Morphogenesis of intestinal villi. I. Scanning electron microscopy of the duodenal epithelium of the developing chick embryo

Development of villi in the duodenum of the chick was studied in stages ranging from 11 days of incubation to one week after hatching. Formation of definitive villi is preceded by development of a set of previllous ridges that run lengthwise along the duodenum. The first set of 16 previllous ridges (Set I) is complete by about 13 days of incubation; all ridges in the set are fairly uniform and proceed through their subsequent development in synchrony. Previllous ridges in Set I fold into a highly regular zigzag pattern between 14 and 16 days of incubation. Definitive villi develop from Set I ridges beginning at about 17 days when populations of distinct cells appear on the crests of the ridges between angles in the zigzag folds. Cells in these populations lack the rounded appearance of cells seen in earlier stages; their apical surfaces are densely covered with microvilli. A second set of villi (Set II) develops at about 16 days of incubation when about 16 rows of tongue-like flaps erupt between the previllous ridges of Set I. At hatching, Set II villi are still smaller than villi of Set I; this distinction disappears by about the fourth day after hatching. The significance of the morphological changes in epithelial cells is discussed in terms of several hypotheses bearing on the mechanisms of villus formation.     

The effects of thiourea and cortisone acetate on duodenal length and diameter and on morphogenesis of duodenal villi in chick embryos

Subnormal growth in length and diameter of the duodenum is encountered both in chick embryos injected with thiourea at 11 days of incubation, and in embryos injected with cortisone acetate at 14 days of incubation. Morphogenesis of previllous ridges and villi is retarded in thiourea-treated embryos. Morphogenesis of previllous ridges and villi in cortisone acetate-treated embryos at first seems to be slightly accelerated over that of controls, but the acceleration does not appear to be maintained, and the hormone may actually be inhibitory to normal morphogenesis of villi by 19 days of incubation. The results indicate that normal levels of endogenous thyroxine are necessary for normal growth in length and diameter of the duodenum and for normal morphogenesis of previllous ridges and villi. The possibility that adrenocorticoids may also play a role in the normal development of these parameters is indicated, although the evidence is inconclusive.     

On the development of the chick embryo colon: a computerized morphometric study

The quantitative aspects of the chick embryo colon organogenesis (proximal and distal tracts) have been studied from the 7th to the 15th day of incubation on histological sections by means of a computerized morphometric system. A semiautomatic digital system (Videoplan 2) was used. The area of total section, of the lumen, of the wall and its components (subserous stratum, muscle layer, lamina propria, epithelium) and the thickness of the epithelium and muscle layer were measured; the mean +/- S.E.M. of the obtained values was calculated. The percentage of shrinkage, due to histological procedures, was calculated for each day. The mean values for each day of incubation were modified on the basis of the shrinkage percentage. The differences between the mean values of the areas of the proximal and distal tract were statistically evaluated. Exponential curves and r coefficient were determined to evaluate the general growing pattern of the mean area of colonic wall components as a function of age. The main results demonstrate that in both tracts the component with the greatest growth was the muscle layer, while there are some differences between the proximal and distal tracts of the colonic anlage in the growing pattern of other wall components. The colonic anlage also shows a different developmental behaviour compared with that of the chick embryo ileum and duodenum.     

