Renal architecture of the dogfish Scyliorhinus caniculus (Chondrichthyes,Elasmobranchii)

Summary The excretory portion of the opisthonephric kidney of Scyliorhinus caniculus displays a mesial zone that is supplied with venous blood by the renal portal system and with arterial blood from the efferent arterioles of the glomeruli, and a zone of lateral bundles that is irrigated with arterial blood via arterioles in parallel to the afferent arterioles of the glomeruli. Each single nephron performs two large convolutions in the mesial tissue and two hairpin loops in the bundle. The nephron is differentiated into renal corpuscle (located between the two zones), neck segment (in the bundle), proximal segment I (beginning in the bundle, major convolution between the zones), proximal segment II (exclusively in the mesial zone), intermediate segment (beginning in the mesial tissue and ending in the bundle), distal segment (exclusively in the bundle) and collecting tubule (beginning in the bundle, with a large convolution in the mesial tissue and ending in the bundle) that joins the collecting duct-ureter system. In the bundles proximal and distal nephron segments, the end of the renal tubule and a central bundle vessel are arranged together and form a complex countercurrent system that is enclosed in a sheath of connective tissue. The bundles provide the structural basis for the creation of an environment with low urea concentration around the final portion of the renal tubules, which is consistent with previous experimental evidence of a significantly lower urea content of the bundles as compared with the blood and the mesial tissue in another marine elasmobranch, Raja erinacea. This condition is thought to lead to passive reabsorption of urea from the fluid of the end of the renal tubule. Separation of individual nephrons in the bundle zone appears to be correlated with the peculiar secondary structure that results from the folding of the bundles and may be in addition a requirement in conjunction with intermittent function of the glomeruli. The zonation of the renal tissue with formation of bundles with counter-current systems is characteristically found in marine Elasmobranchs and is considered to be the morphological correlate to the physiological ability of the marine Elasmobranchii to use urea for osmoregulation.     

FGF8 is required for cell survival at distinct stages of nephrogenesis and for regulation of gene expression in nascent nephrons

During kidney morphogenesis, the formation of nephrons begins when mesenchymal nephron progenitor cells aggregate and transform into epithelial vesicles that elongate and assume an S-shape. Cells in different regions of the S-shaped body subsequently differentiate into the morphologically and functionally distinct segments of the mature nephron. Here, we have used an allelic series of mutations to determine the role of the secreted signaling molecule FGF8 in nephrogenesis. In the absence of FGF8 signaling, nephron formation is initiated, but the nascent nephrons do not express Wnt4 or Lim1, and nephrogenesis does not progress to the S-shaped body stage. Furthermore, the nephron progenitor cells that reside in the peripheral zone, the outermost region of the developing kidney, are progressively lost. When FGF8 signaling is severely reduced rather than eliminated, mesenchymal cells differentiate into S-shaped bodies. However, the cells within these structures that normally differentiate into the tubular segments of the mature nephron undergo apoptosis, resulting in the formation of kidneys with severely truncated nephrons consisting of renal corpuscles connected to collecting ducts by an abnormally short tubular segment. Thus, unlike other FGF family members, which regulate growth and branching morphogenesis of the collecting duct system, Fgf8 encodes a factor essential for gene regulation and cell survival at distinct steps in nephrogenesis.     

Microscopic anatomy of the triton kidney

The structure of the nephron, length of all its segments and the renal architectonics as a whole have been studied in the newt (Triturus vulgaris L.) by means of microdissection and microinjection methods. The microinjection of latex or lissamine green is performed into the blood vessels and nephrons. In one kidney 85-95 nephrons are counted, their number is not the same along the kidney length and increases in the caudal direction. There are nephrons of the ventral, intrarenal and medial populations. The length of the former is 2.7 times as great as that of the latter. A relative length of the nephron segments changes slightly. In all the ventral nephrons a nephrostome is detected. A specific peculiarity of the newt nephron that differs it from that in other vertebrates is the presence of a long distal segment, heterogeneous by its structure. Superficial canaliculi of the kidney are strictly oriented: medially to the renal corpuscles there are loops of the first part of the distal segment, they are vascularized out of the efferent veins system; laterally to the renal corpuscles the loops of the proximal, connective and second part of the distal segments are localized; they receive their blood from some branches of the afferent vein.     

