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Influence of the mesonephros on the development of fetal mouse ovaries following transplantation into adult male and female mice.

We previously reported that fetal mouse ovaries frequently develop testicular structure following transplantation into adult male mice. The mechanism involved in gonadal sex reversal of ovarian grafts is not known. In the present study, we examined the influence of the adjacent mesonephros on development of the ovarian grafts. The results show that (1) when fetal ovaries were transplanted with the attached mesonephros, the frequency of ovotestis development was higher in male hosts than in female hosts, (2) the fetal ovaries that had been separated from mesonephros developed testicular structures more frequently than those with the mesonephros, and the incidence of ovotestis development was comparable in male and female hosts, (3) removal of the cranial or caudal half of the mesonephros resulted in a similar frequency of ovotestis development, and (4) when fetal ovaries were separated and reattached to the mesonephros, they developed testicular structures at a frequency similar to that of ovaries left attached to the mesonephros, and the sex of mesonephroi reattached to ovarian grafts did not influence the incidence of ovotestis development. These findings suggest that fetal ovaries can develop testicular structures after transplantation regardless of the sex of host, and that the adjacent mesonephros protects ovarian grafts from masculinizing stimuli more efficiently in female host than male hosts.

Animals

A growth-promoting influence from the mesonephros during limb outgrowth.

It has been suggested that the mesonephros has a role in normal limb development. This hypothesis was directly tested by removing the mesonephros adjacent to the presumptive limb region of stage 12-18 chick embryos using microsurgery or laser ablation. The experimental manipulation resulted in reduced limb outgrowth on the operated side. The poor limb outgrowth was correlated with either the lack of or the presence of a rudimentary mesonephros on the operated side. Furthermore, the presence of nephric tissue in limb bud organ culture enhanced growth and morphological differentiation of cartilage formed in culture. In vivo, the influence of the mesonephros resulted in significantly higher cell proliferation in the adjoining medial half of the limb mesoderm compared with the lateral half. The removal of the mesonephros adjoining the prospective limb region reduced the number of dividing cells in the medial mesoderm. The higher proliferation in the medial limb mesoderm is significant to limb outgrowth since grafting experiments showed that most of the cells that form the limb are derived from the medial mesoderm. The results suggest that the influence from the mesonephros may provide some signal for limb outgrowth.

Animals

The chicken embryonic mesonephros synthesizes link protein, an extracellular matrix molecule usually found in cartilage.

Link protein is a macromolecule that is relatively abundant in the extracellular matrix of cartilage, where it acts as a stabilizing component in aggregates of the large chondroitin sulfate proteoglycan and hyaluronic acid. In the present study, link protein transcripts were demonstrated in the chicken embryonic mesonephros by RNA in situ hybridization using a cartilage link protein cDNA probe. The link protein transcripts of the mesonephros are of the same size as those seen in cartilage. In addition, mesonephroi contain a protein that is immunologically reactive with a link protein polyclonal antiserum and this protein is identical in size to link protein isolated from cartilage. No transcripts for cartilage proteoglycan core protein were detected in the mesonephros. Type II collagen and cartilage matrix protein transcripts were also not detectable in the mesonephros. From previous data on chondrogenesis in the developing limb bud, the transcription of link protein and the proteoglycan core protein genes appeared to be spatially and temporally regulated in a coordinated fashion. However, the presence of link protein transcripts in the mesonephros, independent of cartilage proteoglycan core protein gene expression, indicates that these genes can be regulated independently of each other.

Animals

Changes in the protein spectrum of the chick embryo mesonephros and metanephros in the course of development.

The protein spectra of two fractions (the soluble and the membrane fraction) of chick embryo kidney homogenates were isolated by electrophoresis on polyacrylamide gels with the aim of detecting the kidney differentiation process at the molecular level and, at the same time, of evaluating similarities in the construction of the mesonephros and metanephros at this level. Corresponding stages of the above two types of kidney were chosen for studying changes in protein structure during differentiation--i.e. the outset of differentiation (the 6-day mesonephros, the 11-day metanephros) and the stage of full maturity (the 14-day mesonephros, the 20-day metanephros). A total of 36 proteins was distinguished. The analysis of the protein spectra showed that the number of proteins changes but slightly during differentiation; the protein composition of the two types of kidney during differentiation altered by 20-35% of the total number of proteins; the similarity of the protein composition of the corresponding stages of mesonephros and metanephros, expressed as the proportion of the number of identical proteins, was greater than the mutual similarity of different developmental stages of the same type of kidney. The percentage of different proteins at corresponding stages of the kidneys varied from 5% to 23% of the total number of proteins detected.

