Developmental changes in Malpighian tubule cell structure.

Structural changes which occur in the Malpighian tubule yellow region primary cells during larval-pupal-adult development of the skipper butterfly Calpodes ethlius are described. The developmental changes in cell structure are correlated with functional changes in fluid transport (Ryerse, 1978a) in a way which supports osmotic gradient models of fluid secretion. Larval tubules are specialized for fluid secretion with deep basal infolds and elongate mitochondria-containing apical microvilli which provide channels in which osmotic gradients could be set up. The Malpighian tubule cells are extensively remodelled at pupation when fluid transport is switched off, but they persist intact through metamorphosis. At this time, the basement membrane doubles in thickness, the mitochondria are retracted from the microvilli and are isolated for degradation in autophagic vacuoles, and both apical and basal plasma membranes are internalized via coated vesicles for degradation in multivesicular bodies, which results in the shortening of the microville and the disappearance of the basal infolds. Mitochondria are re-inserted into the microvilli, and the basal infolds re-form in pharate adult stage Malpighian tubules when fluid secretion resumes. Adult tubules are similar in general structure to larval tubules and contain mitochondria in the microvilli and basal infolds. However, they differ from larval tubules in that they are capable of very rapid fluid transport, have a reduced tubule diameter and tubule wall thickness, a much thicker basement membrane and peripherally associated tracheoles. Mineral concretions of calcium phosphate accumulate in larval tubules, persist through metamorphosis and decline in number in adults, suggesting they serve some anabolic role.     

Rubidium reduces potassium permeability and fluid secretion in Malpighian tubules of Locusta migratoria, L

The basal membrane potential (V(b)) of Locusta Malpighian tubule cells in control saline results from its relatively high permeability to potassium. In the presence of 1 mM barium added to the control saline V(b) hyperpolarized from a mean resting potential of -72.1 mV to -90.1 mV. On substituting rubidium for potassium in the control saline, V(b) also hyperpolarized to a value of -91.4 mV. Rubidium was also similarly effective in hyperpolarizing the basal membrane even in the presence of control concentrations of potassium in the bathing medium. Substitution of rubidium for potassium also effected a approximately 50% reduction in the rate of fluid secretion. The action of inhibitors on V(b) in the presence of rubidium showed that V(b) under these conditions probably originated from the bafilomycin-sensitive electrogenic potential generated across the apical membrane by a V-type ATPase. The responses of V(b) to potassium, barium and rubidium and their inhibition of fluid secretion suggest the presence of a substantial rubidium-blockable potassium conductance located on the basal membrane of Locusta Malpighian tubule cells.     

Fine structure of the Malpighian tubules in the housefly, Musca domestica

The epithelium of the Malpighian tubules in the housefly is comprised of four distinct cellular types. Type I cells are characterized by the presence of intimate associations between infoldings of basal plasma membrane and mitochondria. On the luminal surface, cytoplasm is extended into microvilli which contain mitochondria. Membrane-bound vacuoles in the cytoplasm seem to progressively accumulate granular material. Type II cells have dilated canaliculi. Microvilli lack mitochondria. The Type III cell has not been reported previously in Malpighian tubules. It has very well-developed granular endoplasmic reticulum which contains intracisternal bundles of tubules. Cytoplasm contains numerous electron dense bodies. Type IV cells occur in the common duct region of the Malpighian tubules. Mitochondria do not extend into the microvilli.     

Transepithelial potential in Malpighian tubules of Rhodnius prolixus: lumen-negative voltages and the triphasic response to serotonin

