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Journal ArticleDOI

H3 receptor renal expression in normal and diabetic rats

07 Mar 2015-Inflammation Research (Springer Basel)-Vol. 64, Iss: 5, pp 271-273
TL;DR: H3R is expressed in the apical membrane by collecting duct cells in the kidney of rats and it is significantly increased in diabetic animals, suggesting its involvement in fluid homeostasis.
Abstract: To extend our previous observation of H4R upregulation in the kidney of diabetic rats, we evaluated in the same specimens the presence of the H3R. Kidney specimens from 24 8-week-old male Wistar rats (12 non-diabetic and 12 diabetic animals) were processed for both immunohistochemical and immunofluorescence analyses. H3R is expressed in the apical membrane by collecting duct cells in the kidney of rats and it is significantly increased in diabetic animals. These data support the hypothesis that H3R could also mediate non-neuronal histamine effects, suggesting its involvement in fluid homeostasis.

Summary (2 min read)

Introduction

  • Recently, their group demonstrated the presence of the histamine H4 receptor (H4R) in resident renal cells of the loop of Henlé and its profound upregulation in the kidney of diabetic rats[1].
  • In renal transplant patients the urinary and blood levels of histamine are elevated.
  • In the kidney of diabetic rats, histamine was increased compared with controls[2].
  • Basically, histamine has been reported to regulate the renal microcirculation, to increase salt and water excretion[3-5], decrease the ultrafiltration coefficient by reducing the total filtration surface area[4], and increase renin release[6].
  • The aim of this study was to extend their previous observation on H4R in healthy and diabetic rats evaluating in the same specimens the renal expression of H3R.

Materials and Methods

  • Animals, protocols, diabetes induction, biochemical and histological parameters have been previously reported[1].
  • Conventional immunohistochemical procedures were employed as described.
  • Renal sections were incubated overnight with anti-H3R (349–358) (2 µg/ml)[7], followed by a three-layer streptavidin–biotin–peroxidase complex staining method.
  • After counterstaining with DAPI, photomicrographs were obtained by Apotome systems at x40 objective.
  • Values are expressed as mean ± SEM positive cell/collecting duct percentage of individual rats (ten images/zone each) from the different experimental groups.

Results

  • The immunohistochemical analysis (Fig. 1A) revealed a clear H3R-like immunoreactivity in control rats, predominantly in the renal medulla and the papilla.
  • In contrast, in the cortex the immunoreactivity was generally very low, and no signal was detected in the glomeruli.
  • Notably, when diabetic and non-diabetic animals were compared, a significant increase in immunoreactivity, suggestive of H3R receptor upregulation, was observed (Fig 1B).
  • The above data, indicating a higher H3R-like immunoreactivity in the apical membrane of epithelial tubular cells mosty located in medulla and papilla, suggested a collecting duct expression profile for the receptor.
  • To confirm this result, immunoflorescence co-staining was performed with AQP2, expressed in the apical membrane by collecting duct cells in the kidney.

Discussion

  • The authors demonstrate for the first time that the H3R is mainly expressed in the apical membrane by collecting duct cells of the rat, and that this protein is significantly upregulated in the kidneys of diabetic animals.
  • The authors histochemical data add the collecting duct cells to the growing list of non-neuronal H3R-expressing cells already reported[7-9], thus providing further evidence for a role of H3R in mediating non-neuronal histamine effects.
  • The collecting duct plays a pivotal role in kidney function and homeostasis by regulating ions and water transport.
  • The ability to concentrate urine is impaired in conditions such as diabetes insipidus, Histamine, whose levels have been reported to be increased in the kidney of diabetic animals[2], has been shown to increase salt and water excretion[8-10].
  • Whether this receptor has a compensatory or pathological implication and its role as a pharmacological target in diabetic nephropathy remains to be established.

