Catch me if you can-emission patterns of human bodies in relation to postmortem changes.

The present study examines for the first time the emission patterns and olfactory signatures of 9 complete human corpses of different stages of decomposition. Air sampling was performed inside the body bags with solid sorbents and analysed by coupled gas chromatography-mass spectrometry after thermal desorption (TD-GC-MS). Furthermore, odour-related substances were detected by gas chromatography-olfactometry (GC-O). Sulfurous compounds (mainly dimethyl di- and trisulfide) were identified as most important to the odour perception. Around 350 individual organic substances were detected by TD-GC-MS, notably sulfurous and nitrogenous substances as well as branched alkanes, aldehydes, ketones, alcohols, carboxylic acids, carboxylic acid esters and ethers. A range of terpenes was detected for the first time in a characteristic emission pattern over all decomposition stages. Concentrations of the substances varied greatly, and no correlation between the emission patterns, the stage of decomposition and the cause of death could be found. While previous studies often analysed pig cadavers or only parts of human tissue, the present study shows the importance of analysing complete human corpses over a range of decomposition stages. Moreover, it is shown that using body bags as a kind of "emission test chamber" is a very promising approach, also because it is a realistic application considering the usual transport and store of a body before autopsy.

Altered regional 3D shear wave velocity patterns in youth competitive alpine skiers suffering from patellar tendon complaints - a prospective case-control study.

Patellar tendon (PT) complaints are frequent throughout the population, with increased occurrence in athletes and, particularly, in youth competitive alpine skiers. Timely detection and treatment might improve prospects of recovery. Diagnostic modalities in clinical use to date rely on pain symptoms, manual palpation, and potentially, magnetic resonance imaging (MRI); however, MRI-based imaging yields limited sensitivity. Quantitatively measuring the morphological and mechanical properties of PTs by means of B-mode ultrasound and shear wave elastography (SWE), instead, may allow improved diagnosis or even early detection. We performed B-mode scans and three-dimensional ultrasound shear wave velocity (SWV) mapping and MRI of the PT in 106 youth skiers. A prospective one-year survey on health problems combined with clinical assessments served to categorize symptomatic and asymptomatic youth skiers. Skiers suffering from distal or proximal tendon complaints showed lower SWV in the respective tendon region than asymptomatic skiers (p = 0.035 and p = 0.019, respectively). Youth skiers with distal tendon complaints additionally exhibited decreased SWV in the proximal region compared to asymptomatic counterparts (p = 0.020). Cross-validated analysis of retrospective prediction indicated sensitivity and specificity in detecting tendon complaints in the range of 0.606-0.621 and 0.536-0.650, respectively. MRI detected distal tendon complaints with a sensitivity of 0.410 (12/29) but failed to detect any proximal cases. This study agrees with the most recent literature in that SWE holds promise as a valuable adjunct modality for the diagnosis of PT complaints or even the detection of subclinical prestages. However, to evaluate its prospective predictive value, long-term studies are warranted.Highlights Patellar tendon complaints are a frequent complaint in athletes, particularly in youth competitive alpine skiers, but timely quantitative detection of related tendon properties remains challenging.Quantitative B-mode US and three-dimensional ultrasound shear wave elastography assessments and magnetic resonance imaging were performed in youth competitive alpine skiers.Three-dimensional shear wave elastography was able to discern symptomatic from asymptomatic patellar tendons both in the distal and proximal tendon regions, whereas magnetic resonance imaging failed to detect any proximal cases.

Patellar Tendon Shear Wave Velocity Is Higher and has Different Regional Patterns in Elite Competitive Alpine Skiers than in Healthy Controls.