鹌鹑、红嘴鸥、黑翅长脚鹬胚胎期 小肠黏膜表面形态结构

应用扫描电镜观察了鹌鹑（Coturnix coturnix）、红嘴鸥（Larus ridibundus）、黑翅长脚鹬（Himantopus himantopus）胚胎期小肠黏膜的形态变化。鹌鹑与红嘴鸥胚胎发育过程中,小肠黏膜形态结构可分为3个阶段。第一阶段为鹌鹑卵孵育第10 ~ 11天、红嘴鸥卵孵育第13 ~ 14天（相当于胚胎发育60%的阶段）,小肠黏膜为山脊状纵行皱襞;第二阶段为鹌鹑卵孵育第12 ~ 13天、红嘴鸥卵孵育第15 ~ 16天（相当于胚胎发育70%的阶段）,小肠黏膜为“W”形板状皱襞;第三阶段为鹌鹑卵孵育第14 ~ 17天、红嘴鸥卵孵育第17 ~ 22天（相当于胚胎发育到80% ~ 100%阶段）,小肠黏膜为指状绒毛。黑翅长脚鹬卵孵育第10天（相当于胚胎发育到60%的阶段）,小肠黏膜为山脊状纵行皱襞;从卵孵育第12天（相当于胚胎发育到70%的阶段）一直到孵出小肠黏膜均为“W”形板状皱襞。初步判断3种鸟小肠黏膜形态发生这种有规律的变化可能是鸟类对其祖先系统发育的重演。     

A UNIQUE JUNCTIONAL STRUCTURE FOUND IN BLASTULA CELLS OF THE NEWT, TRITURUS PYRRHOGASTER

The outermost cell layer of the animal half of the newt blastula ( Triturus pyrrhogaster ) was examined to investigate intercellular junctions by transmission and scanning electron microscopy. A unique structure is observed at the terminal region of the intercellular junction. The structures are cytoplasmic ridges elevated from the cell surfaces, and their inner part is filled with spaces of various sizes. It is supposed that these ridges result from the encounter of cytoplasmic folds protruding from two neighboring cells.
Below the ridges, there is a short close junctional area which is followed by a long region of intercellular space intermittently bridged by cytoplasmic projections. Microvillus-like cytoplasmic processes on the apical cell surfaces, and microfilaments and microtubules in subsurface regions are observed in this material as in many other embryonic cells of amphibians.     

Distribution of Intercellular Material in the Apical Part of Pea Seedlings

The sections from the upper part of the third internode, counted from the seed, in etiolated pea seedlings are studied for the distribution of two types of Intercellular spaces: the transparent and the dark. The transparent spaces represent the result of the water logging of passages originally water-lined, while the dark spaces are lined with a lipid-containing substance which may be impregnated with melted paraffin and which forms a ramifying network within the tissue. The intercellular material of the dark spaces has the appearance of a tube, since it concentrically lines the intercellular space, leaving a lumen for air or gas. It is a plastic, isotropic subslance which may be cut transversely and identified in successive sections of the inlernode as belonging to the same continuous material from the intercellular space. The dark and the transparent spaces have a distinct pattern of distribution in the growing internode, and the relative quantity of dark spaces present at different levels of the internode may be measured. The dark spaces predominate in those parts of the stem with a greater potential for growth, while the transparent spaces are located in the regions where growth has ceased or subsided. Since the paraffin is infiltrated instantaneously throughout the network of dark intercellular spaces, it is possible that these may represent a channel for the translocation of substances.     

Occluding-like junctions at mesaxons of central myelin in Anolis carolinensis are not 'tight'. A freeze-fracture-protein tracer analysis.

The junctional complexes of the myelin sheath of central nervous system axons in the American chameleon, Anolis carolinensis, exhibit an intramembrane ridge and groove construction in freeze-fracture replicas that has usually been interpreted in other organisms as evidence for an occluding or tight intercellular junction. Close examination of PF fracture face ridges, however, shows them to be made up of discontinuous rows of particles of variable length separated by frequent gaps of non-uniform width. Introduction of horseradish peroxidase into the intercellular milieu of the lizard central nervous system is followed by appearance of this protein in interlamellar spaces of the myelin sheath and in the intercellular spaces containing focal membrane fusions that correspond precisely in position and center-to-center spacing to the ridges and grooves in platinum replicas of the same tissue. Since the junctional ridges on PF fracture faces in these mesaxonal junctional complexes are conspicuously discontinuous and since the areas within the myelin sheath where these junctional complexes are located inner and outer mesaxons) are readily permeated by exogenous protein tracer, it is concluded that the junctional complexes of central myelin mesaxons, heretofore incorrectly interpreted as functionally tight, are actually very leaky and probably contribute only to the structural stability of the myelin sheath architecture.     