The fine structure of the elasmobranch renal tubule: intermediate, distal, and collecting duct segments of the little skate.

Sharks, skates, and rays (Elasmobranchii) have evolved unique osmoregulatory strategies to survive in marine habitats. These adaptations include a complex renal countercurrent system for urea retention. The fine structure of the complete renal tubular epithelium has yet to be elucidated in any species of cartilagenous fish. The present study, which is a companion to our recent paper describing the ultrastructure of the neck and proximal segments of the elasmobranch nephron, uses thin sections and freeze-fracture replicas to elucidate the fine structural organization of the intermediate, distal, and collecting duct segments of the little skate, Raja erinacea, renal tubule. The epithelium of the intermediate, distal, and collecting duct segments consists of two major cell types: nonflagellar cells, the major epithelial cell type; and flagellar cells, described elsewhere. The intermediate segment consists of six subdivisions lined by cuboidal-columnar cells with variously elaborated microvilli and interdigitations of lateral and basal cell plasma membranes, as well as some subdivisions with distinctive vesicles and granules. The distal segment consists of two subdivisions, both of which are lined by a simple epithelium, and are distinguished from each other by their distinctive contents; dense bodies and granules. The collecting duct segment also has two subdividions, the first lined by a simple columnar epithelium and the second by a stratified columnar epithelium. Both subdivisions have apical secretory granules. The present findings show a more highly specialized and diverse epithelium lining the renal tubule of these cartilagenous fish than is found in either of the "adjacent" phylogenetic taxa, Agnatha or Ostheichthyes, suggesting significant differences among these groups in transepithelial transport mechanisms and renal function.     

Structure of the kidney in the coelacanth Latimeria chalumnae with reference to osmoregulation

The morphology of the nephrons of the coelacanth Latimeria chalumnae was investigated by light microscopy. Each nephron is composed of a large renal corpuscle with well‐vascularized glomerulus, non‐ciliated neck segment, proximal convoluted tubule divided into distinct first and second segments, non‐ciliated intermediate segment, distal tubule, collecting tubule and collecting duct. The parietal layer of the Bowman's capsule of the renal corpuscle is composed of low cuboidal cells. The short non‐ciliated neck segment is lined by cuboidal epithelium. The first and second proximal segments display a prominent brush border and contain amorphous material in their lumen. The second proximal segment differs from the first segment in having taller columnar epithelium and a relatively narrow lumen. The intermediate segment is lined by non‐ciliated columnar epithelium and its lumen appears empty. The distal tubule is narrow in diameter and its cuboidal epithelium is devoid of intercalated cells. A unique feature of L. chalumnae is having binucleate cells in the tubule and collecting duct epithelium. The renal arteries have poorly developed tunica media and its cells contain granular material. The structure of L. chalumnae nephrons correlates well with their osmoregulatory function and resembles those of euryhaline teleosts.     