Competitive alpine skiers are exposed to enormous forces acting on their bodies-particularly on the knee joint and hence the patellar tendon - during both the off-season preparation and in-season competition phases. However, factors influencing patellar tendon adaptation and regional pattern differences between alpine skiers and healthy controls are not yet fully understood, but are essential for deriving effective screening approaches and preventative countermeasures. Thirty elite competitive alpine skiers, all members of the Swiss Alpine Ski Team, and 38 healthy age-matched controls were recruited. A set of two-dimensional shear wave elastography measurements of the PT was acquired and projected into three-dimensional space yielding a volumetric representation of the shear wave velocity profile of the patellar tendon. Multivariate linear models served to quantify differences between the two cohorts and effects of other confounding variables with respect to regional shear wave velocity. A significant (p < 0.001) intergroup difference was found between skiers (mean ± SD = 10.4 ± 1.32 m/s) and controls (mean ± SD = 8.9 ± 1.59 m/s). A significant sex difference was found within skiers (p = 0.024), but no such difference was found in the control group (p = 0.842). Regional SWV pattern alterations between skiers and controls were found for the distal region when compared to the mid-portion (p = 0.023). Competitive alpine skiers exhibit higher SWV in all PT regions than healthy controls, potentially caused by long-term adaptations to heavy tendon loading. The presence of sex-specific differences in PT SWV in skiers but not in controls indicates that sex effects have load-dependent dimensions. Alterations in regional SWV patterns between skiers and controls suggest that patellar tendon adaptation is region specific. In addition to the implementation of 3D SWE, deeper insights into long-term tendon adaptation and normative values for the purpose of preventative screening are provided.

Three-Dimensional Mapping of Shear Wave Velocity in Human Tendon: A Proof of Concept Study.

Ultrasound-based shear wave elastography (SWE) provides the means to quantify tissue mechanical properties in vivo and has proven valuable in detecting degenerative processes in tendons. Its current mode of use is for two-dimensional rendering measurements, which are highly position-dependent. We therefore propose an approach to create a volumetric reconstruction of the mechano-acoustic properties of a structure of interest based on optically tracking the ultrasound probe during free-hand measurement sweeps. In the current work, we aimed (1) to assess the technical feasibility of the three-dimensional mapping of unidirectional shear wave velocity (SWV), (2) to evaluate the possible artefacts associated with hand-held image acquisition, (3) to investigate the reproducibility of the proposed technique, and (4) to study the potential of this method in detecting local adaptations in a longitudinal study setting. Operative and technical feasibility as well as potential artefacts associated with hand-held image acquisition were studied on a synthetic phantom containing discrete targets of known mechanical properties. Measurement reproducibility was assessed based on inter-day and inter-reader scans of the patellar, Achilles, and supraspinatus tendon of ten healthy volunteers and was compared to traditional two-dimensional image acquisition. The potential of this method in detecting local adaptations was studied by testing the effect of short-term voluntary isometric loading history on SWV along the tendon long axis. The suggested approach was technically feasible and reproducible, with a moderate to very good reliability and a standard error of measurement in the range of 0.300-0.591 m/s for the three assessed tendons at the two test-retest modalities. We found a consistent variation in SWV along the longitudinal axis of each tendon, and isometric loading resulted in regional increases in SWV in the patellar and Achilles tendons. The proposed method outperforms traditional two-dimensional measurement with regards to reproducibility and may prove valuable in the objective assessment of pathological tendon changes.

Sensory Perception of Non-Deuterated and Deuterated Organic Compounds.

The chemical background of olfactory perception has been subject of intensive research, but no available model can fully explain the sense of smell. There are also inconsistent results on the role of the isotopology of molecules. In experiments with human subjects it was found that the isotope effect is weak with acetone and D6 -acetone. In contrast, clear differences were observed in the perception of octanoic acid and D15 -octanoic acid. Furthermore, a trained sniffer dog was initially able to distinguish between these isotopologues of octanoic acid. In chromatographic measurements, the respective deuterated molecule showed weaker interaction with a non-polar liquid phase. Quantum chemical calculations give evidence that deuterated octanoic acid binds more strongly to a model receptor than non-deuterated. In contrast, the binding of the non-deuterated molecule is stronger with acetone. The isotope effect is calculated in the framework of statistical mechanics. It results from a complicated interplay between various thermostatistical contributions to the non-covalent free binding energies and it turns out to be very molecule-specific. The vibrational terms including non-classical zero-point energies play about the same role as rotational/translational contributions and are larger than bond length effects for the differential isotope perception of odor for which general rules cannot be derived.