Regional differences in hindgut structure and function in the nutria,Myocastor coypus

Summary Morphologically the large intestine of the nutria resembles that of other caviomorphs, notably the guinea pig. The cecum is voluminous: it contributes 8.6% of the total intestinal length and 12.7% of the total intestinal surface area (considering the surface enlargement factor). It contains 27–32% of the wet ingesta and 20–23% of the dry matter in the gastrointestinal tract. In the colon the corresponding figures are: 21.8% of length, 12.6% of surface area, 16–21% of wet ingesta, and 16–40% of dry matter. The colon can be subdivided both structurally and functionally into two sections, the proximal and the distal colon, the border between the two being the apical flexure of a long parallel loop. The proximal colon (42% of colonic length) displays on the mucosal surface of its mesenterial side a narrow furrow bordered by ridges, which is absent in the distal colon. The ridges contain subepithelial accumulations of coiled tubuloalveolar mucoid glands, entwined by bundles of muscle fibers. Determinations of nitrogen in the contents near the furrow suggest a concentration of bacteria in this part of the lumen. It is hypothesized that the structural differentiations of the proximal colon provide mechanisms for the transport of bacteria from the proximal colon back into the cecum to maintain the fermentation function. The slopes of the longitudinal profiles for dry matter and for concentrations of sodium, potassium and calcium in the luminal contents change at the tip of the parallel loop. The electrical potential difference intestinal lumen — blood is particularly large in the proximal colon, indicating active electrogenic ion transport in this region. SEF surface enlargement factor - CSM colonic separation mechanism - PEG polyethylene glycol - DM dry matter Dedicated to Prof. K.-E. Wohlfarth-Bottermann, Institut für Cytologie und Mikromorphologie der Universität Bonn, Bonn, Federal Republic of Germany, on the occasion of his 65th birthday     

The development of the neurons of the glossopharyngeal (IX) and vagal (X) sensory ganglia in chick embryos

The timetable of cell generation, neuronal death and neuron numbers in the fused proximal glossopharyngeal (IX) and vagal (X) ganglion and distal IX and X ganglia were studied in normal and nerve growth factor (NGF) treated chick embryos. 3H-thymidine was injected between the 3rd and 7th days of incubation and embryos sacrificed on the 11th day. Neurons in the distal IX and X ganglia were generated between the 2nd and 5th days of incubation, the peak mitotic activity occurring on the 4th and 3rd days, respectively. Neurons of the proximal IX and X ganglion were generated between the 4th and 7th days, with maximum neuron generation on the 5th day of incubation. Counts of neurons in the 3 ganglia between the 5th and 18th days of incubation showed a maximum of 22,000 on the 8th day in the proximal IX and X ganglion and this decreased to 12,000 by the 13th day. In the distal IX ganglion, the neuron number decreased by 44% from 4,500 on the 6th day to 2,500 by the 11th day. A similar decrease of 43% was found in the distal X ganglion, the neuron number falling from 11,500 on the 7th day to 6,500 by the 11th day of incubation. Neuronal cell death in these ganglia extended from the 5th to the 12th day of incubation, maximum cell death occurring at or after the cessation of mitotic activity. NGF administration from the 5th to the 11th day of incubation did not have a measurable effect on the neurons of proximal IX and X and distal IX ganglia, but increased neuronal survival by 30% in the distal X ganglion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Glycoconjugates of the cecal intestinal epithelium of the chick before and after hatching: histochemical study performed with peroxidase-conjugated lectins]

A battery of seven different horseradish peroxidase-labelled lectins (PNA, ConA, DBA, SBA, LTA, WGA and UEA I) was used to study the distribution and changes of carbohydrate moieties of glycoconjugates in the caecal epithelium (proximal and distal tracts) of the chick embryo and of the 3 days old chicken. The chief results showed that: 1. The appearance of some sugar residues was earlier observed at the epithelium of the distal tract than the proximal one (Tab. 2). 2. The presence of sialic acid was detected only after hatching (Fig. 4, Tab. 2). 3. During the embryonic caecal development enterocytes and goblet cells were characterized by the presence of the same sugar residues (Tab. 2). 4. By a quantitative point of view, differences in sugar residues content between the epithelium of the proximal and distal tract were observed. The epithelial cells of the distal tract were generally characterized by an higher content of saccharide moieties (Tab. 2). 5. At the end of the incubation period and after hatching enterocytes and goblet cells showed differences in content of some sugar residues (Fig. 1-3, Fig. 5-8, Tab. 2).     

