Ultrastructure of differentiated malpighian tubules from cockroach nymphs during the molting cycle

Differentiated Malpighian tubules of Periplaneta americana nymphs consist of four distinct regions. The distal, middle, and proximal regions are similar to the same regions in adult tubules. However, the transparent portion of the middle region was found to have ultrastructural characteristics different from those of the longer opaque segment of the middle region and the two other tubule regions. This newly distinguished region is called the lower middle region. Transitional zones, areas where cells show characteristics of two adjacent regions, are apparent between the distal and middle regions and between the middle and lower middle regions. The middle region of primary tubules undergoes an increase in autophagic activity and a modification of its basal infoldings and microvilli shortly before each molt. An increase in autophagic activity is also observed in the lower middle region near the time of molting.     

Ultrastructure of the tubular nephron of the lizard Podarcis (= Lacerta) Taurica

The proximal, intermediate, and distal convoluted tubules of the neprhon of Podarcis (= Lacerta) taurica were examined by electron microscopy. Proximal tubule cells have large, apical cytoplasmic protrusions and microvilli interpreted to function in urate secretion. Adjacent cells are bound apically by tight junctions and desmosomes but interdigitate in their basal region. This situation is repeated in the other tubules with significant differences in intercellular space width. The basal surfaces bear numerous cytoplasmic processes. The intermediate tubule has proximal and distal segments each with dark, ciliated, and light cells, the cuboidal dark cells with dense cytoplasm constituting the main bulk of the wall. As the cells of the proximal and distal segments resemble those of the proximal and distal convoluted tubules, respectively, the intermediate tubule is considered as a transition region. The ciliated cell body has two broad processes extending from the lumen, one to the basement membrane and one to a foot process of a light cell. The light cell is surrounded by dark and ciliated cells. It does not reach the lumen, but contacts the basement membrane through a process running below a ciliated cell to form a mushroom-shaped structure in tubule cross-section, the light cell process forming the stalk and a ciliated cell the cap. The cilia probably propel the glomerular filtrate towards the distal convoluted tubule. This latter tubule has initial, middle, and terminal zones, all nonciliated but with different lumen widths and cell shapes.     

Ultrastructure of the malpighian tubules of Schistocerca gregaria

The ultrastructure of the Malpighian tubules of the adult desert locust, Schistocerca gregaria, is described. Male and female adults possess about 233 tubules, which empty proximally into the midgut-ileal region of the alimentary canal by way of 12 ampullae. The tubules vary from 10 mm to 23 mm in length. About one third of them are directed anteriorly, attaching distally at the caeca, while the remainder are directed posteriorly, attaching to other tubules, the rectum or large tracheal trunks adjacent to the hindgut. The Malpighian tubules from all locations examined consist of three ultrastructurally distinct regions: proximal, middle, and distal, referring to their position relative to the midgut. All cell types possess ultrastructural features characteristic of ion transporting tissue, i.e., elaboration of the basal and apical membranes and a close association of these membranes with mitochondria. The distal and proximal segments are short (1.5-1.7 mm) and heavily tracheated, and each is composed of a single, distinct cell type. The middle region is the longest segment of the Malpighian tubule and is composed of two distinct cell types, primary and secondary. Both cell types are binucleate. The more numerous primary cells have large nuclei, contain laminate concretions in membrane-bound vacuoles, and possess large microvilli that contain mitochondria. The secondary cells are smaller and possess smaller nuclei. The microvilli are reduced and lack mitochondria. Secondary cells do not contain laminate concretions. The possible compartmentalization of ion and fluid transport function based on segmentation in the Malpighian tubules is discussed.     

Morphology and function of Malpighian tubules and associated structures in the cockroach, Periplaneta americana.