[Magnesium in dogs and cats - physiology, analysis, and magnesium disorders]. / Magnesium bei Hund und Katze ­ physiologische Aspekte, Messung und Störungen im Magnesiumhaushalt.

Magnesium is the second most abundant intracellular cation after potassium. It plays a vital role in almost every metabolic process in the body and is important for bone mineralization, muscle contraction and relaxation, and neuronal signal transduction. Because of its expanding role in intensivecare medicine, there has been a significant increase in knowledge during recent years regarding the functions of magnesium in the body, problems leading to magnesium disorders, and limitations of laboratory testing. Alterations of serum magnesium constitute one of the most prevalent electrolyte abnormalities in critically ill patients and can lead to lifethreatening complications. In addition to human literature, most of the information regarding the role of magnesium is derived from buiatrics. In recent years, a few studies in veterinary medicine have also started to consider the importance of magnesium in dogs and cats.

[Hypermagnesemia of ionized magnesium in 199 dogs - A hospital population-based study on prevalence, etiology and prognosis with special emphasis on kidney disease and the measurement of total magnesium]. / Hypermagnesiämie des ionisierten Magnesiums bei 199 Hunden.

OBJECTIVE: To investigate the prevalence and etiology of an increased concentration of ionized magnesium (iMg) in dogs and to evaluate its prognostic relevance. MATERIALS AND METHODS: From April 2009 to December 2013, serum electrolytes were measured in 9950 dogs using an ion-selective electrode. Inclusion criterion was an iMg concentration ≥ 0.68 mmol/l, whereby total Mg (tMg), potassium, ionized calcium, and micturition were also evaluated in this retrospective study. In the case of repeated measurements in an animal, only the initial measurement of the increased iMg concentration was considered. According to the etiology, patients were categorized in the diagnostic groups: azotemia, iatrogenic/medication associated, endocrine diseases, tissue damage, and unknown etiology of hypermagnesemia. The survival rate was analyzed and the iMg concentration was compared between the diagnostic groups. The correlation between iMg and tMg was assessed. RESULTS: The prevalence of increased iMg concentration was 2.0 % (199/9950). The most prevalent causes were azotemia (80/199; 40 %), followed by iatrogenic hyper-magnesemia (37/199; 19 %), tissue damage (21/199; 11 %), and endocrine diseases (12/199; 6 %). In 49/199 cases (25 %), the etiology of hypermagnesemia was unknown. An additional hypercalcemia was evident in 24/199 dogs (12 %) and 58/199 (29 %) dogs displayed hypocalcemia. In 64 % of the dogs (51/80) with azotemia, underlying renal disease was present, from which 53 % (27/51) exhibited hyperkalemia. Moreover, 37 % (19/51) of the animals with renal disease displayed anuria/oliguria, 59 % (11/19) of which were hyperkalemic. Overall, 83/199 (42 %) of the dogs died, whereby 48/83 (58 %) belonged to the azotemia group. The iMg concentration did not differ significantly between the groups (p = 0.15). There was a poor correlation between iMg and tMg (rs = 0.28). CONCLUSION: An increased iMg concentration is rare and indicative of a severe disease in the majority of cases. Particularly in patients with acute renal disease, hypermagnesemia can be associated with increased mortality. The tMg concentration does not reflect the iMg concentration.

Gender differences in psychopathy links to drug use.

