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Biomedical subjects

L Barajas

Publications and source records attributed to L Barajas.

At least 19 recordsLinked to original sources

Evidence for NOS-containing renal neuronal somata transiently expressing a catecholaminergic phenotype during development in the rat.

Transiently catecholaminergic cells (TC-cells) expressing tyrosine hydroxylase (TH) have been shown in a variety of tissues during embryonic life. To investigate the possible relationship of nitric oxide synthase (NOS)-containing renal neuronal somata (RNS) and the TC-cells, we examined serial 100 microm slices of whole kidneys for TH-immunofluorescence and NADPH-d histochemistry during prenatal and postnatal development. The number of TH-cells increased during the prenatal period, peaked at birth and were very rare by PD21. A subpopulation of TH-immunoreactive RNS displayed NADPH-d activity. By PD21 the TH-positive RNS had practically disappeared while the number of NADPH-d positive RNS was markedly increased. These results suggest that kidneys possess transient catecholaminergic cells which display NOS-activity and that NOS expression may be the end-point in the differentiation of the RNS.

Animals↗

Omphalocele with absent radial ray (ORR): a case with diploid-triploid mixoploidy.

We observed omphalocele, absence of radii, hypoplasia of one humerus, a hemivertebra, and syndactyly in a stillborn male at 22 weeks of gestation. Craniofacial and genitourinary abnormalities were absent. DNA measurement by flow cytometry on a paraffin-embedded autopsy specimen showed 32% triploid cells. ORR (omphalocele-radial ray) complex appears to be a consistent combination, and diploid-triploid mixoploidy may be one of its causes.

Abnormalities, Multiple↗

Clinically silent primary adrenal lymphoma: a case report and review of the literature.

Primary adrenal lymphoma (PAL) is extremely uncommon. We describe a case of clinically silent non-Hodgkin's B-cell lymphoma of diffuse large cell type with exclusive left adrenal localization. The tumor was discovered by computed tomography (CT) as a 2.5-cm dense mass and diagnosed at autopsy. Literature concerning this unusual neoplasm is reviewed. During the early stage, particularly when the lesion is small, PAL is likely to be missed. This unusual entity should be included in the differential diagnosis of adrenal masses so that early diagnosis may be made and intervention might dramatically affect the clinical outcome.

Adrenal Gland Neoplasms↗

RT-PCR study of the distribution of connexin 43 mRNA in the glomerulus and renal tubular segments.

An RT-PCR study of the distribution of connexin 43 (Cx43) mRNA in glomeruli and along the rat tubular segments was carried out to establish the differential expression of Cx43 in the different segments of the tubule, in renal regions, in isolated glomerular preparations (IGP), and in microdissected glomeruli. The mRNA level of Cx43 in macrodissected renal regions appeared in the following order: inner papilla > outer papilla and IGP > outer medulla and cortex. Among the microdissected tubules, inner medullary collecting ducts (IMCD) expressed the highest level of Cx43 mRNA, followed by the cortical collecting ducts (CCD). The proximal convoluted tubules and proximal straight tubules expressed significantly less Cx43 than the IMCD, glomeruli, and CCD. Medullary thick ascending limb and distal convoluted tubules showed the lowest level of Cx43 mRNA. The RT-PCR results of the microdissected segments correlate well with those obtained by RT-PCR of the renal regions. The high concentration of Cx43 mRNA in the IMCD together with the observation of abundant punctate immunofluorescence for Cx43 suggests that the IMCD not only expresses Cx43 mRNA but also that the mRNA is translated to Cx43 protein.

Animals↗

Transforming growth factor alpha-immunoreactivity in neural tissues of the rat stomach.

We report TGF alpha immunoreactivity in neurons of the myenteric plexus and in nerve fibers in the muscle and submucosal layers of the rat stomach. Association of TGF alpha staining nerve fibers to vessels and smooth muscle cells gives morphological evidence that EGF/TGF alpha's actions to increase mucosal blood flow and gastric motility may be mediated by TGF alpha derived from neural structures. These data suggest that TGF alpha plays a role in the neural control of the gastric function.

Animals↗

Mice lacking transforming growth factor alpha have an increased susceptibility to dextran sulfate-induced colitis.

