show no obvious structural relationship among them.     

The structure of Verson's cells in Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Testis follicles of Lepidoptera contain a large somatic cell termed Verson's cell. The present study focuses on the structure of Verson's cells and neighbouring germ cells in the Mediterranean mealmoth, Ephestia kuehniella (Pyralidae), using electron microscopy, antitubulin immunofluorescence, and phalloidin incubation for the visualization of microfilaments. Verson's cells of young larvae are connected with the follicle boundary and show large areas containing packages of glycogen particles, whereas Verson's cells of pupae lie freely within the testis follicle and are largely devoid of glycogen. Both developmental stages of Verson's cells have in common the presence of a dense cytoplasmic network of microtubules. A juxtanuclear subset of the cytoplasmic microtubule array is recognized by an antibody against acetylated microtubules. This indicates that more stable microtubules exist in this region. Microfilaments are arranged parallel to the cytoplasmic microtubules. The microtubule-microfilament-complex forms a cytoskeleton that may keep larger organelles, such as mitochondria and lysosomes, in a juxtanuclear position. Chromatin within the nuclei of Verson's cells is largely decondensed and nuclear pores are abundant. This indicates a high synthetic activity within the cells. The development of cells directly attached to Verson's cells, viz. prespermatogonia, may be controlled by the Verson's cells. Prespermatogonia, which differ in cytoplasmic density from spermatogonia further away from Verson's cells, may represent stem cells that give rise to spermatogonia and somatic cyst cells upon detachment from Verson's cells. This suggestion is compatible with the low division rate of prespermatogonia.     

Immunohistochemical localization of glial fibrillary acidic protein (GFAP) and vimentin in the subcommissural organ of the Mongolian gerbil (Meriones unguiculatus)

Summary The chemical composition of intermediate filaments (IF's) in the ependyma of the subcommissural organ (SCO) of the Mongolian gerbil (Meriones unguiculatus) was investigated immunohistochemically in paraffin-embedded tissue. Antibodies against glial fibrillary acidic protein (GFAP), vimentin, neurofilament proteins and cytokeratins were used. Only GFAP and vimentin were detected in the non-specialized diencephalic ependyma and in the ependymocytes of the SCO. Staining could be observed in apical and basal processes of the SCO-cells. The latter processes extended into the posterior commissure up to the subpial surface, thus establishing a well-developed leptomeningeal route of ependymal projections. In contrast to the homogeneous vimentin-labeling, the SCO was particularly immunoreactive for GFAP in its lateral aspects and in the supraand precommissural parts. The coexpression of GFAP and vimentin in a subclass of SCO-ependymocytes was demonstrated on differentially immunostained semithin sections. The present study confirms the glial nature of the SCO-ependyma, which has been a matter of debate recently. It appears from this investigation that the high degree of secretory activity in the SCO does not necessarily lead to the disappearance of glial IF proteins. Moreover, the SCO-cells belong to the expanding group of mature astroglia, which is characterized by coexpression of GFAP and vimentin. The morphological similarity between SCO-ependymocytes and tanycytes is underscored by their common immunoreactivity against these two IF proteins. In view of the absence of GFAP from the rat SCO, interspecific differences must be considered in the evaluation of the IF protein composition.     

乌梢蛇输精管道的组织学与免疫细胞化学观察

为探讨乌梢蛇(Zaocys dhumnades)输精管道结构与其功能之间的关系,该研究用一般光镜技术观察了乌梢蛇输精管道的显微结构及其年周期变化,并结合免疫细胞化学方法研究了雄激素受体(AR)、雌激素受体(ER)、孕激素受体(PR)和芳香化酶(Ar)在输精管和精巢中精子细胞表达的相关性.为验证该文在乌梢蛇输精管中观察到的大量精子和圆球状结构,用一般光镜技术还观察了黑眉锦蛇(Elaphe taeniura)、赤链蛇(Dinodon rufozonatum)与虎斑颈槽蛇(Rhabdophis tigrina lateralis)的输精管道.结果表明,乌梢蛇的输精管道主要由输出小管、附睾管与输精管构成;8-10月输出小管中有精子,8月—翌年1月附睾管中有精子,全年(除7月外)输精管中有大量精子;在输精管内首次观察到由多个精子细胞构成的圆球状结构,该结构与精巢中精子细胞的AR、ER、PR和Ar累计光密度值之间分别无显著差异.由于在乌梢蛇、黑眉锦蛇及赤链蛇的输精管内圆球状结构均可见精子细胞变态形成精子.因此,建议将蛇类输精管内圆球状结构命名为生精小球(seminiferous spherule).该文认为,蛇类精巢是精子形成的主要部位,而输精管内的生精小球是精子形成的另一个部位;附睾与输精管均可以储存精子,但输精管是精子储存的主要器官.     