Although the relationship between psychopathic personality traits and substance use has received some attention (Hart & Hare, 1989; Smith & Newman, 1990), gender differences have not been thoroughly assessed. The current study examined whether gender modified the relationship between 2 criminally relevant constructs, (a) psychopathy and its factors and (b) drug use. A sample of 318 participants with criminal histories and recent substance use was assessed for psychopathy using the Psychopathy Checklist: Screening Version and for illicit drug use using the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders. As expected, the impulsive-antisocial traits (Factor 2) of psychopathy were positively related to a number of drug use characteristics (symptoms, age of drug initiation, extent of drug experimentation), whereas the interpersonal-affective traits (Factor 1) showed a negative relationship with drug abuse symptoms and a positive relationship with age of first use. In terms of gender differences, analyses revealed that women showed a stronger association between Factor 1 traits and later age of initiation compared to men, and that Factor 2, and the antisocial facet in particular, were more strongly related to drug abuse in women than men. These findings suggest that psychopathic traits serve as both protective (Factor 1) and risk (Factor 2) correlates of illicit drug use, and Factor 1 may be especially protective in terms of initiation of drug use among women. These conclusions add to the growing literature on potential routes to substance use and incarceration in women.

Seroprevalence of spotted fever group Rickettsiae in dogs in Germany.

Tick-transmitted spotted fever group Rickettsiae (SFGR) are considered emerging disease pathogens in Europe. To assess the situation in Germany, a seroepidemiologic survey of dogs (n=605, which have never left Germany) was conducted to determine the prevalence of antibodies to SFGR. For this purpose, a commercially available enzyme-linked immunosorbent assay (ELISA) test for all SFGR was used. A total of 78% of the dogs were positive for antibodies of the SFGR, and there was a positive correlation between age, tick-infestation, and seropositivity. There was no correlation between clinical state, location, and seropositivity, suggesting that infection with rickettsia is neither related to the health condition of the dogs nor to their abode in Germany. According to our results, all of Germany can be considered as an endemic area for SFGR. This should be taken into account when assessing the epidemiology of human rickettsioses and their impact on public health.

Mouse model of type II Bartter's syndrome. II. Altered expression of renal sodium- and water-transporting proteins.

Bartter's syndrome represents a group of hereditary salt- and water-losing renal tubulopathies caused by loss-of-function mutations in proteins mediating or regulating salt transport in the thick ascending limb (TAL) of Henle's loop. Mutations in the ROMK channel cause type II antenatal Bartter's syndrome that presents with maternal polyhydramnios and postnatal life-threatening volume depletion. We have developed a colony of Romk null mice showing a Bartter-like phenotype and with increased survival to adulthood, suggesting the activation of compensatory mechanisms. To test the hypothesis that upregulation of Na(+)-transporting proteins in segments distal to the TAL contributes to compensation, we studied expression of salt-transporting proteins in ROMK-deficient (Romk(-/-)) mice. Plasma aldosterone was 40% higher and urinary PGE(2) excretion was 1.5-fold higher in Romk(-/-) compared with wild-type littermates. Semiquantitative immunoblotting of kidney homogenates revealed decreased abundances of proximal tubule Na(+)/H(+) exchanger (NHE3) and Na(+)-P(i) cotransporter (NaPi-IIa) and TAL-specific Na(+)-K(+)-2Cl(-)-cotransporter (NKCC2/BSC1) in Romk(-/-) mice, while the distal convoluted tubule (DCT)-specific Na(+)-Cl(-) cotransporter (NCC/TSC) was markedly increased. The abundance of the alpha-,beta-, and gamma-subunits of the epithelial Na(+) channel (ENaC) was slightly increased, although only differences for gamma-ENaC reached statistical significance. Morphometry revealed a fourfold increase in the fractional volume of DCT but not of connecting tubule (CNT) and collecting duct (CCD). Consistently, CNT and CD of Romk(-/-) mice revealed no apparent increase in the luminal abundance of the ENaC compared with those of wild-type mice. These data suggest that the loss of ROMK-dependent Na(+) absorption in the TAL is compensated predominately by upregulation of Na(+) transport in downstream DCT cells. These adaptive changes in Romk(-/-) mice may help to limit renal Na(+) loss, and thereby, contribute to survival of these mice.