Animals

Gonadal development of the chick embryo following microsurgically caused agenesis of the mesonephros and using interspecific quail-chick chimaeras.

Mesonephric agenesis was achieved by microsurgical excision of the left Wolffian duct and the underlying intermediate mesoderm of different regions between somites 16 and 23 in chickens after 50-52 h of incubation (stage 14 HH). Quail-chick chimaeras were produced by transplantation of corresponding quail tissue in the region of somites 18-21. A morphometrical analysis of the mesonephric and gonadal area in cross sections shows that the intermediate mesoderm from somites 16 to 23 develops into the mesonephros. A partial agenesis of the mesonephros brought about by removal of the intermediate mesoderm at the level of somites 18 to 21 at stage 14 leads to a mean reduction of the gonadal volume of 37.8% compared to the volume of the untreated side at stage 30. Transplantation of quail intermediate mesoderm in this region of the excision results in development of a hybrid mesonephros. Consequently, the gonads are invaded and colonized by quail cells mobilized from mesonephric corpuscles examined at stage 30, 35 and 36. These results are discussed in terms of the origin of the gonadal stroma during this developmental period; they show that in the region from the third to the sixth segment the ventromedial part of the differentiating mesonephros participates in the contribution of stromal cells to the gonad.

Animals

Morphological observations on the mesonephros in the postnatal opossum, Didelphis virginiana.

The mesonephros of the opossum persists for 3-4 weeks into the postnatal period. Based on our observations of its structure, and the vital dye experiments of others, it appears that the opossum mesonephros is functional during the first 10 days of the postnatal period. The mesonephros of the newborn opossum consists of 35-45 nephron units which are structurally very similar to metanephric nephrons except that they lack a loop of Henle. By the end of the first postnatal week, regression which proceeds in a craniocaudal direction, is observed. By the end of the second week most nephrons show some signs of regression. The regressing mesonephric nephrons are replaced by connective tissue.

Animals

The problem of sampling homologous groups of nephrons during development of the chick mesonephros.

With the aim of defining homologous groups of nephrons in the chick mesonephros at different stages of development, we opened the abdominal wall of 295 White Leghorn embryos at stage HH 26 and marked off 6 segments of equal length at different levels of the organ by Seichert's plastic linear marking method (Seichert 1965), using marks previously introduced into the vertebral column at the same levels as a guide. The embryos were removed after 2-6 days, weighed and classified according to weight. The primordia of the marked vertebrae were identified. The length of the artificially defined segments or their relationships to both poles of the mesonephros were evaluated in projections. The portion of the mesonephros above Th 3 level shorten during the period of observation, while the more caudal segments grow in a craniocaudal gradient. Homologous groups of nephrons can be identified only by the marking technique, except for those at Th 7 level, whose distance from the caudal pole does not alter up to the 11th embryonic day.

Animals

Histochemical detection of sugar residues in the chick embryo mesonephros with lectin-horseradish peroxidase conjugates.

Fragments of mesonephros were taken from chick embryos and studied from the 4th to the 21st day of incubation. A battery of seven different horseradish peroxidase-labelled lectins was used to study the distribution of carbohydrate residues in glycoconjugates along the mesonephric nephron during the period of excretory activity and the period of involution. ConA and WGA reacted at every site of the nephron thus showing the ubiquitous presence of alpha-D-mannose and N-acetyl-D-glucosamine. SBA was a good marker of the proximal tubule. Other lectins, such as PNA and LTA, reacted only for a short time at some sites during the considered period of incubation. The presence of sialic acid was detected in the podocytes, capillary wall and mesangial cells. From the 10th-11th day of incubation changes were noted in the proximal tubule as shown by PNA reactivity. This may be significant as regards the exact stage of incubation during which the involution of mesonephros begins.

Acetylglucosamine

Human testicular development and the role of the mesonephros in the origin of a dual Sertoli cell system.