中华鳖肾脏的组织化学特性研究

利用光镜组织化学反应对中华鳖肾单位的结构和组织化学特性进行了详细的观察和分析。结果表明,中华鳖肾脏为分叶形的实质器官,肾小叶由被膜和实质组成,实质无髓质和皮质之分,但可以区分为外侧区和内侧区。外侧区嗜酸性,主要分布有近端小管和集合管。内侧区呈弱嗜酸性,肾小体、颈段、中间段和远端小管主要分布在内侧区。肾小球PAS反应呈阳性,但其琥珀酸脱氢酶(SDH)弱阳性,碱性磷酸酶(ALPase)、Na+/K+-ATPase和阿利新兰(AB)反应为阴性。足细胞酸性磷酸酶(ACPase)反应呈阳性。近端小管刷状缘嗜伊红,PAS反应以及ALPase、ACPase和Na+/K+-ATPase酶反应呈阳性,而SDH弱阳性。中间段、远端小管、集合管弱嗜酸性,SDH阳性。中间段Na+/K+-ATPase弱阳性。远端小管细胞侧面呈PAS阳性,腔面显示AB阳性。集合管胞质含有许多ACPase阳性颗粒,腔面呈PAS强阳性,AB阳性。甲苯胺兰(TB)染色可见集合管腔面有阳性颗粒,肾小管上皮含有亮、暗两种细胞。上述组化反应和分布结果表明,鳖的肾小管细胞类型较多,近端小管在原尿的重吸收中起主要作用,远端小管和集合管具有分泌黏液作用。中华鳖肾单位的结构与组化特性不仅与哺乳类和鸟类有一定差异,也与其他爬行动物不完全相同。       

Chemical and morphological differences in the kidney zones of the elasmobranch, Raja erinacea mitch

The histological investigation of the kidney of the skate Raja erinacea revealed a thin cap of dorsal bundles, which contain segments of single nephrons that are arranged separately in a countercurrent manner, and a large ventral zone, where the second proximal segments (PII) and parts of the lower nephron are located. This zonation is apparent in fresh, unfixed material and makes it possible to separate small tissue samples under a dissecting microscope. The osmolality in both zones does not differ. The dorsal bundle zone had a lower urea concentration and a higher sodium concentration than the ventral zone. The differences in the mean concentrations of the tissue samples indicate a gradient for the two substances along the bundles. Determinations of amounts of water and solutes per mg solute-free, dry tissue of the two zones, showed that the amounts of water, total osmolytes, Na and K were greater in the bundle zone than in the ventral zone, while the amount of urea was identical in the two zones. This indicates that the lower urea concentration in the bundle zone is established through an accumulation of Na and water in the interstitium. The countercurrent arrangement of very early and late segments of single renal tubules supports the concept of passive reabsorption of urea in the kidney of the marine elasmobranch.     

Kidney regeneration through nephron neogenesis in medaka

Although renal regeneration is limited to repair of the proximal tubule in mammals, some bony fish are capable of renal regeneration through nephron neogenesis in the event of renal injury. We previously reported that nephron development in the medaka mesonephros is characterized by four histologically distinct stages, generally referred to as condensed mesenchyme, nephrogenic body, relatively small nephron, and the mature nephron. Developing nephrons are positive for wt1 expression during the first three of these stages. In the present study, we examined the regenerative response to renal injury, artificially induced by the administration of sublethal amounts of gentamicin in adult medaka. Similar to previous reports in other animals, the renal tubular epithelium and the glomerulus of the medaka kidney exhibited severe damage after exposure to this agent. However, kidneys showed substantial recovery after gentamicin administration, and a significant number of developing nephrons appeared 14 days after gentamicin administration (P < 0.01). Similarly, the expression of wt1 in developing nephrons also indicated the early stages of nephrogenesis. These findings show that medaka has the ability to regenerate kidney through nephron neogenesis during adulthood and that wt1 is a suitable marker for detecting nephrogenesis.     

Use of dual section mRNA in situ hybridisation/immunohistochemistry to clarify gene expression patterns during the early stages of nephron development in the embryo and in the mature nephron of the adult mouse kidney

The kidney is the most complex organ within the urogenital system. The adult mouse kidney contains in excess of 8,000 mature nephrons, each of which can be subdivided into a renal corpuscle and 14 distinct tubular segments. The histological complexity of this organ can make the clarification of the site of gene expression by in situ hybridisation difficult. We have defined a panel of seven antibodies capable of identifying the six stages of early nephron development, the tubular nephron segments and the components of the renal corpuscle within the embryonic and adult mouse kidney. We have analysed in detail the protein expression of Wt1, Calb1 Aqp1, Aqp2 and Umod using these antibodies. We have then coupled immunohistochemistry with RNA in situ hybridisation in order to precisely identify the expression pattern of different genes, including Wnt4, Umod and Spp1. This technique will be invaluable for examining at high resolution, the structure of both the developing and mature nephron where standard in situ hybridisation and histological techniques are insufficient. The use of this technique will enhance the expression analyses of genes which may be involved in nephron formation and the function of the mature nephron in the mouse.     