The expression of inducible nitric oxide synthase (iNOS) in the testis and epididymis of rats with a dihydrotestosterone (DHT) deficiency

In our previous studies, we showed that a finasteride-induced DHT deficiency may cause changes in the morphology of the seminiferous epithelium without any morphological alteration of the epididymis. In this study, we demonstrated the constitutive immunoexpression of inducible nitric oxide synthase (iNOS) in the testis and epididymis of Wistar rats treated with finasteride for 28 days (the duration of two cycles of the seminiferous epithelium) and 56 days (the duration of one spermatogenesis). We noted that a 56-day finasteride treatment mainly caused a decrease in the level of circulating DHT, as well as a statistically insignificant decrease in the level of T. The hormone deficiency also led to a change in the iNOS immnoexpression in the testis and epididymis of the finasteride-treated rats. In vitro, DHT did not modify NO production by the epithelial cells of the caput epididymis even when stimulated with LPS and IFNγ, but it did give rise to an increase in NO production by the epithelial cells of the cauda epididymis without the stimulation. DHT did not have a statistically significant influence on estradiol production by cultured, LPS- and IFNγ-stimulated epithelial cells from the caput and cauda epididymis. In conclusion, our data clearly indicates that a finasterideinduced DHT deficiency intensifies the constitutive expression of iNOS in most rat testicular and epididymal cells, so it can be expected that the expression of inducible nitric oxide synthase (iNOS) could be regulated by DHT. On the other hand, the profile of the circulating DHT and T levels strongly suggests that the regulation of constitutive iNOS expression is complex and needs more detailed study.     

Localization of the epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in the bovine testis

In the last few decades, several growth factors were identified in the testis of various mammalian species. Growth factors are shown to promote cell proliferation, regulate tissue differentiation, and modulate organogenesis. In the present investigation we have studied the localization of EGF and EGFR in the adult bovine testis by means of immunohistochemical method. Our results demonstrated that EGF and EGFR were localized solely to the bovine testicular germ cells (spermatogonia, spermatocytes, and round spermatids). In contrast, the somatic testicular cells (i.e., Sertoli, Leydig, and myofibroblast cells) exhibited no staining affinity. EGF and EGFR were additionally detected in the epithelial lining of straight tubules and rete testis. Interestingly, the distribution of EGF and EGFR in the germ cells was mainly dependent upon the cycle of the seminiferous epithelium since their localization appeared to be preponderant during the spermatogonia proliferation and during the meiotic and spermiogenic processes. In conclusion, such findings may suggest that EGF and EGFR are important paracrine and/or autocrine regulators of spermatogenesis in bovine.     

Development of the islets, exocrine pancreas, and related ducts in the Nile tilapia, Oreochromis niloticus (Pisces: Cichlidae)

Pancreatic development and the relationship of the islets with the pancreatic, hepatic, and bile ducts were studied in the Nile tilapia, Oreochromis niloticus, from hatching to the onset of maturity at 7 months. The number of islets formed during development was counted, using either serial sections or dithizone staining of isolated islets. There was a general increase in islet number with both age and size. Tilapia housed in individual tanks grew more quickly and had more islets than siblings of the same age left in crowded conditions. The pancreas is a compact organ in early development, and at 1 day posthatch (dph) a single principal islet, positive for all hormones tested (insulin, SST-14, SST-28, glucagon, and PYY), is partially surrounded by exocrine pancreas. However, the exocrine pancreas becomes more disseminated in older fish, following blood vessels along the mesenteries and entering the liver to form a hepatopancreas. The epithelium of the pancreatic duct system from the intercalated ducts to the main duct entering the duodenum was positive for glucagon and SST-14 in 8 and 16 dph tilapia. Individual insulin-immunopositive cells were found in one specimen. At this early stage in development, therefore, the pancreatic duct epithelial cells appear to be pluripotent and may give rise to the small islets found near the pancreatic ducts in 16-37 dph tilapia. Glucagon, SST-14, and some PPY-positive enteroendocrine cells were present in the intestine of the 8 dph larva and in the first part of the intestine of the 16 dph juvenile. Glucagon and SST-14-positive inclusions were found in the apical cytoplasm of the mid-gut epithelium of the 16 dph tilapia. These hormones may have been absorbed from the gut lumen, since they are produced in both the pancreatic ducts and the enteroendocrine cells. At least three hepatic ducts join the cystic duct to form the bile duct, which runs alongside the pancreatic duct to the duodenum.     

Most genes encoding cytoplasmic intermediate filament (IF) proteins of the nematode Caenorhabditis elegans are required in late embryogenesis

Intestinal cells of C. elegans show an unexpectedly high complexity of cytoplasmic intermediate filament (IF) proteins. Of the 11 known IF genes six are coexpressed in the intestine, i.e. genes B2, C1, C2, D1, D2, and E1. Specific antibodies and GFP-promoter constructs show that genes B2, D1, D2, and E1 are exclusively expressed in intestinal cells. Using RNA interference (RNAi) by microinjection at 25 degrees C rather than at 20 degrees C we observe for the first time lethal phenotypes for C1 and D2. RNAi at 25 degrees C also shows that the known A1 phenotype occurs already in the late embryo after microinjection and is also observed by feeding which was not the case at 20 degrees C. Thus, RNAi at 25 degrees C may also be useful for the future analysis of other nematode genes. Finally, we show that triple RNAi at 20 degrees C is necessary for the combinations B2, D1, E1 and B2, D1, D2 to obtain a phenotype. Together with earlier results on genes A1, A2, A3, B1, and C2 RNAi phenotypes are now established for all 11IF genes except for the A4 gene. RNAi phenotypes except for A2 (early larval lethality) and C2 (adult phenotype) relate to the late embryo. We conclude that in C. elegans cytoplasmic IFs are required for tissue integrity including late embryonic stages. This is in strong contrast to the mouse, where ablation of IF genes apparently does not affect the embryo proper.