This paper describes the different regions of the Malpighian tubules and the associated structures (ampulla, midgut, ileum) in the cockroach, Periplaneta americana. There are about 150 tubules in each insect. Each tubule consists of at least three parts. The short distal region is thinner than the other parts and is highly contractile. The middle region comprises most of the tubule length and is composed of primary and stellate cells. Primary cells contain numerous refractile mineral concretions, while stellate cells have smaller nuclei, fewer organelles, simpler brush border, and numerous multivesicular bodies. Symbiont protozoa are sometimes present within the lumen of the middle region near where it opens into the proximal region of the tubule. The latter is a short region that drains the tubular fluid into one of the six ampullae. These are contractile diverticula of the intestine located at the midgut-hindgut junction. The ampulla is highly contractile, and consists of a layer of epithelial cells surrounding a cavity that opens into the gut via a narrow slit lined by cells of unusual morphology. The proximal region of the tubule and the ampulla resemble the midgut in that they have similar micromal origin and reabsorptive function for the proximal region of the tubule and for the ampulla. A number of inclusions found within the tubule cells are described, including peroxisomes and modified mitochondria. Current theories of fluid transport are evaluated with regard to physiological and morphological characteristics of Malpighian tubules. The possible role of long narrow channels such as those between microvilli and within basal folds is considered, as is the mechanism by which these structures are formed and maintained. Also discussed is the role of peroxisomes and symbionts in the excretory process.     

Modifications of the genital kidney proximal and distal tubules for sperm transport in Notophthalmus viridescens (Amphibia,Urodela, Salamandridae)

Male salamanders use nephrons from the genital kidney to transport sperm from the testicular lobules to the Wolffian duct. The microstructure of the epithelia of the genital kidney proximal tubule and distal tubule was studied over 1 year in a population of Notophthalmus viridescens from Crawford and Pike counties in central Missouri. Through ultrastructural analysis, we were able to support the hypothesis that the genital kidney nephrons are modified to aid in the transportation of sperm. A lack of folding of the basal plasma membrane, in both the genital kidney proximal and distal tubules when compared to the pelvic kidney proximal and distal tubules, reduces the surface area and thus likely decreases the efficiency of reabsorption in these nephron regions of the genital kidney. Ciliated epithelial cells are also present along the entire length of the genital kidney proximal tubule, but are lacking in the epithelium of the pelvic kidney proximal tubule. The exact function of these cilia remains unknown, but they may aid in mixing of seminal fluids or the transportation of immature sperm through the genital kidney nephrons. Ultrastructural analysis of proximal and distal tubules of the genital kidney revealed no seasonal variation in cellular activity and no mass production of seminal fluids throughout the reproductive cycle. Thus, we failed to support the hypothesis that the cellular activity of the epithelia lining the genital kidney nephrons is correlated to specific events in the reproductive cycle. The cytoplasmic contents and overall structure of the genital and pelvic kidney epithelial cells were similar to recent observations in Ambystoma maculatum, with the absence of abundant dense bodies apically in the epithelial cells lining the genital kidney distal tubule. J. Morphol. 275:914–922, 2014. © 2014 Wiley Periodicals, Inc.     

Maintaining the line of defense: regeneration of Cuvierian tubules in the sea cucumber Holothuria forskali (Echinodermata, Holothuroidea)

When irritated, individuals of the sea cucumber Holothuria forskali expel a few Cuvierian tubules which lengthen, instantly become sticky, and rapidly immobilize most organisms with which they come into contact. After expulsion, the lost tubules are readily regenerated. When only a few tubules have been expelled, there is often a latent period before the regeneration starts. In contrast, when many tubules have been expelled, the regenerative process starts immediately but proceeds in successive waves of 10 to 30 tubules that begin to regenerate at 10-day intervals. However, in all cases, the complete regeneration of a given tubule takes about 5 weeks and may be divided into three successive phases: an initial repair phase including the overall 48-h post-autotomy period, a true regenerative phase taking about 4 weeks to complete, and a growth phase of about one more week. Initial regeneration events occur by epimorphosis, cell proliferation being essential to the regenerative process, whereas late events occur mainly by morphallaxis, with migration of the newly differentiated cells. The mesothelium is the tissue layer in which cell proliferation is the most precocious and the most important, involving both peritoneocytes and undifferentiated cells (which seem to be dedifferentiated peritoneocytes). As regeneration proceeds, the percentage of undifferentiated cells regularly decreases in parallel with the differentiation of granular (adhesive-secreting) cells and myocytes. The myocytes then separate off from the mesothelium and migrate within the connective tissue layer. Three types of pseudopodial cells follow one another in the tubule connective tissue during regeneration. Type 1 cells have all the characteristics of echinoderm phagocytes and may have a fibroblastic function, cleaning the connective tissue compartment before new collagen synthesis starts. Type 2 cells are rather undifferentiated and divide actively. The presence of type 3 cells is closely associated with the appearance of collagen fibers, and it is suggested that they have a fibroblastic function. In the inner epithelium, cells also divide actively, but only those in which spherules have not yet differentiated in the basal intraconnective processes. It appears, therefore, that in the three tissue layers of the tubules, regeneration proceeds by cell dedifferentiation, then proliferation, and finally by differentiation. Cuvierian tubules thus constitute a very efficient defensive mechanism: their large number, sparing use, and particular regeneration dynamics make them an almost inexhaustible line of defense maintained at limited energy cost.     

