Immunohistochemical localization of a calcium pump and calbindin-D28k in the oviduct of the laying hen

Wasserman, R. H.; Smith, Christina; Smith, Carissa M.; Brindak, M. E.; Fullmer, Curtis S.; Krook, L; Penniston, John T.; Kumar, Rajiv

doi:10.1007/bf00315999

Cited by 60 publications

(21 citation statements)

References 39 publications

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“…In the shell gland the calcium transporter PMCA was localized in the apical membranes of tubular gland cells, which is in agreement with previous reports [7,35]. Staining intensity and localization of PMCA was unaffected by estrogen treatment in both hybrids.…”

Section: Discussionsupporting

confidence: 92%

Exogenous estradiol improves shell strength in laying hens at the end of the laying period

et al. 2014

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BackgroundCracked shells, due to age related reduction of shell quality, are a costly problem for the industry. Parallel to reduced shell quality the skeleton becomes brittle resulting in bone fractures. Calcium, a main prerequisite for both eggshell and bone, is regulated by estrogen in a complex manner. The effects of estrogen, given in a low continuous dose, were studied regarding factors involved in age related changes in shell quality and bone strength of laying hens. A pellet containing 0.385 mg estradiol 3-benzoate (21-day-release) or placebo was inserted subcutaneously in 20 birds each of Lohmann Selected Leghorn (LSL) and Lohmann Brown (LB) at 70 weeks of age. Eggs were collected before and during the experiment for shell quality measurements. Blood samples for analysis of total calcium were taken three days after the insertion and at sacrifice (72 weeks). Right femur was used for bone strength measurements and tissue samples from duodenum and shell gland were processed for morphology, immunohistochemical localization of estrogen receptors (ERα, ERβ), plasma membrane calcium ATPase (PMCA) and histochemical localization of carbonic anhydrase (CA).ResultsEstrogen treatment increased shell thickness of both hybrids. In addition, shell weight and shell deformation improved in eggs from the brown hybrids. The more pronounced effect on eggs from the brown hybrid may be due to a change in sensitivity to estrogen, especially in surface epithelial cells of the shell gland, shown as an altered ratio between ERα and ERβ. A regulatory effect of estrogen on CA activity, but not PMCA, was seen in both duodenum and shell gland, and a possible connection to shell quality is discussed. Bone strength was unaffected by treatment, but femur was stronger in LSL birds suggesting that the hybrids differ in calcium allocation between shell and bone at the end of the laying period. Plasma calcium concentrations and egg production were unaffected.ConclusionsA low continuous dose of estrogen improves shell strength but not bone strength in laying hens at the end of the laying period.

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Section: Discussionsupporting

confidence: 92%

Exogenous estradiol improves shell strength in laying hens at the end of the laying period

et al. 2014

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“…This Ca 2+ export is extremely rapid during calcification and corresponds to a consumption of the total plasmatic Ca 2+ pool every 12 min. Studies of Ca 2+ transfer in vivo using perfusion of uterus [8,9] and in vitro exploring the effects of inhibitors of ion ATPases or carbonic anhydrase [10,31], and ionic analysis of uterine fluid during eggshell formation [5], made it possible to build a first model of Ca 2+ transfer in the uterus (Figure1): Ca 2+ , HCO 3 - secretion and Na + reabsorption was considered to occur against their electrochemical gradient, to involve active intracellular transfer as shown by specific inhibitors [8-10] and to occur in the uterine glandular cells as revealed by immunohistochemistry of transport proteins [32]. Trans-epithelial transfer of Ca 2+ occurs in three steps as observed in all transporting epithelia: Ca 2+ influx through a downhill gradient, an intracellular Ca 2+ transport involving calbindin 28 kDa protein [33] and active output into the lumen through a Ca 2+ pump [4].…”

Section: Discussionmentioning

confidence: 99%

“…Ca 2+ secretion towards the uterine fluid occurs via an active process, involving the Ca 2+ ATPase [7,32,43]. This has recently been associated with the PMCA4 (plasma membrane ATPase Ca 2+ ) [16].…”