Morphology of the kidney of adult bowfin, Amia calva, with emphasis on "renal chloride cells" in the tubule

The nephron of adult bowfin, Amia calva, was described using light and electron microscopic techniques. The kidney of the bowfin possesses an abundant supply of renal corpuscles with each consisting of a glomerulus and a Bowman's capsule of visceral (podocyte) and parietal layers. No juxtaglomerular apparatus is present. The epithelium of the tubule is continuous with the parietal epithelium and is divisible in descending order into neck, first proximal, second proximal, first distal, second distal, and collecting segments. The tubules drain into a complex system of collecting ducts that ultimately unite with the main excretory duct, the archinephric duct. Mucous cells are the dominant cell throughout the entire ductular system. Nephrostomes are dispersed along the kidney capsule. The neck segment has a ciliated epithelium, and while both proximal segments possess a prominent brush border, the fine structure of the first implies involvement in protein absorption and the second in the transport and reabsorption of solutes. The cells of the first distal segment are characterized by deep infolding of the plasma membrane and a rich supply of mitochondria suggesting the presence of a mechanism for ion transport. The second distal segment is composed of cells resembling the chloride cells of fishes and these cells are present in progressively decreasing numbers in the collecting segment and duct system so that only a few are present in the epithelium of the archinephric duct. The "renal chloride cells" possess an abundant network of smooth tubules and numerous mitochondria with a rich supply of cristae. Glycogen is also a conspicuous component of these cells. The presence of "renal chloride cells" in this freshwater holostean, in other relatively primitive freshwater teleosts, and in larval and adult lampreys is discussed with reference to both phylogeny and the need for a special mechanism for renal ion conservation through absorption.     

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.     

Modification of genital kidney nephrons for sperm transport in a plethodontid salamander,Eurycea longicauda

Salamanders possess kidneys with two distinct regions: a caudal pelvic portion and cranial genital portion. Nephrons of the pelvic region are responsible for urine formation and transport. Nephrons of the genital region transport sperm from testes to Wolffian ducts; however, nephrons of the genital region possess all the same functional regions found in pelvic kidney nephrons that are involved with urine formation and transport (renal corpuscles, proximal tubules, distal tubules, and collecting ducts). Morphological similarities between pelvic and genital regions stimulated past researchers to hypothesize that nephrons of genital kidneys possess dual function; that is, sperm transport and urine formation/transport. Considering size of glomeruli is directly related to the total amount of blood plasma filtered into the Bowman's space, we tested the hypothesis that nephrons of genital kidneys have reduced urine formation function by comparing glomerular size between nephrons of pelvic and genital kidney regions in Eurycea longicauda with general histological techniques. Light microscopy analysis revealed that glomeruli of pelvic kidneys were significantly larger than those measured from genital kidneys. Transmission electron microscopy analysis also revealed modifications in genital kidney nephrons when compared to pelvic kidney nephrons that suggested a decrease in urine formation function in genital kidneys. Such modifications included a decrease in basal and lateral plasma membrane folding in genital kidney proximal and distal tubules compared to that of pelvic kidney proximal and distal tubules. Genital kidney proximal tubules were also ciliated, which was not observed in pelvic kidney proximal tubules. In conclusion, although structurally similar at the histological level, it appears that nephrons of genital kidneys have decreased urine formation function based on glomerular size comparison and nephron ultrastructure.     