Na-K]ATPase activity in proximal and distal tubules of the rat kidney: modification and application of a quantitative cytochemical technique

A modified cytochemical assay for [Na-K]ATPase in cryostat sections of kidney was further characterized and used to quantify activity in seven functionally distinct sites along the rat nephron. The activity of [Na-K]ATPase was defined as the difference in ATPase activity in specifically identified tubules contained in serial sections incubated with and without ouabain. Preincubation of sections with ouabain was required for maximal inhibition of [Na-K]ATPase activity in several distal sites. The concentration of ouabain necessary for maximal inhibition of activity was 3.0 mM and half-maximal inhibition was obtained in all regions with 30-100 microM ouabain. In distal sites, [Na-K]ATPase formed a higher proportion of total ATPase activity (60-80 per cent) than in proximal sites (20-40 per cent). Enzyme activity was quantified using two different methods. The first measured activity over the basal region of tubules and gave an index of the concentration of [Na-K]ATPase over the basal lateral infoldings of cells composing the tubule. The second read activity over the entire cross section of tubules and provided an estimate of [Na-K]ATPase per length of tubule. The highest activities over the basal basal region were obtained from tubules of the distal nephron including the inner (MALin) and outer (MALout) medullary ascending limb, distal convoluted tubule (DCT) and connecting segment (CS). Lower activities were obtained in proximal convoluted (PCT) tubules, proximal straight (PS) tubules and the papillary collecting duct (PD). Distal convoluted tubules contained the highest activity per length of tubule. Other sites contained lower levels of activity in the following order: MALin greater than MALout greater than PCT greater than PD greater than PS. The modifications introduced increase the sensitivity and precision of this assay and permit the application of this technique to studies of [Na-K]ATPase activity in the major functional regions of the rat nephron.     

Anatomy and fine structure of the alimentary canal of the spittlebug Lepyronia coleopterata (L.) (Hemiptera: Cercopoidea)