Previous studies of the Malpighian tubules of Rhodnius reported lumen-negative values of transepithelial potential (TEP), and a characteristic triphasic change in TEP in response to stimulation of tubule fluid secretion by serotonin. TEP was measured using the Ramsay technique, in which electrodes are positioned in bathing and secreted fluid droplets for tubules isolated under paraffin oil. The validity of this method of TEP measurement has been questioned on the grounds that, in tubules of some species, it may permit shunting of current from lumen to bath through the cells or through the thin layer of fluid adherent to the surface of that portion of the tubule in the oil. The triphasic response of TEP to serotonin has been confirmed in this study of tubules of fifth instar Rhodnius prolixus using two different techniques that eliminate the possibility of shunting artefacts. From an initially negative value in unstimulated tubules ( approximately -25 mV, lumen-negative), TEP shifted to approximately -33 mV in phase 1, approximately +30 mV in phase 2 and approximately -32 mV in phase 3. TEP during each phase was similar irrespective of the measurement technique. Ion substitution experiments and the effects of specific pharmacological reagents support the proposal that the three phases of the response of TEP to serotonin correspond to sequential activation of an apical Cl(-) channel, an apical V-type H(+) ATPase and a basolateral Na(+):K(+):2Cl(-) cotransporter.     

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.     

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.     

Ultrastructural evidence for paracellular fluid flow in the Malpighian tubules of a larval mayfly

The ultrastructure of the Malpighian tubules of larvae of the Mayfly Ecdyonurus dispar (Ephemeroptera) is described. There are about 60 tubules, which consist of four distinct regions. The most proximal section (region I) appears to be responsible for fluid secretion. A unique feature is the presence of channels leading off the main lumen, which end close to the basal border of the cells. Microvilli are confined to these channels in region I. Region II is a short spiral region, the cells of which possess long basal folds and associated mitochondria. Region III is a simple conducting tube leading to one of six collecting ducts (region IV) arranged radially around the gut. In each collecting duct there are two cell types present. Type 2 cells are relatively simple, but give rise to numerous, long, microvilli-like projections. Type 1 cells possess long basal folds, and curious membrane whorls in the apical zone. Evidence is presented which suggest that water movements into region I takes place via the paracellular route. Region II is probably a reabsorptive region, but the function of region IV, based on ultrastructural evidence is more difficult to elucidate.     

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.     

The fine structure of the nephronic tubule of the mudskipper Periophthalmus koelreuteri (Pallas)

Ultrastructural examination of the head kidney of Periophthalmus koelreuteri (Pallas) (Teleostei, Gobiidae) revealed that the nephronic tubule cells are bound by tight junctions and desmosomes with little intercellular space. The first proximal segment (PI) consists of low columnar cells with well developed brush borders, indented nuclei, and numerous apical endocytic vesicles and lysosomes. A second cell type possessing clusters of apical cilia and lacking brush border and lysosomes is occasionally found between PI cells. The second proximal segment (PII) is formed of high columnar cells with brush border, regular spherical nuclei and numerous mitochondria located between well developed infoldings of the basal membrane. Single ciliary structures protrude into the lumen from PI and PII cells. The distal segment is lined by low columnar epithelium with few microvilli, regular spherical nuclei, numerous scattered mitochondria, and microbodies. The collecting tubule cells are cuboidal with few euchromatic nuclei, some mitochondria, and secondary lysosomes.     

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.     

Micromorphology of the malpighian tubules in the louse Pediculus humanus corporis (Anoplura)

The ultrastructure of the Malpighian tubes in human louse Pediculus humanus corporis has been studied. The cells of the Malpighian tubules have the uniform structure: the apical surface is covered with microvilli, the basal plasmatic membrana forms relatively small invaginations. The microvilli are most developed in cells of the proximal department of the Malpighian tubules. Microvilli of the apical surface of the cells do not contain mitochondria which are localized mainly in supranuclear part of the cell. Cells are lined with a homogenous basal membrane.     

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.     

Fine structure of the larval midgut of the fly Rhynchosciara and its physiological implications

The midgut of Rhynchosciara americana larvae consists of a cylindrical ventriculus from which protrudes two gastric caeca formed by polyhedral cells with microvilli covering their apical faces. The basal plasma membrane of these cells is infolded and displays associated mitochondria which are, nevertheless, more conspicuous in the apical cytoplasm. The anterior ventricular cells possess elaborate mitochondria-associated basal plasma membrane infoldings extending almost to the tips of the cells, and small microvilli disposed in the cell apexes. Distal posterior ventricular cells with long apical microvilli are grouped into major epithelial foldings forming multicellular crypts. In these cells the majority of the mitochondria are dispersed in the apical cytoplasm, minor amounts being associated with moderately-developed basal plasma membrane infoldings. The proximal posterior ventriculus represents a transition region between the anterior ventriculus and the distal posterior ventriculus. The resemblance between the gastric caeca and distal posterior ventricular cells is stressed by the finding that their microvilli preparations display similar alkaline phosphatase-specific activities. The results lend support to the proposal, based mainly on previous data on enzyme excretion rates, that the endo-ectoperitrophic circulation of digestive enzymes is a consequence of fluid fluxes caused by the transport of water into the first two thirds of midgut lumen, and its transference back to the haemolymph in the gastric caeca and in the distal posterior ventriculus.     