Circadian range of entrainment in the semilunar eclosion rhythm of the marine insect clunio marinus

The semilunar eclosion of the intertidal chironomid Clunio is controlled by a semilunar timing of pupation in combination with a daily timing of emergence. This results in reproductive activities of a laboratory population every 15 days at a distinct time of day (in nature mostly in correlation with the afternoon low water time on days with spring tides). The entrainment of the timing processes has been tested under various periods of the daily light-dark cycle in order to check the circadian organization of the timing mechanisms as suggested for the perception of the semilunar zeitgeber situation (a distinct phase relationship between the 24 h light-dark cycle and the 12.4 h tidal cycle recurring after every 15th light-dark cycle, named semimonthly zeitgeber cycle) as well as for the daily zeitgeber (the 24 h light-dark cycle). With respect to the semilunar timing, a strong entrainment was only possible in semimonthly zeitgeber cycles with light-dark cycle periods close to the 24-h day (light-dark cycles of 10:10 to 14:14). This limited circadian range of entrainment of an endogenous circasemilunar long-term rhythm (syn. oscillator) conforms with the hypothesis for a circadian clock component as an intrinsic part of the semilunar zeitgeber perception.The range of entrainment for the daily timing was obviously wider which may be discussed either in relation to a multioscillatory circadian organization of the midges or in relation to different coupling characteristics of one circadian oscillator during semilunar and daily timing.     

Relationship between the cartilage canal and the perichondrium in the rat proximal tibial epiphysis

One of the most widespread hypotheses for chondral canal morphogenesis suggests that the canal is an extension of the perichondrium. To study the possible relation between perichondrium and chondral canal morphology, the proximal epiphyses in the tibias of 42 rats were studied from birth to their 29th day. The study was divided into three periods: from birth to the 4th day before canal appearance; from the 5th day, the moment of canal appearance, until the appearance of the secondary ossification center of the epiphysis on the 9th day; the 3rd ran from this point on the 10th day until its full development. We have also divided the canal into three regions: entrance, neck and bottom. The central portion (lumen) and canal wall were analyzed in each region. Our results show the perichondrium to be a complex structure, composed of a series of cellular layers in a biphasic extracellular matrix (eosinophil and basophil). The canal walls are lined by a layer of elongated cells. In the lumen there are many different cell types: fibroblasts, histiocytes, multinuclear giant cells and multivacuolated cells. Our study of the canal, its walls and lumen show no morphological structure that is reminiscent of the perichondrium. These results suggest that the canal is not itself a continuation of the perichondrium.     

The early effect of dextran sodium sulfate administration on carbachol-induced short-circuit current in distal and proximal colon during colitis development

Increased colonic Cl(-) secretion was supposed to be a causative factor of diarrhea in inflammatory bowel diseases. Surprisingly, hyporesponsiveness to Cl(-) secretagogues was later described in inflamed colon. Our aim was to evaluate changes in secretory responses to cholinergic agonist carbachol in distal and proximal colon during colitis development, regarding secretory activity of enteric nervous system (ENS) and prostaglandins. Increased responsiveness to carbachol was observed in both distal and proximal colon after 3 days of 2 % dextran sodium sulfate (DSS) administration. It was measured in the presence of mucosal Ba(2+) to emphasize Cl(-) secretion. The described increase was abolished by combined inhibitory effect of tetrodotoxin (TTX) and indomethacin. Indomethacin also significantly reduced TTX-sensitive current. On the 7th day of colitis development responsiveness to carbachol decreased in distal colon (compared to untreated mice), but did not change in proximal colon. TTX-sensitive current did not change during colitis development, but indomethacin-sensitive current was significantly increased the 7th day. Decreased and deformed current responses to serosal Ba(2+) were observed during colitis induction, but only in proximal colon. We conclude that besides inhibitory effect of DSS on distal colon responsiveness, there is an early stimulatory effect that manifests in both distal and proximal colon.     