BACKGROUND & AIMS: There is indirect evidence that transforming growth factor alpha (TGF-alpha) is an important mediator of mucosal defense and repair. TGF-alpha knockout mice and TGF-alpha-deficient mice (wa-1) provide novel approaches to evaluate the role of TGF-alpha in preserving the integrity of the colon. METHODS: Colitis was induced by oral administration of dextran sodium sulfate (DSS, 5 g/dL) to knockout mice, their genetic controls (GC), wa-1 mice, and BALB/c mice. TGF-alpha was also administered intraperitoneally to wa-1 mice to evaluate the effect of exogenous TGF-alpha in DSS colitis. RESULTS: In response to DSS, nearly 60% of the entire colonic mucosa was destroyed in knockout and wa-1 mice, compared with 22% in GC mice and 16% in BALB/ c mice. Body weight loss was doubled in knockout (28%) and wa-1 mice (23%) compared with GC (11%) and Balb/c mice (12%). TGF-alpha application to wa-1 mice reduced the severity of mucosal injury by almost 70% compared with controls. CONCLUSIONS: The marked susceptibility of TGF-alpha knockout and wa-1 mice to DSS and the obvious amelioration of the colonic injury by exogenous TGF-alpha application in wa-1 mice suggest that TGF-alpha is a mediator of protection and/or healing mechanisms in the colon.

Animals↗

Cell-specific protein and gene expression in the juxtaglomerular apparatus.

1. The juxtaglomerular apparatus (JGA) consists of a tubular component, the macula densa (MD), attached to a vascular component consisting of the afferent and efferent arterioles and the extraglomerular mesangium. The JGA is richly innervated by sympathetic fibres. 2. The MD is morphologically, histochemically and functionally different from the ascending thick portion of the loop of Henle where it is located. 3. The vascular component includes the vascular smooth muscle cells of the arteriole, the renin-producing cells or juxtaglomerular cells, extraglomerular mesangial cells (Goormaghtigh cells) and endothelial cells. They are coupled by gap junctions. 4. Physiological evidence indicates that the composition of tubular fluid at the MD regulates renin secretion and glomerular haemodynamics and that the JGA is important in the maintenance of body salt-water homeostasis. Evidence suggests that the MD exerts its action on the vascular component through a paracrine mechanism.

Animals↗

Distribution of nitric oxide synthase-containing ganglionic neuronal somata and postganglionic fibers in the rat kidney.

Nitric oxide synthase (NOS)-immunoreactive neurons were identified in the rat kidney by using an antibody against type Ia NOS and the avidin-biotin complex immunoperoxidase method in whole kidneys examined in 100 microns serial sections. The histochemical method for demonstration of the nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was also used to characterize NOS-containing neurons. All somata showing NOS immunoreactivity also displayed NADPH-d activity. The greatest number of neuronal somata were observed in groups at the wall of the renal pelvis and in the angular space formed by the pole of the renal parenchyma and renal pelvic wall. They were also seen at the renal hilus close to the renal artery and along the interlobar vasculature. The size of the neuronal somata in the 35-day-old rat ranged from 13.6 to 34.8 microns, with a mean size of 21.52 +/- 4.81 microns. Seventy percent, however, ranged in size from 17.8 to 26.8 microns. The shape of the neuronal somata also varied, with the majority having an ovoid or round shape. The distribution of the postganglionic fibers was investigated by means of the camera lucida. Postganglionic fibers projected into the wall of the renal pelvis and/or to the interlobar arteries extending to the arcuate arteries and to the beginning of the afferent arterioles. The NOS-immunoreactive neurons may have a vasodilator and relaxing function on the renal pelvic wall and vasculature. In addition, the presence of NOS-containing nerve fibers in nerve bundles, which are known to have predominantly vasomotor and sensory fibers, suggest that they may have a possible modulatory role on renal neural function.

Animals↗

Development of NOS-containing neuronal somata in the rat kidney.

An investigation of the changes in size, number and distribution of NOS-containing neuronal somata in the rat kidney was undertaken. The immunoperoxidase method for the staining of NOS and the histochemical method for the demonstration of NADPH-d were applied to serial thick sections (100 microns) of whole kidneys. Animals at embryonic day 14 (ED14), ED16, ED18, ED20, at birth (PD0), and at postnatal days 4 (PD4), PD12, PD21 and PD35 were studied. NOS-containing neuronal somata were observed by the 20th day of gestation in some kidneys and were consistently seen at birth. They were usually seen in groups of separated neuronal somata or in tight clusters. The neuronal somata were often attached or embedded in nerve bundles. As the kidney developed, the number of neuronal somata separated from each other increased, while the number of clusters remained relatively constant. The size of the neuronal somata increased with development. There were highly significant statistical differences in the size of the neuronal somata between all groups, except between PD12 and PD21. The distribution of neuronal somata at birth was similar to that of the adult. They could be found, (a) at the free renal pelvic wall; (b) in the connective tissue at the angular space between the renal pelvis and the renal parenchyma (SPP); and (c) along the interlobar vessels. At birth and in the early stages of development, the greatest number of neuronal somata were located at the renal pelvis. In the later stages of development, more neuronal somata appear in the connective tissue between the renal pelvis and the renal parenchyma. The location of NOS-containing neuronal somata suggests that they might have a modulatory role on the sympathetic and sensory renal nerves all through development.