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10 August 2022
AperTO - Archivio Istituzionale Open Access dell'Università di Torino
Original Citation:
H3 receptor renal expression in normal and diabetic rats
Published version:
DOI:10.1007/s00011-015-0808-y
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This version is available http://hdl.handle.net/2318/158681 since 2016-11-11T15:28:57Z

The final publication is available at Springer via
http://www.springerlink.com/openurl.asp?genre=article&id=doi:10.1007/
s00011-015-0808-y

H
3
receptor renal expression in normal and diabetic rats.
Alessandro Pini
1*
, Paul L Chazot
2*
, Eleonora Veglia
3
, Aldo Moggio
3
, Arianna Carolina Rosa
3
*
Authors contributed equally to this work
1
Dipartimento di Medicina Sperimentale e Clinica Sezione di Anatomia e Istologia Università degli Studi di Firenze,
Largo Brambilla 3, 50134 Florence, Italy;
2
School of Biological and Biomedical Sciences and Wolfson Research
Institute, Durham University, South Road, Durham DH1 3LE, UK;
3
Dipartimento di Scienza e Tecnologia del
Farmaco, Università di Torino, Via P. giuria 9, 10125 Turin, Italy;
Corresponding authors Arianna Carolina Rosa, PhD
Dipartimento di Scienza e Tecnologia del Farmaco, University of Turin, Italy, Via P.
Giuria 9, 10125, Turin, Italy
Phone: +390116707955
Fax: +390116707688
e-mail: ariannacarolina.rosa@unito.it

Abstract
Introduction
In order to extend our previous observation of H
4
R upregulation in the kidney of diabetic rats, we evaluated in the same
specimens the presence of the H
3
R.
Materials and methods
Kidney specimens from 24 8-week-old male Wistar rats (12 non-diabetic and 12 diabetic animals) were processed for
both immunohistochemical and immunofluorescence analyses.
Results and conclusion
H
3
R is expressed in the apical membrane by collecting duct cells in the kidney of rats and it is significantly increased in
diabetic animals. These data support the hypothesis that H
3
R could also mediate non-neuronal histamine effects,
suggesting its involvement in fluid homeostasis.
Key Words histamine H
3
receptor, histamine, kidney, diabetes, collecting ducts

Introduction
Recently, our group demonstrated the presence of the histamine H
4
receptor (H
4
R) in resident renal cells of the loop of
Henlé and its profound upregulation in the kidney of diabetic rats[1]. This observation adds to independent evidence of
a role for histamine in renal (patho)physiology. In healthy subjects the administration of loading doses of L-histidine led
to an increase of histamine concentration in urine. In renal transplant patients the urinary and blood levels of histamine
are elevated. In the kidney of diabetic rats, histamine was increased compared with controls[2]. Basically, histamine has
been reported to regulate the renal microcirculation, to increase salt and water excretion[3-5], decrease the ultrafiltration
coefficient by reducing the total filtration surface area[4], and increase renin release[6].
The aim of this study was to extend our previous observation on H
4
R in healthy and diabetic rats evaluating in the same
specimens the renal expression of H
3
R.
Materials and Methods
Animals, protocols, diabetes induction, biochemical and histological parameters have been previously reported[1].
Conventional immunohistochemical procedures were employed as described. Immunoperoxidase staining for H
3
R was
performed on 5 µm kidney sections for formalin-fixed tissue from 24 male 8-week-old Wistar rats (12 non-diabetic and
12 diabetic animals). Renal sections were incubated overnight with anti-H
3
R (349358) (2 µg/ml)[7], followed by a
three-layer streptavidinbiotinperoxidase complex staining method. Photomicrographs were acquired randomly with a
digital camera connected to a light microscope equipped with a x40 objective (Leica DM750). Images were processed
by ImageJ 1.41 (NIH, USA) software and quantified using the Color Deconvolution image analysis tool. The per-
centage area was calculated for H
3
R-positive tissue. Values are mean ± SEM of the optical density (in arbitrary units)
measurements of individual rats (ten images/zone each) from the different experimental groups. For
immunofluorescence and confocal analysis, after antigen retrieval and blocking, kidney sections were incubated with
primary anti-H
3
R and anti-AQP2, antibody, followed by incubation with corresponding Alexa Fluor secondary
antibodies. After counterstaining with DAPI, photomicrographs were obtained by Apotome systems (Zeiss) at x40
objective. The percentage of positive cells for H
3
R and AQP2 was determine by ImageJ 1.41 software. Values are
expressed as mean ± SEM positive cell/collecting duct percentage of individual rats (ten images/zone each) from the
different experimental groups.
To confirm the absence of false staining, tissue was also screened in the absence of primary antibodies and following
pre-incubation with (349-358) peptide (data not shown). All sections were immunostained in a single session to
minimize artifactual differences.