Induction of feather malformations in chick embryos by cadmium: protection by zinc

Various doses of cadmium chloride were injected to chick embryos between the seventh and 14th day of incubation. Doses over 15 micrograms/egg produced high mortality and, when injected between the tenth and 11th day, widespread curling of the feathers in the surviving embryos. A different type of malformation, consisting of hemorrhagic atrophy of the distal part of the feathers, was observed in the embryos injected with similar doses during the 12th day. No feather malformations were observed in embryos injected before the ninth or after the 12th day of incubation. The simultaneous injection of an equimolar amount of zinc sulfate prevented the feather malformations.     

Funktionsstrukturen im Oocytenkern von Locusta migratoria

Examination of living oocyte nuclei of Locusta migratoria has revealed the presence of thread-like struktures. They are paired and are thought to be the uncoiled chromosomes since they are broken into fragments by treatment with DNase. The greater part of the threads carries lateral loops like the lampbrush chromosomes of Amphibia (Fig. 14). A smaller part has no loops hut bears a series of conspicious granules with bright appearance under positive phase contrast optics (pearl-string segments) (Fig. 2). — The visibility of the chromosomes has been investigated in solutions with several ions. In hypertonic media the chromosomes contract, the granules fuse, and the pearl-string segments become lumpy (Fig. 21). In nitrogenous atmosphere and if kept at low temperature the pearl-string structures are likewise transformed into a few lumps (Fig. 19). After return to normal conditions they reconstitute their characteristic beaded appearance. — In autoradiographs obtained by injection of H³-uridine into the body cavity and by incubation of isolated nuclei in vitro, a rather uniformly distributed labelling occurs over the oocyte nuclei up to 30 min incubation time (Fig. 23). With prolonged incubation the activity of the pearl-string segments becomes more intense than the labelling of the lampbrush chromosomes (Fig. 24). After treatment with actinomycin RNA synthesis is stopped, the pearl-string axes and the lampbrush chromosomes contract, and the granules disappear more and more (Figs. 25-28). — The pearlstring segments look very much like the nucleoli in the oocytes of Amphibia, where the nucleolar substance is likewise distributed as a series of beads of rather uniform size on an axis. Therefore, the pearl-string structures may have nucleolar function in Locusta too. If so, the only difference to the Amphibian nucleoli would be the continued attachment to the lampbrush chromosomes.     

Regulation of goblet cell differentiation by calcium in embryonic chick intestine

The potential role of calcium as a regulator of goblet cell differentiation was tested by culturing duodenal explants from 14-day chicken embryos in media containing a range of calcium concentrations. Extracellular calcium in vitro approximated the range of plasma calcium concentration that was found to rise by 16% during the third week of embryonic development. As ionic calcium was increased from 0.9 to 2.0 mM in 48-h cultures, the number of goblet cells per 100 ridges increased progressively from 29 +/- 2.2 to 158 +/- 6.9 while attaining a distribution on the previllous ridges similar to that in ovo. The rate of goblet cell differentiation in vitro exceeded that in ovo when extracellular calcium was increased to 1.1 mM and was maximal at 1.6-2.0 mM calcium. Neither the growth of previllous ridges nor gross morphology of the tissue was altered as extracellular calcium was increased. Goblet cell differentiation in 1.3 mM calcium was stimulated by 0.1 microM calcium ionophore and inhibited by the calmodulin antagonist trifluoperazine, indicating an intracellular site for calcium action. These results indicate that calcium plays a primary role in regulating goblet cell differentiation in embryonic intestine and suggest that rising levels of plasma calcium influence the rate of differentiation in ovo.     