Differential localization of vacuolar H+-ATPases containing a1, a2, a3, or a4 (ATP6V0A1-4) subunit isoforms along the nephron.

Vacuolar H(+)-ATPase are multi-subunit containing pumps important for several processes along the nephron such as receptor mediated endocytosis, acidification of intracellular organelles, bicarbonate reabsorption and secretion, and H(+)- extrusion. Mutations in the human a4 (ATP6V0A4) subunit cause distal renal tubular acidosis (dRTA). There are 4 known isoforms of the 'a' subunit (a1-a4). Here we investigated the expression and localization of all four isoforms in mouse kidney. Real-time PCR detected mRNAs encoding all four 'a' isoforms in mouse kidney with a relative abundance in the following order: a4>a2=a1>a3. Immunolocalization demonstrated expression of all 'a' subunits in the proximal tubule and in the intercalated cells of the collecting system. In intercalated cells a1 and a4 isoforms appeared on both the apical and basolateral side and were expressed in all subtypes of intercalated cells. In contrast, a2, and a3 were only found in the apical membrane. a1 and a4 were colocalized in the same cells with AE1 or pendrin, whereas a2 was only found in AE1 positive cells but absent from pendrin expressing intercalated cells. These results suggest that vacuolar H(+)-ATPases containing different 'a' isoforms may serve specific and distinct functions and may help explaining why loss of the a4 isoform causes only dRTA without an apparent defect in the proximal tubule.

Regulation of renal amino acid transporters during metabolic acidosis.

The kidney plays a major role in acid-base homeostasis by adapting the excretion of acid equivalents to dietary intake and metabolism. Urinary acid excretion is mediated by the secretion of protons and titratable acids, particularly ammonia. NH(3) is synthesized in proximal tubule cells from glutamine taken up via specific amino acid transporters. We tested whether kidney amino acid transporters are regulated in mice in which metabolic acidosis was induced with NH(4)Cl. Blood gas and urine analysis confirmed metabolic acidosis. Real-time RT-PCR was performed to quantify the mRNAs of 16 amino acid transporters. The mRNA of phosphoenolpyruvate carboxykinase (PEPCK) was quantified as positive control for the regulation and that of GAPDH, as internal standard. In acidosis, the mRNA of kidney system N amino acid transporter SNAT3 (SLC38A3/SN1) showed a strong induction similar to that of PEPCK, whereas all other tested mRNAs encoding glutamine or glutamate transporters were unchanged or reduced in abundance. At the protein level, Western blotting and immunohistochemistry demonstrated an increased abundance of SNAT3 and reduced expression of the basolateral cationic amino acid/neutral amino acid exchanger subunit y(+)-LAT1 (SLC7A7). SNAT3 was localized to the basolateral membrane of the late proximal tubule S3 segment in control animals, whereas its expression was extended to the earlier S2 segment of the proximal tubule during acidosis. Our results suggest that the selective regulation of SNAT3 and y(+)LAT1 expression may serve a major role in the renal adaptation to acid secretion and thus for systemic acid-base balance.

Expression of heteromeric amino acid transporters along the murine intestine.