Structure Organization of Urinary System in the Yellow Spotted Mountain Newts （Salamandridae： Neurergus microspilotus）

This study deals with the histomorphology of the mesonephros in male and female Neurergus microspilotus. The slender and narrow kidneys are positioned in the retro peritoneal position up against the ventral aspect of vertebral column and may extend the length from the esophagus-stomach junction to cloaca. The kidney in both sexes is composed of sexual（anterior） and pelvic（posterior） parts. The duct of sexual kidney is a narrow duct which is lying alongside its lateral edge. In the female, it is connected to the ureters and then the duct of defi nitive kidney. Before entering the cloaca, two ureters are joined together and open to the apex of the cloaca. In the male, after entering the sexual kidney, the sperm leave the testis through efferent ducts, then these ducts join together and eventually form Bidder＇s duct. The Bidder＇s duct joins the Bowman＇s capsule of the nephrons in the sexual kidney and the nephrons make collecting ducts which are fi lled with both sperm and urine. After leaving the kidney, all the collecting ducts are connected to the Wolffi an duct. Wolffi an duct joins the ureters（merge from defi nitive kidney） just before entering the cloaca. Based on serial paraffi n sections, nephrons consist of a fi ltration unit, the Malpighian corpuscle, and a renal tubule, which can be divided into 4 morphologically distinct segments： proximal tubule（first and second segment）, distal tubule, and collecting tubule. Collecting tubules merge and form a branch system that opens into collecting ducts.     

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.     

Comparative histological, histochemical and ultrastructural studies of the nephron of selected snakes from the Egyptian area

The present study was aimed to compare and contrast the histochemical, histological and ultrastructural variations (microanatomical differences) in the nephrons of selected snake species, Eryx jaculus (Boidae), Psammophis sibilans (Colubridae), Naja haje (Elapidae) and Echis pyramidum (Viperidae) from Egypt. The structural studies were carried out by conventional light and electron microscopy. The nephron, the renal unit of snakes, consists of renal corpuscle, proximal tubule, intermediate segment, distal tubule and collecting tubule. The renal corpuscles have large capillaries with clear and dark fenestrated endothelial cells. The proximal tubule showed long microvilli, cytoplasmic vacuoles, developed endoplasmic reticulum and abundant mitochondria. The intermediate segment was lined by ciliated cells. The lining cells of the distal tubules showed few microvilli, abundant dense mitochondria and clear vesicles of mucous appeared in the terminal portion. The collecting tubules consisted of mucous cells. In summary, the ultra-structure studies of nephrons revealed several interspecies similarities and also some intra-species differences in species of snakes.     

Cysts in cultured fetal rat kidneys.

The formation of cysts has been studied in kidneys removed from day-15 and day-18 rat fetuses and cultured in a mixture of medium 199 (Morgan et al., '50) for periods of up to 15 days. Gas phases of 95% O2 and 5% CO2, and 95% air and 5% CO2, were employed, the latter being considered more satisfactory. In day-15 kidneys cysts formed from the ampullary portions of the collecting tubules after 2 days whereas cysts derived from nephrons were not seen until 9 days of culturing. The latter arose from developing juxtamedullary nephrons. In day-18 kidneys cysts from collecting tubules and nephrons were both present after 3 days of culturing. The latter, in this instance, originated mostly in immature midcortical nephrons, the juxtamedullary mephrons having undergone rapid degeneration. The tubular portion of the nephron seemed to be the primary site of dilatation. Under culture conditions cysts of nephrons thus formed from immature actively developing nephrons and not from those that were mature. Cysts associated with collecting tubules arose from the ampullary (terminal) portions of the latter in both day-15 and day-18 cultured kidneys. The study of cultured mammalian fetal kidneys can provide information on the nature and genesis of renal cysts. It is possible that the same technique also may be helpful for examining the effects of teratogens directly on the kidney.     