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Abstract: This study compared the cardiac sympatho-inhibitory responses produced by agonists at α2 -adrenergic (B-HT 933), dopamine D2 -like (quinpirole) and histamine H3 /H4 (immepip) receptors between normoglycaemic and streptozotocin-pretreated (diabetic) pithed rats. Intravenous (i.v.) continuous infusions of B-HT 933, quinpirole or immepip were used in normoglycaemic and diabetic pithed rats to analyse their sympatho-inhibitory effects on the electrically-stimulated cardioaccelerator sympathetic outflow. Both in normoglycaemic and diabetic animals, B-HT 933 (until 100 μg/kg per minute) and quinpirole (until 10 μg/kg per minute) inhibited the tachycardic responses to electrical sympathetic stimulation, but not those to i.v. bolus of exogenous noradrenaline. These sympatho-inhibitory responses were more pronounced in diabetic than in normoglycaemic animals. Accordingly, the areas under the curve for 100 μg/kg per minute B-HT 933 and 10 μg/kg per minute quinpirole in diabetic rats (1065 ± 70 and 920 ± 35, respectively) were significantly smaller (P < .05) than those in normoglycaemic rats (1220 ± 45 and 1360 ± 42, respectively). In contrast, immepip infusions produced cardiac sympatho-inhibition in normoglycaemic (until 10 μg/kg per minute), but not in diabetic (until 100 μg/kg per minute) animals. Our results suggest that in diabetic pithed rats: (i) the more pronounced cardiac sympatho-inhibition to B-HT 933 and quinpirole may be probably due to up-regulation of α2 -adrenergic and dopamine D2 -like receptors, respectively; (ii) the histamine H3 /H4 receptors do not seem to play a sympatho-inhibitory role; and (iii) there is a differential participation of α2 -adrenergic and dopamine D2 -like receptors, which may certainly represent therapeutic targets for the treatment of diabetic complications such as cardiovascular autonomic neuropathy.

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4 citations


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References
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TL;DR: The structure and function of the key transporters and the complex interplay of regulatory factors that modulate principal cell ion and water transport are addressed.
Abstract: The principal cell of the kidney collecting duct is one of the most highly regulated epithelial cell types in vertebrates. The effects of hormonal, autocrine, and paracrine factors to regulate principal cell transport processes are central to the maintenance of fluid and electrolyte balance in the face of wide variations in food and water intake. In marked contrast with the epithelial cells lining the proximal tubule, the collecting duct is electrically tight, and ion and osmotic gradients can be very high. The central role of principal cells in salt and water transport is reflected by their defining transporters-the epithelial Na(+) channel (ENaC), the renal outer medullary K(+) channel, and the aquaporin 2 (AQP2) water channel. The coordinated regulation of ENaC by aldosterone, and AQP2 by arginine vasopressin (AVP) in principal cells is essential for the control of plasma Na(+) and K(+) concentrations, extracellular fluid volume, and BP. In addition to these essential hormones, additional neuronal, physical, and chemical factors influence Na(+), K(+), and water homeostasis. Notably, a variety of secreted paracrine and autocrine agents such as bradykinin, ATP, endothelin, nitric oxide, and prostaglandin E2 counterbalance and limit the natriferic effects of aldosterone and the water-retaining effects of AVP. Considerable recent progress has improved our understanding of the transporters, receptors, second messengers, and signaling events that mediate principal cell responses to changing environments in health and disease. This review primarily addresses the structure and function of the key transporters and the complex interplay of regulatory factors that modulate principal cell ion and water transport.