Section: Discussionmentioning

confidence: 99%

Identification of uterine ion transporters for mineralisation precursors of the avian eggshell

et al. 2012

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BackgroundIn Gallus gallus, eggshell formation takes place daily in the hen uterus and requires large amounts of the ionic precursors for calcium carbonate (CaCO3). Both elements (Ca2+, HCO3-) are supplied by the blood via trans-epithelial transport. Our aims were to identify genes coding for ion transporters that are upregulated in the uterine portion of the oviduct during eggshell calcification, compared to other tissues and other physiological states, and incorporate these proteins into a general model for mineral transfer across the tubular gland cells during eggshell formation.ResultsA total of 37 candidate ion transport genes were selected from our database of overexpressed uterine genes associated with eggshell calcification, and by analogy with mammalian transporters. Their uterine expression was compared by qRTPCR in the presence and absence of eggshell formation, and with relative expression levels in magnum (low Ca2+/HCO3- movement) and duodenum (high rates of Ca2+/HCO3- trans-epithelial transfer). We identified overexpression of eleven genes related to calcium movement: the TRPV6 Ca2+ channel (basolateral uptake of Ca2+), 28 kDa calbindin (intracellular Ca2+ buffering), the endoplasmic reticulum type 2 and 3 Ca2+ pumps (ER uptake), and the inositol trisphosphate receptors type 1, 2 and 3 (ER release). Ca2+ movement across the apical membrane likely involves membrane Ca2+ pumps and Ca2+/Na+ exchangers. Our data suggests that Na+ transport involved the SCNN1 channel and the Na+/Ca2+ exchangers SLC8A1, 3 for cell uptake, the Na+/K+ ATPase for cell output. K+ uptake resulted from the Na+/K+ ATPase, and its output from the K+ channels (KCNJ2, 15, 16 and KCNMA1).We propose that the HCO3- is mainly produced from CO2 by the carbonic anhydrase 2 (CA2) and that HCO3- is secreted through the HCO3-/Cl- exchanger SLC26A9. HCO3- synthesis and precipitation with Ca2+ produce two H+. Protons are absorbed via the membrane’s Ca2+ pumps ATP2B1, 2 in the apical membrane and the vacuolar (H+)-atpases at the basolateral level. Our model incorporate Cl- ions which are absorbed by the HCO3-/Cl- exchanger SLC26A9 and by Cl- channels (CLCN2, CFTR) and might be extruded by Cl-/H+ exchanger (CLCN5), but also by Na+ K+ 2 Cl- and K+ Cl- cotransporters.ConclusionsOur Gallus gallus uterine model proposes a large list of ion transfer proteins supplying Ca2+ and HCO3- and maintaining cellular ionic homeostasis. This avian model should contribute towards understanding the mechanisms and regulation for ionic precursors of CaCO3, and provide insight in other species where epithelia transport large amount of calcium or bicarbonate.

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“…There is no detectable CaBP -9k in the gland or uterine lumen indicating that none is released from the gland apex with the calcium ions. In the chicken shell gland, an analogous organ to the uterus, an active Ca ++ ATPase localised by immunocytochemistry on the gland cell apex (Wasserman et al 1991) has been suggested to mediate the calcium transport out of the gland cell which shows a uniFigs. 36-39.…”

Section: Discussionmentioning

confidence: 99%

Calcium transport and the localisation of calbindin-D 9k in the ruminant placenta during the second half of pregnancy

Wooding

Morgan

Jones

et al. 1996

Cell and Tissue Research

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In late pregnancy the sheep fetus requires 3 g of calcium per day, all of which must be transported across the trophoblast epithelium of the placenta. Such high levels of calcium transport across other epithelia are normally associated with the presence of calbindin-D9 or -28k. Our immunocytochemical results show that ovine, bovine and caprine interplacentomal trophoblast have high levels of calbindin-D9k, about eight to ten times more than in the placentomal region. The protein is detectable only in the uninucleate trophoblast cells in sheep and goat, the frequent binucleate cells show none. The calbindin-D9k is also present in the maternal glandular epithelium but not the surface epithelium of the uterus. The cellular distribution of the calbindin-D9k immunoreactivity suggests a soluble protein homogenously distributed through cytosol and nucleoplasm but absent from all organelles and intercellular spaces. In contrast, the uterine milk protein(s) are localised in Golgi cisternae and secretory vesicles in gland cells and in apical small endocytic vesicles and lysosomes in the uninucleate trophectodermal cells. The distribution of calbindin-D9k supports the concept that it mediates the high calcium flux by facilitated diffusion and not via any vesicular, membrane-bounded system.

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Immunohistochemical localization of a calcium pump and calbindin-D28k in the oviduct of the laying hen

Cited by 60 publications

References 39 publications

Exogenous estradiol improves shell strength in laying hens at the end of the laying period

Exogenous estradiol improves shell strength in laying hens at the end of the laying period

Identification of uterine ion transporters for mineralisation precursors of the avian eggshell

Calcium transport and the localisation of calbindin-D 9k in the ruminant placenta during the second half of pregnancy

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