Animals↗

Bile ductule formation in fetal, neonatal, and infant livers compared with extrahepatic biliary atresia.

The cell of origin of intrahepatic bile ducts during fetal development remains a subject of controversy, although there has been recent evidence that they form from hepatocytes. However, the origin of neoductules and ducts in the setting of liver disease has not been extensively investigated in humans. Using anticytokeratins characteristic of hepatocytes and bile ducts, we repeated earlier studies of fetal development to compare ductule formation in normal developing and newborn livers with the ductules formed during extrahepatic biliary atresia. We utilized an antibody to proliferating cell nuclear antigen (PCNA) staining to determine which cells were in active DNA synthesis (S phase) during fetal development and liver disease progression. The results indicated that hepatocytes undergo a phenotypic switch (metaplasia) to form ductular cells during fetal development. There was no ductular cell replication in the fetal livers. In contrast, both bile ductular metaplasia and proliferation were observed in biliary atresia. Therefore, both a limiting plate phenotypic switch to ductules and replication of ductular cells play a role in the increase in the ductules seen in the progression to biliary cirrhosis. Bile ductular proliferation in biliary atresia, however, was less than that seen in hepatocytes, whereas the number of bile ductules increased and the relative proportion of hepatocytes diminished as the accompanying periductular fibrosis progressed to cirrhosis.

Bile Ducts↗

Colocalization of NADPH-diaphorase and dopamine beta-hydroxylase in the neuronal somata of the rat kidney.

Neuronal somata in the rat kidney are very often part of ganglionated plexus and contain nitric oxide synthase (NOS). Examining serial 100 microns slices of whole kidneys, we identified three subpopulations of neuronal somata by: (a) staining for NADPH-diaphorase (NADPH-d) histochemistry followed by the demonstration of dopamine beta-hydroxylase (DBH) by immunoperoxidase, and (b) staining for DBH by immunofluorescence followed by the demonstration of NADPH-d histochemical activity. The largest subpopulation of neuronal somata displayed both DBH immunoreactivity and NADPH-d histochemical activity. The second largest group of somata showed NADPH-d activity only. A small group of neuronal somata showed only DBH immunoreactivity. The presence of catecholaminergic characteristics in NOS-containing neuronal somata is unusual and raises the question as to their origin. Their heterogeneity suggests different functions for the different subpopulations.

Adrenergic Fibers↗

Rat connexins 30.3 and 31 are expressed in the kidney.

Six connexin genes have previously been shown to be expressed in the rat kidney. Given the structural and functional diversity of the kidney, we hypothesize that other connexin genes may be expressed. We have partially screened a rat kidney cDNA library using low-stringency hybridization conditions with cDNA probes from rCx 43 and rCx 26 and report here the isolation of two connexin cDNA clones, rCx 30.3 and rCx 31, that have not previously been shown to be expressed in the rat kidney. Furthermore, rCx 30.3 utilizes two distinct transcripts in the kidney, while rCx 31 utilizes two transcripts in skin but only one in the kidney.

Amino Acid Sequence↗

Localization of connexin43 in rat kidney.

The localization of connexin43 (Cx 43) in rat kidney was investigated by the indirect immunofluorescence technique with polyclonal antisera raised against Cx 43. Cx 43 is a gap junction protein expressed in a variety of tissues. The typically punctuated gap junction immunofluorescence (GJI) was observed in the renal arterial and arteriolar system. In the renal artery the GJI was concentrated in the media. In the juxtamedullary nephrons, the GJI is particularly abundant in the vascular bundles. There is abundant GJI in the extraglomerular mesangium while in the afferent arteriole GJI appears decreased. Abundant GJI was observed in the inner medullary collecting ducts and pelvic epithelium. The localization of Cx 43 immunofluorescence observed in this study is only in partial agreement with the results of ultrastructural investigations on the distribution of gap junctions in the kidney. An extensive tight junctional system has been demonstrated in the collecting duct system. However, gap junctions have been reported to be absent. Further studies to resolve this discrepancy are required.

Animals↗

Renal tubular dysgenesis: evidence of abnormality in the renin-angiotensin system.

Renal tubular dysgenesis is an autosomal recessive condition characterized by short, abnormally developed cortical tubules that lack proximal differentiation. Despite the lack of normal proximal tubules, the major site of water resorption in the kidney, the principal clinical manifestations are caused by fetal and neonatal oliguria. The kidneys in three cases of neonatal renal tubular dysgenesis were found to contain large amounts of immunohistochemically reactive renin in preglomerular arterioles, glomerular hilums, and glomerular mesangial areas, far exceeding the intensity of staining and the numbers of sites stained in control kidneys. The increased accumulation of renin may reflect strong local vasoconstriction, which is responsible for reduced glomerular perfusion. This accumulation suggests faulty feedback control of renin secretion, the basis of which is still to be identified.