Some aspects of the development of the human testis (and overy) are discussed and the main theories regarding gonadal differentiation summarized. The major part of this review deals with the origin and differentiation of the three groups of somatic cellular content: Sertoli cells, Leydig cells and peritubular cells. The most important role of the mesonephros in gonadal development is described. Under the influence of the mesonephros, a second type of meiosis-inducing Sertoli cell differentiates and becomes the opponent of a meiosis-preventing type of Sertoli cell which derives from the coelomic epithelium. All somatic cells are pooled in the central gonadal blastema which is part of the medulla. They migrate via the rete blastema to the sites of their final differentiation. Included are the precursors of the Leydig cells and the peritubular cells.

Cell Differentiation

The role of cellular locomotion in leukemic infiltration. An organ culture study on penetration of L 5222 rat leukemia cells into the chick embryo mesonephros.

The significance of cellular locomotion for leukemic infiltration was investigated using L 5222 rat leukemia cells. Previous cinemicrographic studies have shown that these cells are able to locomote only after formation of a uropod-like posterior extension. This characteristic locomotive configuration of L 5222 cells is easily recognizable in scanning electron micrographs and appropriate sections. Leukemia cells were inoculated on slices of chick embryo mesonephros incubated for 24h; at this time the fragments are completely encapsulated. Leukemic infiltration is found to begin within the first 2 h and to increase gradually up to the end of the observation period at 72 h. Spread of leukemia cells occurs mainly in the intertubular spaces; the tubular epithelium is only rarely affected. In all stages of infiltration, L5222 cells with the characteristic locomotive configuration are frequently recorded. Besides this strong although indirect indication for the significance of locomotion, further evidence was provided by experiments performed at 25 degrees C and 18 degrees C. In accordance with the previous cinemicrographic finding that at these temperatures L 5222 cells are unable to produce their posterior extension, no leukemic infiltration mesonephros fragments is recognizable at subnormal temperatures.

Animals

Renal carbonic anhydrase in the quail Coturnix coturnix japonica. II. Changes of enzyme activity in developing and regressing mesonephros.

Carbonic anhydrase activity was studied during development and regression of the quail mesonephros by in situ and extra situm investigation. A close correlation was noted between enzyme expression and tissue morphofunctional state. Carbonic anhydrase appears in early development; its highest activity is reached when the kidney is actively secreting, followed by a decrease concomitant with tissue involution. The main localization of the reaction product is the distal tubule showing strongly positive cells intercalated with clear, negative ones. In the functional organ, staining was found at the level of transitional and connecting segments and Wolffian duct. The comparison with the histochemical pattern of the quail metanephros suggests that the functional meaning of renal carbonic anhydrase might be the same both in transitory and in permanent kidney.

Animals

Scanning electron microscopy of the renal corpuscle of the mesonephros in the lamprey, Entosphenus japonicus Martens.

The renal corpuscle of the lamprey mesonephros was studied under the scanning electron microscope. Bowman's capsules with individual spaces are chockshaped sacs closely packed together along a medial artery. The lateral walls of the capsules are apposed to those of neighbouring capsules. Glomerular capillaries from the medial artery extend radially between the apposed walls of neighbouring Bowman's capsules. Bulgings of capillaries into the capsular space are associated with mesangial folds of the capsular epithelium. The transitional zone of the visceral layer with podocytes and the parietal layer of squamous epithelium is bounded by linearly arranged rod-shaped epithelial cells. Apertures of the urinary tubule are lined by cells equipped with a fascicle of cilia.

Animals

Histochemical localization of Mg2+-Na+-K+-adenosine triphosphatase in different stages of the sheep mesonephros.

Mesonephroi of sheep embryos ranging from 12 to 100 mm C.R. length were examined for the occurrence and localization of transport-ATPase. Native cryostat sections were incubated according to the technique of Guth and Albers for demonstrating the nitrophenylphosphatase activity of Mg2+-Na+-K+-adenosine triphosphatase. The basal cytoplasm of the collecting tubule of the narrow segment of the distal tubule exhibit strong activity, the wide segment of the distal tubule is moderately active. Glomeruli, proximal tubule, and Wolffian duct remain unstained. The basal labyrinths of the reactive nephron segments are believed to be the sites of a Na+-K+ exchange pump. In mature and regressing mesonephroi, the findings fully agree with biochemical data; in maturating mesonephroi, whose basal labyrinth is not yet fully established, the biochemical assay proves to be more sensitive. The specifity of the reaction was ascertained by diverse inhibitors and activating ions. The localization of Mg2+-ATPase is different to the above mentioned reaction pattern, as it shows moderate activity in the proximal tubule, too (mature mesonephros). Mesonephroi of very young embryos exhibit strongest Mg2+-ATPase activity in the proximal tubule; here the distal and collecting tubule stain only moderately.