The alimentary canal of the spittlebug Lepyronia coleopterata (L.) differentiates into esophagus, filter chamber, midgut (conical segment, tubular midgut), and hindgut (ileum, rectum). The filter chamber is composed of the anterior extremity of the midgut, posterior extremity of the midgut, proximal Malpighian tubules, and proximal ileum; it is externally enveloped by a thin cellular sheath and thick muscle layers. The sac-like anterior extremity of the midgut is coiled around by the posterior extremity of the midgut and proximal Malpighian tubules. The tubular midgut is subdivided into an anterior tubular midgut, mid-midgut, posterior tubular midgut, and distal tubular midgut. Four Malpighian tubules run alongside the ileum, and each terminates in a rod closely attached to the rectum. Ultrastructurally, the esophagus is lined with a cuticle and enveloped by circular muscles; its cytoplasm contains virus-like fine granules of high electron-density. The anterior extremity of the midgut consists of two cellular types: (1) thin epithelia with well-developed and regularly arranged microvilli, and (2) large cuboidal cells with short and sparse microvilli. Cells of the posterior extremity of the midgut have regularly arranged microvilli and shallow basal infoldings devoid of mitochondria. Cells of the proximal Malpighian tubule possess concentric granules of different electron-density. The internal proximal ileum lined with a cuticle facing the lumen and contains secretory vesicles in its cytoplasm. Dense and long microvilli at the apical border of the conical segment cells are coated with abundant electron-dense fine granules. Cells of the anterior tubular midgut contain spherical secretory granules, oval secretory vesicles of different size, and autophagic vacuoles. Ferritin-like granules exist in the mid-midgut cells. The posterior tubular midgut consists of two cellular types: 1) cells with shallow and bulb-shaped basal infoldings containing numerous mitochondria, homocentric secretory granules, and fine electron-dense granules, and 2) cells with well-developed basal infoldings and regularly-arranged apical microvilli containing vesicles filled with fine granular materials. Cells of the distal tubular midgut are similar to those of the conical segment, but lack electron-dense fine granules coating the microvilli apex. Filamentous materials coat the microvilli of the conical segment, anterior and posterior extremities of the midgut, which are possibly the perimicrovillar membrane closely related to the nutrient absorption. The lumen of the hindgut is lined with a cuticle, beneath which are cells with poorly-developed infoldings possessing numerous mitochondria. Single-membraned or double-membraned microorganisms exist in the anterior and posterior extremities of the midgut, proximal Malpighian tubule and ileum; these are probably symbiotic.     

Electron-microscopic study of the excretory system of hungry females of the tick Hyalomma asiaticum P. Sch. et E. Schl. 2]

In H. asiaticum the cells of the Malpighian tubules and these of the rectal cas have the uniform structure: the apical surface is covered with microvilli, the basal plasmatic membrane forms relatively small invaginations. As to ultrastructural characters, there is no distinct division of the Malpighian tubule into departments. The distal ends of the tubules are not only somewhat enlarged and form the so-called ampulla cells of which are noticeably flattened. The microvilli and basal folds of the plasmatic membrane in this area of the tubule are indistinct. The cells of the ampulla and the neighbouring area of the tubule are characterized by the presence of inclusions with mucopolysaccharide secretion confined by the membrane. The microvilli are most developed on cells of the proximal ends of the Malpighian tubules. Well developed microvilli of the rectal sac form a striated border each containing a microtube inside. The basal invaginations are developed here better than in the cells of the Malpighian tubules.     

Excretion in the house cricket (Acheta domesticus): fine structure of the Malpighian tubules

An ultrastructural analysis of the Malpighian tubules of the cricket, Acheta domesticus, is presented. The excretory system of the cricket is unusual in that the 112 Malpighian tubules do not attach directly to the gut, but fuse to form a bladder-like ampulla which is joined to the colon by a muscular ureter. The tubules have three structurally distinct segments and consist of four cell types. Attached by basal lamina to the outer surface of the distal tip are nodules, consisting of small cells on membranous stalks. These are presumed to serve as attachments to the body wall. The distal 20% of the tubule is hyaline, consisting of a monolayer of squamous cells that appear to be secretory. The mid-tubule comprises 75% of the total length and is the primary region for fluid secretion. It is also characterized by having large numbers of laminate spheres in the cytoplasm of the cells. The proximal 5% of each tubule consists of more columnar cells and may function in fluid resorption. The relationship between structural features and known physiological functions are discussed.     

Analysis of nucleus activity in malpighian tubules of Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae) larvae by critical electrolyte concentration

Diatraea saccharalis (Fabricius) has cryptonephridial type Malphigian tubules (MT). This type of MT is characterized by the penetration of the distal part of the MT into the external walls of the rectum, which is usually lined with a perinephral membrane. The MT is divided into three differentiated regions: proximal, middle and distal. In this study, our objective was to compare the nuclear activities of each one of the three regions of the D. saccharalis MT by using a nuclear basophilic technique and critical electrolyte concentration with a toluidine blue stain at pH 4.0. This method allows differentiation of DNA/protein complexes in in situ and in vitro chromatin. MT chromatin structure in D. saccharalis is variable. Fifth instars have a more decondensed chromatin than fourth instars. The distal tubule region was the most decondensed region of the MT. Our data show an elevated genetic activity of the MT in the pre-metamorphosis period. The distal region of the MT has the highest observed activity, which may be associated with the re-absorption of useful components and the excretion of waste materials.     