Members of the new heterodimeric amino acid transporter family are composed of two subunits, a catalytic multitransmembrane spanning protein (light chain) and a type II glycoprotein (heavy chain). These transporters function as exchangers and thereby extend the transmembrane amino acid transport selectivity to specific amino acids. The heavy chain rBAT associates with the light chain b degrees (,+)AT to form a cystine and cationic amino acid transporter. The other heavy chain, 4F2hc, can interact with seven different light chains to form various transporters corresponding to systems L, y(+)L, asc or x(-)(c). The importance of some of these transporters in intestinal and renal (re)absorption of amino acids is highlighted by the fact that mutations in either the rBAT or b degrees (,+)AT subunit result in cystinuria whereas a defect in the y(+)-LAT1 light chain causes lysinuric protein intolerance. Here we investigated the localization of these transporters in intestine since both diseases are also characterized by altered intestinal amino acid absorption. Real time PCR showed organ-specific expression patterns for all transporter subunit mRNAs along the intestine and Western blotting confirmed these findings on the protein level. Immunohistochemistry demonstrated basolateral coexpression of 4F2hc, LAT2 and y(+)-LAT1 in stomach and small intestine, whereas rBAT and b degrees (,+)AT were found colocalizing on the apical side of small intestine epithelium. In stomach, 4F2hc and LAT2 were localized in H(+)/K(+)-ATPase-expressing parietal cells. The abundant expression of several members of the heterodimeric transporter family along the murine small intestine suggests their involvement in amino acids absorption. Furthermore, strong expression of rBAT, b degrees (,+)AT and y(+)-LAT1 in the small intestine explains the reduced intestinal absorption of some amino acid in patients with cystinuria or lysinuric protein intolerance.

Nongenomic stimulation of vacuolar H+-ATPases in intercalated renal tubule cells by aldosterone.

Renal collecting ducts play a critical role in acid-base homeostasis by establishing steep transepithelial pH gradients necessary for the almost complete reabsorption of bicarbonate and the effective secretion of ammonium into the urine. The mechanisms of urine acidification in collecting ducts involve active, electrogenic hydrogen (H+) secretion and, less importantly, potassium (K+)-H+ exchange. Deranged renal acidification and the inability to lower urine pH are hallmarks of distal tubular acidosis and often result from inborn errors of metabolism involving vacuolar H+-ATPase subunits in the collecting ducts. Three factors regulate H+-ATPase activity in intercalated cells of collecting ducts: the acid-base status, angiotensin II, and aldosterone. Most effects of aldosterone involve activation of the mineralocorticoid receptor and genomic changes in transcription and protein synthesis. Here we demonstrate a nongenomic pathway of vacuolar H+-ATPase activation in intercalated cells of isolated mouse outer medullary collecting ducts (OMCD). In vitro exposure of isolated outer medullary collecting ducts to aldosterone (10 nM) for times as short as 15 min increases vacuolar H+-ATPase activity approximately 2- to 3-fold. Neither inhibition of mineralocorticoid receptors nor of transcription and protein synthesis prevented aldosterone-induced stimulation of H+-ATPase. Incubation with colchicine, however, abolished the stimulatory effect of aldosterone, suggesting a role of the microtubular network for H+-ATPase stimulation. Immunohistochemistry in kidneys from aldosterone-injected mice showed increased apical H+-ATPase staining in OMCD-intercalated cells. The stimulatory effect of aldosterone was associated with a transient rise in intracellular Ca2+ and required intact PKC. Thus, rapid nongenomic modulation of vacuolar H+-ATPase activity in OMCD-intercalated cells by aldosterone may play an additional role in hormonal control of systemic acid-base homeostasis.

Localization and regulation of the ATP6V0A4 (a4) vacuolar H+-ATPase subunit defective in an inherited form of distal renal tubular acidosis.

