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Laboratory of Comparative Endocrinology, Zoological Institute, Naamsestraat 61, B-3000 Leuven, Belgium
¹ Laboratory of Hormone Biochemistry, Institute of Biochemistry, Academy of Sciences, Uzbekistan

(Requests for offprints should be addressed to C H J Verhoelst; Email: Carla.Verhoelst{at}bio.kuleuven.ac.be)

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
It is widely accepted that type II iodothyronine deiodinase (D2) is mostly present in the brain, where it maintains the homeostasis of thyroid hormone (TH) levels. Although intensive studies have been performed on activity and mRNA levels of the deiodinases, very little is known about their expression at the protein level due to the lack of specific antisera. The current study reports the production of a specific D2 polyclonal antiserum and its use in the comparison of D2 protein distribution with that of type I (D1) and type III (D3) deiodinase protein in the choroid plexus at the blood–brain barrier level. Immunocytochemistry showed very high D2 protein expression in the choroid plexus, especially in the epithelial cells, whereas the D1 and D3 proteins were absent. Furthermore, dexamethasone treatment led to an up-regulation of the D2 protein in the choroid plexus. The expression of D2 protein in the choroid plexus led to a novel insight into the working mechanism of the uptake and transport of thyroid hormones along the blood–brain barrier in birds. It is hypothesized that D2 allows the prohormone thyroxine (T₄) to be converted into the active 3,5,3'-triiodothyronine (T₃). Within the choroidal epithelial cells. T₃ is subsequently bound to its carrier protein, transthyretin (TTR), to allow transport through the cerebrospinal fluid. Neurons can thus not only be provided with a sufficient T₃ level via the aid of the astrocytes, as was hypothesized previously based on in situ hybridization data, but also by means of T₄ deiodination by D2, directly at the blood–brain barrier level.

	Introduction

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It is well established that thyroid hormones play a crucial role in vertebrate development in general and in brain development and maturation in particular. The impact of thyroid hormones (THS) on the development of the central nervous system is shown in situations of clinical or induced TH deficit or excess. Under these circumstances, processes of cell migration and formation of cortical layers are affected in particular, as well as neuronal and glial cell differentiation (Bernal et al. 2003). Thyroxine (T₄), the prohormone produced by the thyroid gland, is considered to be inactive and needs to be converted into the active 3,5,3'-triiodothyronine (T₃). This peripheral deiodination reaction demands the interference of the so-called iodothyronine deiodinases. Three different deiodinases (type I iodothyronine deiodinase (D1), type II iodothyronine deiodinase (D2) and type III iodothyronine deiodinase (D3)) have been cloned and characterized for different species, including mammals, birds, amphibians and fish (Bianco et al. 2002). The three deiodinases differ in the reaction they catalyse, their substrate preferences and their sensitivity to certain inhibitors. D1 catalyzes deiodination of both the outer and the inner ring deiodination (ORD and IRD ) of iodothyronines. D2 and D3 deiodinase are responsible for the maintenance of TH homeostasis in the brain by their respective ORD and IRD activity (Leonard & Visser 1986). Of all the deiodinases, D2 is the one most highly expressed in vertebrate brain. To date, very little is known about the cellular distribution of D2 due to the lack of specific antisera. Diano et al.(2003) described for the first time immunocytochemical evidence that the D2 protein is localized in brain cells, more specifically in astrocytes and tanycytes in the rat hypothalamus. Their results were in agreement with previously reported data on the mRNA distribution of rat D2. Guadaño-Ferraz et al.(1997) reported in situ hybridization studies for D2 in neonatal rat brain. They found excessive D2 mRNA expression in different brain areas, but more importantly, mainly, if not exclusively, in glial cells. It was hypothesized that these support cells would provide a sufficient level of active TH for utilization by the neurons. That way a close coupling would exist between the glial cells and the neurons. Tu et al.(1997) also reported the expression of D2 mRNA in rat hypothalamic tanycytes. Guadaño-Ferraz et al.(1999) confirmed their previous statement by demonstrating that the most TH dependent regions would be protected in a hypothyroid status by an increase of D2 expression in these areas.