Nephron structure and immunohistochemical localization of ion pumps and aquaporins in the kidney of frogs inhabiting different environments

Amphibians inhabit areas ranging from completely aqueous to terrestrial environments and move between water and land. The kidneys of all anurans are similar at the gross morphological level: the structure of their nephrons is related to habitat. According to the observation by light and electron microscopy, the cells that make up the nephron differ among species. Immunohistochemical studies using antibodies to various ATPases showed a significant species difference depending on habitat. The immunoreactivity for Na+,K(+)-ATPase was low in the proximal tubules but high in the basolateral membranes of early distal tubules to collecting ducts in all species. In the proximal tubule, apical membranes of the cells were slightly immunoreactive to H(+)-ATPase antibody in aquatic species. In the connecting tubule and the collecting duct, the apical membrane of intercalated cells was immunoreactive in all species. In aquatic species, H+,K(+)-ATPase immunoreactivity was observed in cell along the proximal, distal tubule to the collecting duct. However, H+,K(+)-ATPase was present along the intercalated cells of the distal segments from early distal to collecting tubules in terrestrial and semi-aquatic species. In the renal corpuscle, the neck segment and the intermediate segment, immunoreactivities to ion pumps were not observed in any of the species examined. Taking together our observations, we conclude that in the aquatic species, a large volume of plasma must be filtered in a large glomerulus and the ultrafiltrate components are reabsorbed along a large and long proximal segment of the nephron. Control of tubular transport may be poorly developed when a small short distal segment of the nephron is observed. On the contrary, terrestrial species have a long and well-developed distal segment and regulation mechanisms of tubular transport may have evolved in these segments. Thus, the development of the late distal segments of the nephron is one of the important factors for the terrestrial adaptation.     

Microanatomy of the renal cortex in the domestic fowl

Two aspects of the avian renal cortical microanatomy previously were unclear. The precise in situ folding patterns and orientations of the nephrons with respect to the other cortical elements had not been demonstrated. It also was not known whether certain nephron segments are supplied exclusively by either the arterial or the portal blood flow. In the present study, a new casting compound was developed to allow selective examination of the cortical components by light microscopy. Cortical nephrons at the surface of the kidney were serially sectioned and reconstructed in order to determine: (a) their relationships to the vasculature and collecting ducts; (b) the location and characteristics of the tubule segments; and (c) the primary and secondary folding patterns of the tubules. The anatomical findings were documented individually and then summarized in a comprehensive diagram of the superficial cortical microanatomy. In addition, an in vivo method was used to determine the extent of portal blood distribution to the nephron segments. It was demonstrated that renal portal blood suffuses all of the segments except for the loops of Henle.     

Phenolsulphotransferase: localization in kidney during human embryonic and fetal development

Summary The aim of our study was to localize phenolsulphotransferase (PST) in the developing mesonephric and metanephric kidneys of the human embryo and fetus using immunohistochemical methods with an antibody preparation recognizing members of the human phenolsulphotransferase enzyme family. In embryonic and early fetal development of the metanephric kidney, PST is located primarily in derivatives of the ureteric bud such as the ureter, pelvis, calyces and collecting ducts. This predominance declines by mid-fetal life: first, as nephrons evolve and develop they become increasingly PST-immunoreactive such that in mature metanephric kidney, the proximal tubules are highly PST-reactive, with other elements of the nephron also immunopositive (albeit at lower reactivities) and secondly, with the formation of an immunonegative transitional epithelium in ureter, pelvis and calyces, the reactivity retained in collecting ducts is only a small proportion of the total. The distribution of PST immunoreactivity is relatively uniform in proximal tubular cells throughout development, in contrast to collecting ducts, where, in fetal life, this reactivity is displaced to apices and bases by intracellular glycogen deposits. Mesonephric kidney tubules and the mesonephric duct are PST-immunoreactive and although mesonephric immunopositivity overlaps with that in the developing metanephric kidney the renal contribution to sulphation is absent or low at a time when the developing conceptus is most vulnerable to the potential toxic effects of teratogens.