PATHS OF TRANSTUBULAR WATER FLOW IN ISOLATED RENAL COLLECTING TUBULES

The cells of perfused rabbit collecting tubules swell and the intercellular spaces widen during osmotic flow of water from lumen to bath induced by antidiuretic hormone (ADH). Ouabain had no influence on these changes. In the absence of net water flow intercellular width was unaffected when tubules were swollen in hypotonic external media. Therefore, during ADH-induced flow widening of intercellular spaces is not a consequence of osmotic swelling of a closed intercellular compartment containing trapped solutes, but rather is due to flow of solution through the channel. Direct evidence of intercellular flow was obtained. Nonperfused tubules swollen in hypotonic media were reimmersed in isotonic solution with resultant entry of water into intercellular spaces. The widened spaces gradually collapsed completely. Spaces enlarged in this manner could be emptied more rapidly by increasing the transtubular hydrostatic pressure difference. In electron micrographs a path of exit of sufficient width to accommodate the observed rate of fluid flow was seen at the base of the intercellular channel. It is concluded that the intercellular spaces communicate with the external extracellular fluid and that water, having entered the cells across the luminal plasma membrane in response in ADH, leaves the cells by osmosis across both the lateral and basilar surface membranes.     

Luminally released serotonin stimulates colonic motility and accelerates colonic transit in rats

Enterochromaffin (EC) cells of the epithelial cells release 5-HT into the lumen, as well as basolateral border. However, the physiological role of released 5-HT into the lumen is poorly understood. Concentrations of 5-HT in the colonic mucosa, colonic lumen, and feces were measured by HPLC in rats. To investigate whether intraluminal 5-HT accelerates colonic transit, 5-HT and (51)Cr were administered into the lumen of the proximal colon, and colonic transit was measured. To investigate whether 5-HT is released into the lumen, we used an ex vivo model of isolated vascularly and luminally perfused rat proximal colon. To investigate whether luminal 5-HT is involved in regulating stress-induced colonic motility, the distal colonic motility was recorded under the stress loading, and a 5-HT(3) receptor antagonist (ondansetron, 10(-6) M, 0.5 ml) was administered intraluminally of the distal colon. Tissue content of 5-HT in the proximal colon (15.2 +/- 4.3 ng/mg wet tissue) was significantly higher than that in the distal colon (3.3 +/- 0.7 ng/mg wet tissue), while fecal content and luminal concentration of 5-HT was almost the same between the proximal and distal colon. Luminal administration of 5-HT (10(-6)-10(-5) M) significantly accelerated colonic transit. Elevation of intraluminal pressure by 10 cmH(2)O significantly increased the luminal concentration of 5-HT but not the vascular concentration of 5-HT. Stress-induced stimulation of the distal colonic motility was significantly attenuated by the luminal administration of ondansetron. These results suggest that luminally released 5-HT from EC cells plays an important role in regulating colonic motility in rats.     

Localization of Pathogenesis-Related Proteins in the Epidermis and Intercellular Spaces of Tobacco Leaves after Their Induction by Potassium Salicylate or Tobacco Mosaic Virus Infection

The localization of pathogenesis-related (PR) proteins inducedin tobacco leaves by treatment with potassium salicylate ora hypersensitive response to tobacco mosaic virus (TMV) infectionwas studied using immunochemical methods. Total PR protein levelsincreased with time after these treatments. The proportion ofPR proteins in the intercellular spaces to the total contentin the leaf discs rapidly rose in the later stage of the treatmentto about 75% on the 9th day after salicylate treatment and tomore than 80% on the 6th to 9th day after TMV inoculation. After5 days of salicylate treatment, the amounts of PR proteins inthe peeled leaf epidermis were two fold those in the mesophylltissue. Only five percent or less of the total PR proteins inthe epidermal and mesophyll tissues of salicylate-treated leaveswere detected in the isolated epidermal and mesophyll protoplasts.The sugar content in highly purified PR la, lb and lc was lessthan one mole of monosaccharide per mole of each protein. Theseresults show that the PR proteins are non-glycoproteins secretedinto the intercellular spaces. (Received January 16, 1987; Accepted July 14, 1987)