In the current study, a specific polyclonal antiserum against chicken D2 was produced and used to compare the D2 protein distribution with that of D1 and D3 proteins in the choroid plexus, a structure which is of extreme importance for TH uptake in the brain via the blood–brain barrier. The main function of the choroid plexus, apart from its protective aspect, is the production of the cerebrospinal fluid by a process that requires the movement of Na⁺, Cl^– and HCO₃^– ions from the blood towards the ventricles. By means of this movement, an osmotic gradient is created and H₂O can be secreted. The epithelial cells of the choroid plexus are equipped with a number of transporter proteins and ion channels in order to fulfil their responsibility in maintaining a homeostasis in the chemical environment of the brain via cerebrospinal fluid production. The monocarboxylate transporter 8 (MCT8) just recently identified by Friesema et al.(2003) to be a very specific TH transporter, is highly expressed in the choroid plexus of mammalian brain (Simpson 2004). The latter indicates that the transport of thyroid hormones in and eventually out of the brain, via the choroid plexus, is indeed of significant importance.

The staining of D2 protein in the choroid plexus was further examined in this study by comparing dexamethasone (DEX) treated and control chicken embryos. DEX is a synthetic glucocorticoid that, upon administration to chicken embryos, increases both brain D2 activity and mRNA levels (Van der Geyten et al. 2001). This experiment allowed further validation of the antiserum. Taken together, the results obtained from the present study gave rise to a novel hypothesis concerning deiodination and transport of thyroid hormones along the blood–brain barrier, namely at the level of the choroid plexus.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Polyclonal antisera production

Based on the full-length amino acid D2 sequence of the chicken, described by Gereben et al.(1999), and taking into account the hydropathy profile of this enzyme and possible cross-reactivity with other deiodinases, a specific peptide was chosen for immunization purposes. The following peptide was synthesized and coupled to keyhole limpet hemocyanin: NH₂-(164)PSDGWAAPGISPPS (179)-COOH. New Zealand White rabbits were injected every 3 weeks and blood collection occurred within 7–10 days after immunization (final bleeding after 8 injections). Serum was separated, snap frozen using a mixture of dry ice and ethanol and stored at –80 °C. The same approach was used for the production of the antisera against chicken D1 and D3, as was described in detail by Verhoelst et al.(2002, 2003). Possible cross-reactivity with the other deiodinases was checked by means of an immunospotting test, in which the three different peptides used for the generation of the respective antisera, namely peptide D1, D3a and the D2 peptide, were spotted on a nitrocellulose membrane. Binding of these peptides to the D2 antiserum was tested using the preimmune D2 antiserum as negative control.

Experiments and tissue preparation

Chicken embryos (Ross, Avibel, Aarschot, Belgium) were injected on day 18 of incubation (E18) with either saline or 50 µg dexamethasone. Four hours after injection the animals were sacrificed and brain tissue was fixed in Bouinhollande solution for 24 h (Sigma). Tissues were washed thereafter for 24 h using running tap water. Before paraffn embedding, the following incubation steps were performed: 50% EtOH (2 h), 70% EtOH (2 h), 95% EtOH (2 h), 100% EtOH (overnight), 1:1 EtOH:xylol (4 h), 100% xylol (4 h), 100% xylol (overnight), 1:1 xylol: paraffn (4 h), 100% paraffn (4 h), 100% paraffn (overnight). Paraffn slices of 7 µm thickness were cut using a historange CBK microtome. The slices were always dried for a minimum 72 h prior to use in immunocytochemical applications.