Adenosine Triphosphatases

Scanning electron microscopical observations on the differentiating mesonephros of the chick embryo.

Chick embryos were staged according to the method of Hamburger and Hamilton [1951] and fixed. Cross sections through the cephalic fourth of the mesonephric ridges were examined by scanning electron microscopy. The steps in glomerular differentiation could be observed with ease. The first foot processes to appear in podocytes arose directly from the basal surface of the cell body. In a second step, lateral branches appeared and gave off secondary or even tertiary branches that interdigitated with those from neighbouring podocytes, following a pattern that was very similar to the one previously described by other authors in metanephric nephrons. Endothelial pores appeared in the glomerular capillaries at very early stages of the glomerular differentiation. The differentiation of the epithelium of proximal tubules was characterized by the growth of apical microvilli and of finger-like evaginations from the lateral membranes. At stages 20 and 21, the most differentiated glomeruli had only basal foot processes; only after stage 25 did the first generation nephrons reach full maturity. Because during this period the mesonephros is known to produce urine, our results indicate that nephrons start to function before they have completed their differentiation.

Animals

Pronephros and mesonephros--Cohnheim revisited.

Erroneous quotations in the literature and Cohnheim's statement, in his Lectures on General Pathology, that the Wolffian body or mesonephros is the first anlage of the urogenital system prompted this description of the growth of our knowledge of the early development of the kidney. Some of the pertinent older literature is reviewed, and the concept of the holonephros, as opposed to the trinephric view of kidney development, is discussed. Emphasis is placed on the decreasing functional significance of the pronephros with evolutionary development, to the extent that the role of the pronephros in the human is questioned. Cohnheim's seemingly erroneous reference to the development of the kidney is shown to be merely a reflection of the views current at his time.

History, 19th Century

[Development of the nephrons of the mesonephros of chicken embryo].

The number of nephron populations in the postinduction period was established in 6- and 8-days chicken embryos and the development of an individual nephron and its parts was studied. The investigation by microdissection method has shownand the number of nephrons is different along the length of the kidney. Only two layers ofthe nephrons were found in the cranial portion, while in the caudal direction their number increased up to 4-6 populations which distinguished from one another by the glomerule position, the length of the nephron and its segments. All the populations of the ventral nephrons enter immediately into the mesonephritic (Wolffian) duct, while the dorsal nephrons have a system ofcollecting tubes by which they are connected with the mesonephric duct. The development of mesonephros was accompained by the increase of the absolute length of the nephrons of all populationsand their segments. Laboratory of Individual Development, Institute of Physiology, Czechoslovakian Academy of Sciences, Prague, and Laboratory of the Evolution of the Kidney and Water-Salt Exchange, Sechenov Institue of Evolutionary Physiologyand Biochemistry, Leningrad.

Animals

Development of the mesonephros, metanephros and gonads in the foetal dog.

We describe the chronology of the development of the renal system and gonads of 19 greyhound foetuses. The smallest specimens were embedded in paraffin wax and were sectioned serially at 10 microns; the rest were sectioned at 50 microns in an LKB 2250 PMV cryomicrotome. Selected sections were stained with haematoxylin and eosin. The mesonephros, metanephros and gonads were measured in drawings of the sections, using the area fraction programme of a Vids II image analyse.

Animals

Detection of sex-specific proteins in chick embryo gonads and mesonephros: effects of estradiol benzoate or tamoxifen on their expression.

Gonadal and mesonephric protein patterns from 19 day old normal chick embryos were investigated by two-dimensional polyacrylamide gel electrophoresis. Under these conditions, several sex-specific polypeptides were detected. As concerns gonadal extracts, four sex-specific polypeptides, all restricted to the cytosol, were present in the testis, whereas three sex-specific polypeptides, two localized in the cytosol, the other being membrane-bound, were identified in the ovary. Among the ovary-specific polypeptides two proved to be estrogen-dependent. They appeared in the left testis of embryos after early estradiol benzoate treatment and their expression was reduced in the ovary after early exposure to the antiestrogen, tamoxifen. Mesonephros extracts of both sexes also differed in their protein composition since three additional polypeptides (one in both the cytosolic and membrane fractions, the others in the cytosol) not found in females were found to be present in males. None appeared to be affected after either estradiol or tamoxifen treatment.

Animals