235 citations

Journal ArticleDOI
TL;DR: Downregulation of cAMP-dependent PKA/ERK1/2/Elk-1 phosphorylation (by activation of H3R) is important in the inhibition of cholangiocyte growth in liver diseases.

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TL;DR: A member of the Gi family, most likely Gi3, is involved in the cAMP-triggered targeting of AQP2-bearing vesicles to the apical membrane of kidney epithelial cells.

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TL;DR: Using a hydrodynamic theory describing transport of macromolecules through an isoporous membrane, it was showed that histamine did not affect glomerular pore size but did produce a major reduction in the ratio of pore area:pore length, a measure of por density, which suggests that the fall in K0 is due largely to a functional reduction in total filtration surface area.
Abstract: We made direct measurements of pregiomerular, glomerular, and postglomerular pressures and flows before and during intra-aortic infusion of a mildly vasodepressor dose of histamine in 20 Munich-Wistar rats. As with whole kidney GFR, single nephron (SN) GFR remained unchanged during histamine infusion, despite significant mean increases in total renal and glomerular plasma flow (QA) rates, as well as in mean glomerular capillary hydraulic pressure (Pec)- These hemodynamic changes were accompanied by proportionately greater reductions in afferent than in efferent arteriolar resistances. A declinein the glomerular capillary ultrafiltration coefficient, Kr, served to offset the increases in QA and PGC. thus accounting for the near-constancy of SNGFR and GFR. Infusion of diphenhydramine, but not metiamide, largely prevented these histamine-induced changes in the glomerular microcirculation, indicating that, in the Munich-Wistar rat, the action of histamine on the glomerulus is channeled largely through an Hi-receptor system. Fractional clearances of uncharged dextrans with molecular radii of 20-42 A, measured in a separate group of five rats, were unaffected by histamine infusion. Using a hydrodynamic theory describing transport of macromolecules through an isoporous membrane, we showed that histamine did not affect glomerular pore size but did produce a major reduction in the ratio of pore area:pore length, a measure of pore density. Taken in conjunction with the finding of a histamine-induced decline in Kr, the results with dextrans suggest that the fall in K0 is due largely to a functional reduction in total filtration surface area. Ore Res 45: 737- 745, 1979

48 citations

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TL;DR: Cell surface biotinylation experiments confirmed that AQP4 is internalized after 20 min of histamine exposure, which may account for the downregulation of water transport, the first evidence for short term rearrangement of OAPs in an established AQP 4-expressing cell line.
Abstract: To test the involvement of the water channel aquaporin (AQP)-4 in gastric acid physiology, the human gastric cell line (HGT)-1 was stably transfected with rat AQP4. AQP4 was immunolocalized to the basolateral membrane of transfected HGT-1 cells, like in native parietal cells. Expression of AQP4 in transfected cells increased the osmotic water permeability coefficient (Pf) from 2.02 +/- 0.3 x 10-4 to 16.37 +/- 0.5 x 10-4 cm/s at 20 degrees C. Freeze-fracture EM showed distinct orthogonal arrays of particles (OAPs), the morphological signature of AQP4, on the plasma membrane of AQP4-expressing cells. Quantitative morphometry showed that the density of OAPs was 2.5 +/- 0.3% under basal condition and decreased by 50% to 1.2 +/- 0.3% after 20 min of histamine stimulation, mainly due to a significant decrease of the OAPs number. Concomitantly, Pf decreased by approximately 35% in 20-min histamine-stimulated cells. Both Pf and OAPs density were not modified after 10 min of histamine exposure, time at which the maximal hormonal response is observed. Cell surface biotinylation experiments confirmed that AQP4 is internalized after 20 min of histamine exposure, which may account for the downregulation of water transport. This is the first evidence for short term rearrangement of OAPs in an established AQP4-expressing cell line.

38 citations

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In this paper, the presence of the histamine H4 receptor ( H4R ) in resident renal cells of the loop of Henlé and its profound upregulation in the kidney of diabetic rats was investigated.