Immunocytochemical staining

The paraffn slices were hydrated using 100% xylol (2x15 min), 100% EtOH (3 min), 96% EtOH (3 min), 80% EtOH (3 min), 70% EtOH (3 min) and washed in distilled water for 10 secs. Blocking buffer (TBS-Triton X-100+ 5% low fat milk) was applied for 1 h to block the non-specific binding sites. Thereafter, the tissue slices were incubated with a 1:150 dilution of the primary D2 antiserum in TBS-Triton X-100 for 1 h at room temperature and overnight at 4 °C. For the D2 antiserum, both preimmune and exhausted primary antiserum (exhaustion overnight, 4 °C, 200 µg D2 peptide) were tested as negative controls. For the D1 (1:200) and D3 (1:200) antisera, preimmune serum served as a negative control, since exhaustion experiments were already described before (Verhoelst et al. 2002, 2003). The slices were washed twice for 10 min with TBS-Triton X-100. Next, they were incubated with goat-anti-rabbit alkaline phosphatase linked IgG (GAR-AP) (1:250) for 1 h at room temperature. Again, they were washed twice with TBS for 10 min and twice with AP detection buffer (2x10 min). Thereafter, the substrate (NBT/BCIP) was applied (45–60 min). The slices were washed in methanol and enclosed with chromoglycerine.

In situ hybridization

Chicken embryos of 18 days old were perfused with 4% PFA. Subsequently, brains were removed and submitted to the following preparative manipulations: 4% PFA (24 h), PBS (24 h), PBS-10% sucrose (24 h). Finally the brains were put in PBS-10% sucrose solution containing 10% gelatine for embedding and preserved at –80 °C. Cryostate sections of 30 µm were made. Thereafter, the sections were prepared in 4% PFA (10 min), PBS (3x5 min), 0.2N HCl (10 min), TEA buffer (10 min), PBS-1% Triton X-100 (30 min) and PBS (3x5 min). Subsequently, the slices were prehybridized for 2 h at room temperature in hybridization buffer (50% formamide, 10% dextran sulphate, 5% Denhardt’s, 0.625 M NaCl, 0.2 M Na-PIPES pH 6.8 in 50 ml DEPC-AD) containing 0.05 M DTT, 250 µg/µl denatured herring sperm and 250 µg/µl yeast tRNA. Afterwards, the sections were hybridized under coverslips overnight at 80 °C in the same hybridization buffer containing a 1:1000 dilution of a digoxigenin-labeled cRNA probe. The probes were generated by performing PCR using forward primer: 5' (1027)-GCT GGT GGA AGA GTT CTC TGG AGT-10503' and reverse primer: 5' (1262)-GCA CAC TCG CTC AAA TGA AAC CCC-(1285)3'. The thus obtained 260 bp PCR fragment was subsequently subcloned in a PCR II-TOPO plasmid vector (Invitrogen). The sense probe was digested with SacI, while the antisense probe was cut using ApaI, allowing amplification of the cRNA probes using the T7 and SP6 transcription kit (Roche) respectively and DIG-labelled dNTP (Roche). After hybridization, the sections were washed for 45 min at 72 °C in 0.2xsodium saline citrate (SSC), 2x5 min 0.2xSSC at room temperature and 3x5 min in PBS-0.1% Triton X-100 at room temperature. Thereafter, the sections were incubated for 1 h with PBS-BSA-Triton X-100 containing 0.1 M lysine to reduce background signal. Per section, 0.5 ml anti-DIG antibodies (1:5000, in PBS-BSA-Triton X-100) were added and incubation continued overnight at 4 °C. The alkaline phosphatase signal was detected using the NBT/BCIP chromogen system (Roche) according to the manufacturer’s guidelines.

All products used were of the highest quality commercially available. The K U Leuven Ethical Committee for Animal Experiments approved all experimental procedures used.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
An alkaline phosphatase staining with anti-D2 carried out on brain slices of 18-day-old control chicken embryos revealed a very intense staining of the cuboidal epithelial cells in the choroid plexus. The central vascular core of fenestrated capillaries also displayed a very light positive signal (Fig. 1A). However, incubation of the subsequent slice with preimmune serum reveals that this colouring is of a non-specific nature (Fig. 1B). Exhaustion of the primary antiserum using the D2-specific peptide led to the loss of this strong signal in the choroid plexus (Fig. 1C,D).