Case-Control Studies↗

Nitric oxide synthase immunoreactive neurons in the rat kidney.

The presence of neurons with nitric oxide synthase (NOS) immunoreactivity was investigated in the rat kidney. Whole kidneys were examined by means of serial sections. The indirect immunocytochemical technique using polyclonal antibody raised against rat brain type Ia NOS and the histochemical technique for nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase are used in this study. NOS-immunoreactive (NOS-IR) neurons varied in size and were observed: (1) associated with nerve bundles at the hilus of the kidney, (2) in the proximity of the lower or middle portion of the interlobar arteries, and (3) on the wall of the renal pelvis. We are presenting anatomic evidence for the presence of neurons in the rat kidney. Their location is consistent with the existence of a parasympathetic innervation of the rat kidney.

Amino Acid Oxidoreductases↗

The rat renal nerves during development.

The prenatal and postnatal development of the innervation of the rat kidney has been investigated using immunocytochemical methods. The efferent innervation was studied using dopamine-beta-hydroxylase and neuropeptide Y antibodies. Calcitonin gene related peptide and substance P antibodies were used to investigate the afferent innervation. Kidneys from embryos of 14 to 20 days, from newborn rats, and from animals of 4, 10, 12, 21, 38, 60, and 90 days of age were studied. Slices of whole kidneys were analyzed, and frozen sections were used to investigate the location of the nerves in more detail. Both afferent and efferent nerves are observed inside the kidney by embryonic day 16. At birth, the afferent nerves are found (1) forming a rich plexus in the renal pelvis; (2) associated with the renal vasculature as far as the interlobular arteries (cortical radial arteries) and (3) in the corticomedullary connective tissue. The efferent innervation appears, at birth, to extend to the interlobular arteries and to the afferent arterioles of the perihilar juxtamedullary nephrons. The efferent innervation increases rapidly during the following days, and by postnatal day 21 a distribution of the innervation similar to that of the adult is observed. While the afferent innervation reaches the major target regions of the kidney by birth, the efferent does most of its expansion into the kidney postnatally. Afferent and efferent fibers are found, extrarenally and intrarenally, in the same nerve bundles. This proximity between afferent and efferent fibers may represent anatomical bases for their interaction in the adult as well as during development.

Afferent Pathways↗

The renal nerves in the newborn rat.

Immunocytochemical methods were used to investigate the distribution of afferent [calcitonin gene-related peptide-(CGRP) immunoreactive and substance P-immunoreactive] nerves and efferent (neuropeptide Y-immunoreactive and dopamine beta-hydroxylase-immunoreactive) nerves in the kidneys of rats within the 1st day of life. The newborn rat kidney possesses an afferent and efferent innervation. Both afferent and efferent nerves reach the kidney in the same bundles. The afferent sensory fibers predominate overwhelmingly in the renal pelvis and ureter while the efferent fibers clearly predominate in the vasculature. The corticomedullary connective tissue contains both types of innervation with a more prominent afferent innervation (CGRP immunoreactive). Only afferent arterioles of perihilar nephrons were innervated by efferent sympathetic fibers. The distribution and extent of afferent and efferent innervation is consistent with the renal nerves playing a significant role in the transition from fetal to newborn life. The close proximity between afferent and efferent fibers suggests a possible interaction between the two systems.

Afferent Pathways↗

Lack of control of renin release by adrenergic nervous system in the aglomerular toadfish.

Aglomerular toadfish, Opsanus tau, release renin in response to hemorrhage or vasodilator drugs, presumably by stimulating a renal arterial baroreceptor. We aimed to determine whether the adrenergic nervous system and prostaglandins play a role in the control of renin release in unanesthetized toadfish kept in 50% seawater. Isoproterenol (1 microgram/kg) increased plasma renin activity (PRA) fourfold and decreased blood pressure (BP); both effects were abolished by a concomitant infusion of propranolol. Propranolol itself slightly decreased the basal level of heart rate and BP, but not that of PRA. Norepinephrine (1 microgram/kg) increased BP, but did not change PRA. Repeated injection of 6-hydroxydopamine did not alter resting levels of either PRA or BP. Monoamine-specific nerve fluorescence activity could not be demonstrated in association with arterioles of kidneys from intact toadfish or from those treated with monoamine oxidase inhibitor and norepinephrine (5 mg/kg). Furthermore, treatment of toadfish with indomethacin (10 or 20 mg/kg) prevented neither the increase in PRA nor the reduction in BP after a massive hemorrhage. These results indicate that renin release in toadfish primarily occurs in response to a reduction in renal arterial pressure, whereas it appears unlikely that the adrenergic nervous system or prostaglandins have a significant role in the control of renin release.

Angiotensin II↗