Ultrastructure of the Malpighian Tubule Cells in the Mosquito Larvae, Anopheles sinesis

Ultrastructure of epithelial cells constituting the Malpighian tubule of Anopheles sinesis last instar larvae was observed with electron microscope. Malpighian tubule consists of four long and narrow tubule structures with principal cells in typical absorptive cells and regenerative cells forming the simple epithelium. Apical plasma membrane of the principal cell is differentiated into microvilli with one mitochondrion in each microvilli. Basal plasma membrane had extreme infolding to form a canaliculi and a well developed mitochondria was attached in the infoldings. And, rER, ribosomes, and vacuoles were well developed inside the cells. However, there were two main cell types depending on the differentiation of cell organelles. Type 1 cell was cubic, forming the distal portion of Malpighian tubule. The length of microvilli was approximately 4 μm and the basal infoldings were introjected to the depth of 2 μm inside the cell. On the other hand, Type II cell that formed the main proxinal portion was a low squamous type cells with shorter 2 μm of microvilli and the basal infoldings were introjected to the depths of 4 μm inside the cell. As for vacuoles scattered inside the cells, they were regularly observed in both Type I and II and the Type II cells had better developed cellular organelles. Although regenerative cells were extremely small, their cellular organelles were developed and their overall electron density was high that they appeared darker than the principal cells.     

Choanocyte-like cells in the digestive system of the starfish Marthasterias glacialis (Echinodermata)

Two types of choanocyte-like cells have been found in the digestive tract of the starfish. Type I choanocytes are in the lining epithelium of all organs of the digestive system. These are narrow, columnar cells strongly anchored basally and expanded apically into a protuberance projecting into the lumen. A prominent flagellum surrounded by microvilli projects from the center of this protuberance. Apical cytoplasm contains numerous mitochondria, secondary lysosomes, and multivesicular bodies. A distinctive characteristic of these cells is a filament bundle that traverses the length of the cell from its region of attachment on the rootlet of the flagellar basal body to its terminus on the basal plasma membrane. Between the attenuated basal ends of type I cells are the nerve fibers of an intraepithelial nerve plexus. Thickness of the plexus is correlated with the quantity of type I cells in the epithelium. Type II choanocytes are in the cuboidal coelomic epithelium that forms the outer layer of digestive tract organs. These cells are smaller than those of type I, and they have an apical collar surmounted by a ring of 13 microvilli. Within the collar is a cup-shaped depression with a central flagellum. Coated vesicles, secondary lysosomes, and phagocytic infoldings are observed in and near the collar cytoplasm. Filament bundles similar to those in type I choanocytes are also observed in coelomic epithelial cells that are sufficiently tall. Injection of peroxidase into the stomach and ferritin into the coelom results in phagocytic uptake of these macromolecules by type I and type II choanocytes, respectively.     

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.     

An investigation into the effects of inhibitors of fluid production by Locusta Malpighian tubule Type I cells on their secretion and elemental composition