Vacuolar-type H(+)-ATPases (V-H(+)-ATPases) are the major H(+)-secreting protein in the distal portion of the nephron and are involved in net H(+) secretion (bicarbonate generation) or H(+) reabsorption (net bicarbonate secretion). In addition, V-H(+)-ATPases are involved in HCO(3)(-) reabsorption in the proximal tubule and distal tubule. V-H(+)-ATPases consist of at least 13 subunits, the functions of which have not all been elucidated. Mutations in the accessory ATP6V0A4 (a4 isoform) subunit have recently been shown to cause an inherited form of distal renal tubular acidosis in humans. Here, the localization of this subunit in human and mouse kidney was studied and the regulation of expression and localization of this subunit in mouse kidney in response to acid-base and electrolyte intake was investigated. Reverse transcription-PCR on dissected mouse nephron segments amplified a4-specific transcripts in proximal tubule, loop of Henle, distal convoluted tubule, and cortical and medullary collecting duct. a4 protein was localized by immunohistochemistry to the apical compartment of the proximal tubule (S1/S2 segment), the loop of Henle, the intercalated cells of the distal convoluted tubule, the connecting segment, and all intercalated cells of the entire collecting duct in human and mouse kidney. All types of intercalated cells expressed a4. NH(4)Cl or NaHCO(3) loading for 24 h, 48 h, or 7 d as well as K(+) depletion for 7 and 14 d had no influence on a4 protein expression levels in either cortex or medulla as determined by Western blotting. Immunohistochemistry, however, demonstrated a subcellular redistribution of a4 in response to the different stimuli. NH(4)Cl and K(+) depletion led to a pronounced apical staining in the connecting segment, cortical collecting duct, and outer medullary collecting duct, whereas NaHCO(3) loading caused a stronger bipolar staining in the cortical collecting duct. Taken together, these results demonstrate a4 expression in the proximal tubule, loop of Henle, distal tubule, and collecting duct and suggest that under conditions in which increased V-H(+)-ATPase activity is required, a4 is regulated by trafficking but not protein expression. This may allow for the rapid adaptation of V-H(+)-ATPase activity to altered acid-base intake to achieve systemic pH homeostasis. The significance of a4 expression in the proximal tubule in the context of distal renal tubular acidosis will require further clarification.

Involvement of the MAPK-kinase pathway in the PTH-mediated regulation of the proximal tubule type IIa Na+/Pi cotransporter in mouse kidney.

Reabsorption of phosphate in the proximal tubule is mainly mediated by the type IIa Na(+)/P(i) cotransporter (NaPi-IIa) and tightly regulated by a variety of factors including dietary phosphate intake and parathyroid hormone (PTH). PTH signals through both apical and basolateral PTH receptors and induces the rapid internalization and subsequent degradation of NaPi-IIa. At least two signalling cascades can be activated by PTH: the PLC/PKC and the cAMP/PKA pathways. Recent evidence from OK cell culture suggested the involvement of MAPK kinases in the PTH action. Here we used freshly isolated coronal mouse kidney slices and incubated them in a physiological buffer in the absence and presence of PTH with inhibitors and activators of the various signalling cascades to further study the events leading to internalization of NaPi-IIa. No alterations in the pattern of immunostaining for alpha-tubulin, actin and several brush border membrane proteins demonstrated intactness of the slices over the experimental period. Application of PTH (100 nM) induced a strong decrease of NaPi-IIa brush border staining and internalization after 45 min of incubation. The localization of the Na(+)/sulphate cotransporter (NaSi), however, was not affected. The internalization of NaPi-IIa could be completely prevented by the PKC inhibitor chelerythrine (1 micro M) or the MAPK-kinase (ERK1/2) inhibitor PD098059 (20 micro M). Without PTH both inhibitors alone had no effect. PTH induced phosphorylation of the ERK1/2 MAPK-kinases which was prevented by PD 098059. Separate activation of the cAMP/PKA pathway by 8-Br-cAMP was completely prevented by PD098059 whereas activation of the PLC/PKC pathway by the PKC activator 1,2-dioctanoyl-sn-glycerol (DOG) and the PKG pathway by 8-Br-cGMP induced internalization of NaPi-IIa which could be only partly blocked by PD 098059. Inhibition by SB203580 or activation by anisomycin of the p38 kinase pathway had no influence on NaPi-IIa localization under control conditions or after PTH stimulation. Furthermore, the PTH-induced decrease in NaPi-IIa protein could be reduced by PD 098059. These results suggest that the ERK1/2 MAPK kinase pathway plays a central role in the signalling of PTH leading to specific internalization and subsequent degradation of the type II NaPi-IIa cotransporter in the proximal tubule.