白蜡虫马氏管的超微结构

白蜡虫Ericerus Pela的马氏管由两条黄色膨泡串状的端管和一条公共管构成,通过公共管与消化道相连。端管和公共管细胞结构相似,都具有非胶原质的基膜,高度发达的基褶, 长而致密的微绒毛,微绒毛无线粒体插入,细胞质中线粒体少,且随机分布。细胞质的绝大部分为两种矿质-尿酸颗粒结晶所占据,一种为不规则结晶,另一种为轮纹状结晶。白蜡虫马氏管可能发生了合胞化,其排泄方式可能是一种以滞留排泄为主,离子梯度排泄方式为辅的特有的排泄方式。     

Ricerche istochimiche e al micro scopio elettronico sui tubi malpighiani di Dacus oleae Gmel.

Summary The Dacus oleae larva possesses four Malpighian tubules, two anterior and two posterior ones, which in pairs enter into a ureter. Before opening into the gut, at the level of the transition zone between the mid- and hindgut, each ureter is dilated into an ampulla.The anterior tubules are divided into four regions: distal, transition, middle and proximal ones: while in the posterior tubules only middle and proximal segments are detectable. The distribution of the enzyme systems is indicated in Fig. 3, while the ultrastructural organization which is typical of the cells composing the different regions is schematically represented in Fig. 1. According to the ultrastructural and enzymatic findings, and the discussion on this subject in the literature, the authors are led to assume that in the distal segment occurs the segregation of uric acid, urates and calcium salts. In the transition segment, and still more in the intermediate one, an indiscriminate transport of water and solutes occurs from the haemocoel into the lumen of the tubule by pinocytosis. A fraction of the catabolites is precipitated as chromolipoidal pigments. The transition stages between cytosomes and pigment are described. Along with secretory phenomena the resorption of useful substances occurs in the proximal region. A similar function is performed by the ureter. In the ampulla, which is characterized by a conspicuous system of deep tubular infoldings both at the apical and basal surfaces of its cells, a massive water resorption is presumed to occur.     

The postnatal histology of the epididymis in buffalo (Bubalus bubalis).

The epididymis of buffalo is differentiated into 6 regions. The head represents the first 3 regions, the body the 4th region and the tail the 5th and 6th region. At 3-30 weeks, the epithelium is simple high columnar with few basal cells in region I, simple low columnar with few basal cells in region II, pseudostratified low columnar in regions III, IV, V, and pseudostratified low columar in region VI. As age advances, the epithelium increases in height and shows a tendency toward advanced pseudostratification. The basal cells are greater in number in regions III, IV and V than in the other regions of the epididymis. The epithelial cells contain stereocilia in region I at 3 weeks and regions III, IV and V at 30 weeks, whereas they are absent in regions II and VI. The tubules in region I are the smallest in diameter while the tubules in region VI have the largest diameter.     

Studies of vertebrate kidney. VI. Histochemical localization of phospholipids in kidneys of fishes from different habitats.

Six fish species living in different habitats such as fresh water, estuarine, and marine were studied for the distribution of phospholipids in various regions of the kidney. Well known histochemical methods, including those for choline-containing phospholipids were employed. The distribution pattern of phospholipids in different parts of the kidney such as the brush border, basal border and cytoplasmic granules of the proximal tubule cells, the cytoplasmic granules of the distal tubule cells, glomeruli and interstitial cells is given. Choline-containing phospholipids are present in the brush border of the proximal tubules of all the species studied and in the basal border of some. In the discussion an attempt at correlation of this histochemical pattern with the excretory functions and osmoregulation in these fishes is given.     