The intracellular elemental concentrations of K, Na, Cl, P, Mg and Ca within Type I cells of the Malpighian tubules of Locusta migratoria have been measured using electron probe X-ray microanalysis. The distribution of Na, K and Cl was not homogeneous within the cells and concentration gradients exist from basal to apical surfaces. The rate of secretion and the cationic composition of the secreted tubule fluid have also been determined. Furosemide (1 mM) inhibited fluid secretion by about 60%, raised the [Na(+)] but did not significantly alter the [K(+)] of the secreted tubule fluid. When Rb(+) replaced K(+) in the saline fluid secretion was also inhibited by about 60%, but no additional inhibition occurred by the simultaneous inclusion of furosemide. Thus, Rb(+) and furosemide probably act at the same transport site, and Rb(+) cannot substitute for K(+) at the basal membrane cotransporter. Bafilomycin (1 μM) dramatically inhibited fluid production by 85%, the [K(+)] of the secreted fluid was reduced by about 30% but the [Na(+)] was almost doubled. Furosemide, in common with other inhibitors of fluid secretion acting at the basal surface (ouabain and Rb(+)), caused a fall in intracellular [K] and a rise in [Na]. Bafilomycin, in common with N-ethyl maleimide, which acts at the apical surface, increased the intracellular [K] but did not affect the [Na].     

Invaginated apical vacuoles in the cells of the proximal convoluted tubule in the rat kidney

Summary Following perfusion fixation of the rat kidney with glutaraldehyde the proximal tubule cells display small apical vacuoles, large apical vacuoles, and apical vacuoles in which a part of the limiting membrane is invaginated into the vacuole. These invaginated apical vacuoles occur more frequently in proximal convoluted tubules than in proximal straight tubules. One tubular cell may contain apical vacuoles of different sizes and stages of invagination, ranging from larger vacuoles with a wide lumen and a small area of invaginated membrane to smaller elements with no apparent lumen and a large area of invaginated membrane. Invaginated apical vacuoles lie either singly in the cytoplasm or close to the membranes of other apical vacuoles, but never in contact with the cell membrane or the membranes of lysosomes, endoplasmic reticulum, Golgi apparatus, mitochondria and peroxisomes.These findings suggest that the invaginated apical vacuoles are not fixation artifacts, but rather develop in living state in cells of the proximal tubule from spherical endocytotic elements.Supported by the Deutsche Forschungsgemeinschaft (SFB 105)     

An ultrastructural study of Dirofilaria immitis infection in the Malpighian tubules of Anopheles quadrimaculatus

An ultrastructural study was conducted of the Malpighian tubules of Anopheles quadrimaculatus, both uninfected and following infection with Dirofilaria immitis. The Malpighian tubules in Anopheles are composed of primary and stellate cells. The primary cells are the predominant cell type and are characterized by the presence of membrane-bound, intracellular, mineralized concretions and large apical microvilli containing mitochondria. Following the infective blood meal, the microfilariae enter the primary cells of the Malpighian tubules and reside in the cytoplasm in a clear zone without a delimiting membrane. Cells in infected tubules differ from those in uninfected tubules in that the membranes of the vacuoles surrounding the concretions are disrupted in many specimens. The apical and basal cell membranes and the mitochondria associated with these are not disrupted during the first 6-8 days of infection. These observations differ sharply from those previously described in Aedes taeniorhynchus infected with D. immitis. The observations are consistent with the hypothesis that the extended transport capacity observed in previous physiological studies of An. quadrimaculatus infected with D. immitis are dependent on the prolonged normal ultrastructure of the apical microvilli, mitochondria, and basal membranes.     

Growth and differentiation of secondary malpighian tubules in the cockroach,Periplaneta americana

Four differentiated Malpighian tubules (primary tubules) extend from the junction of the midgut and hindgut in newly hatched Periplaneta americana. Secondary tubules begin to develop near the base of the primary tubules before hatching and successive nymphal molts. The newly initiated tubules undergo cell division and extensive elongation through the middle of the following intermolt period. During this time, the cells of the distal, middle, and lower middle tubule regions are surrounded by a cellular sheath, have few cytoplasmic processes extending along their basal surfaces, have a small or nonexistent lumen, and contain extremely dilated cisternae of endoplasmic reticulum. The cellular sheath differentiates into the muscle which coils around the mature tubule. Tubules which begin development toward the end of one intermolt period begin to undergo cytodifferentiation toward the end of the next intermolt period. By the middle of an additional intermolt period, the basal infoldings and microvilli of cells in the distal, middle, and lower middle regions have the conformations typical for those regions in differentiated tubules; granular concretions and stellate cells are present within the middle region of the tubule.