Structure of the kidney in the crab-eating frog, Rana cancrivora

The structure of the nephron in the ranid frog, Rana cancrivora, was studied by light and electron microscopy. This frog is the only amphibian species to live in mangrove swamps of very high salinity. The nephron consists of the following parts: renal corpuscle, ciliated neck segment, proximal tubule, ciliated intermediate segment, distal tubule, connecting tubule, and collecting duct. The distal tubule is located in the ventromedial region of the kidney, and the other tubules are situated in the dorsolateral region. Renal corpuscles are found between the two regions. Some renal corpuscles have a wide Bowman's space because of the small glomerulus within them. The proximal tubules are composed of columnar cells with a dense luminal brush border of long microvilli and numerous apical vesicles and vacuoles. The initial part of the distal tubule consists of heavily interdigitated cells, characterized by a very regular palisade arrangement of mitochondria. In the terminal part of the distal tubule, shorter mitochondria of the infolding cells are situated irregularly around the nucleus. The connecting tubule consists of principal cells and canaliculus cells. The collecting duct consists of columnar or cuboidal cells; cytoplasmic organelles are relatively sparse. The canaliculus cells are intercalated between principal cells from the terminal distal tubule to the proximal part of the collecting duct. Our findings indicate that the kidney of R. cancrivora is structurally similar to kidneys of other amphibians. These findings are discussed with regard to probable correlations between ultrastructure and function in R. cancrivora.     

Studies on vertebrate kidney. V. PAS-positive lipids in kidneys of fishes from different habitats.

Five fish species living in different habitats, i.e. fresh water, estuarine and marine, were studied for the distribution of PAS-positive materia in various regions of the kidney, 10 minutes' oxidation with 0-5 per cent. HIO4 being employed prior to treatment with Schiff's reagent. PAS-positive material was detected in different sites of the kidney, i.e. brush border of proximal tubules, proximal tubule cells' cytoplasm, distal tubule cells' cytoplasm, glomerulus, basal cell border of proximal tubules and the interstitial cells. Of these sites, the brush border of the proximal tubule of Scoliodon sorrakowah showed the presence of PAS-positive lipids. Elsewhere the PAS-positive reaction was due to carbohydrates. Free aldehyde groups were absent. In Tilapia mossambica and Labeo rohita, PAS staining was enhanced after chloroform-methanol extraction, particularly in the brush border of the proximal tubule. The significance of these findings is discussed.     

A structural basis for fluid secretion by malpighian tubules

The Malpighian tubules of Calliphora are described, emphasizing the possible role of surface specializations in solute-linked water transport. The tubules are composed of two cell types, primary and stellate, intermingling along the tubule length. The primary cells have long narrow basal infoldings and a microvillate luminal border, both intimately associated with mitochondria. The stellate cells have shorter and wider basal infoldings and their apical microvilli do not contain mitochondria. Application of the standing gradient hypothesis to this sytem provides a model for urine formation in which the local gradients for osmotic water flow occur within the long narrow channels of the basal infolds and microvilli of the primary cells. Stellate cells may modify the initial secretion by reabsorbing sodium.     

Morphology and ultrastructure of a secretory region enclosed by the venom reservoir in social wasps (Insecta,Hymenoptera)

The morphology and ultrastructure of the convoluted gland inside the venom reservoir of four species of social Vespidae are described. The cells of the venom gland (including the convoluted gland) can be divided into six groups: (1) epithelial cells, (2) glandular cells with the end apparatus secreting into the tubule inside the convoluted gland (internal or embedded tubule), (3) a continuous arrangement of glandular cells with the end apparatus secreting directly into the venom reservoir, (4) glandular cells that are loosely dispersed along the tubule lumen between the free tubules and the embedded tubule of the convoluted gland, (5) secretory cells of the free tubules and (6) duct cells. One kind of secretory cell, hitherto unknown and described in this paper (group 3), is characterized by the presence of a well-developed end apparatus, usually with enlarged extracellular spaces, but lacking the normally associated duct cells. The secretory cells contain several stacks of granular endoplasmic reticulum, but these are mainly concentrated in the middle of the cell. The basal half of the cells contains many lipid droplets. Although the function of the convoluted gland is not yet understood, an hypothesis is related to what is known of the function of reservoir secretory cells in solitary wasps. All wasp species studied showed the same organization of the convoluted gland, which clearly distinguishes their venom gland from that of Sphecidae.