View larger version (114K): [in this window] [in a new window]   Figure 1 Alkaline phosphatase staining of chicken choroid plexus (E18 SAL) with (A) anti-D2 antiserum, (B) preimmune serum, (C) anti-D2 antiserum, (D) exhausted anti-D2 antiserum, (E) schematic overview of the choroid plexus epithelium. (F-H) Alkaline phosphatase staining of chicken choroid plexus with (F) anti-D2, (G) anti-D1, (H) anti-D3. (I-K) Alkaline phosphatase staining of chicken lateral and third ventricle with (I) preimmune serum, (J-K) anti-D2 antiserum. Arrowheads indicate ependymal cells. (L-M) Comparison of alkaline phosphatase staining of chicken choroid plexus using anti-D2 in (L) control chicken embryos and (M) DEX treated chicken embryos. EpC, epithelial cell; CSF, cerebrospinal fluid; LV, lateral ventricle; IIIrdV, third ventricle; EpdC, ependymal cell.

Since, ontogenetically, the epithelium cells of the choroid plexus originate from the ependymal cells lining the ventricle walls, and since at E18 the chicken brain is still developing, the D2 protein staining was monitored there as well. It can be seen in Fig. 1J–K that the cells circumscribing the lining of the lateral ventricle and those around the third ventricle stained positively using the D2 antiserum in comparison with the preimmune serum (Fig. 1I). These cells are called ependymal cells and tend to be arranged in monolayers of cuboidal to columnar epithelium upon differentiation.

In the dexamethasone injection experiment, a dilution of 1:200 was used for the primary D2 antiserum. Comparing the results of the control animals versus the DEX treated animals shows that DEX treatment causes the D2 protein staining to increase. The cuboidal cells are immuno positive in both cases but the intensity of the AP staining is clearly higher in the DEX treated section (Fig. 1L,M).

In situ hybridization shows the distribution of D2 mRNA in elongated cell clusters throughout the brain. Furthermore, it confirms the presence of D2 mRNA in the epithelial cells of the choroid plexus. Comparison of the negative control slice hybridized with the sense probe (Fig. 2A) with the slice hybridized with the antisense probe (Fig. 2B) reveals a clear D2 mRNA expression.

View larger version (75K): [in this window] [in a new window]   Figure 2 In situ hybridization of the chicken choroid plexus with the (A) sense probe and (B) antisense probe. Closed arrowhead indicates choroid plexus epithelial cells; open arrowhead indicates elongated cell clusters.

View larger version (20K): [in this window] [in a new window]   Figure 3 Scheme illustrating the proposed hypothesis of TH transport along the blood-brain-barrier in birds, compared with the existing scheme in mammals, described by Southwell et al.(1993) (Copyright 1993. The Endocrine Society).

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

However, the data resulting from the current study, namely the expression of the D2 protein in the choroid plexus, implicates the introduction of a new hypothesis concerning TH transport and communication pathways of different neuronal and non-neuronal cells in chicken brain. To better understand this hypothesis, it is of extreme importance to be familiar with the structure and function of the choroid plexus in general. The choroid plexus has a lobulated appearance and consists of a single epithelial cell layer that surrounds the central vascular core. The choroidal epithelial cells originate from the ependymal lining of the ventricles, while the blood vessels and connective tissue that represent the core originate from a vascular fold of the pia mater, namely the tela choroidea. The central core of the choroid plexus contains an unusually large number of capillaries, with very thin endothelial walls that do not possess a blood–brain barrier due to the fact that these capillaries are fenestrated. The epithelial cells of the choroid plexus on the other hand are designed to form a barrier between the blood and the brain. The ependymal cells are polarized, as are many other secretory epithelia, and consist of an apical (ventricle facing) and basolateral (blood facing) membrane (Speake et al. 2001). In mammals, it is described that T₄ enters the epithelial cells of the choroid plexus from the blood via the basolateral membrane. These epithelial cells produce the binding protein TTR, which is known to be a carrier protein for thyroid hormones in general. In mammals, TTR has a higher affinity for T₄ in comparison to T₃ (Schreiber et al. 2002). This is why T₄ binds to the TTR either in the choroidal epithelium or in the cerebrospinal fluid, which allows the transport of the lipophilic TH through the CSF. Thyroxine can reach the subarachnoidal space and can flow towards the rest of the brain coupled to TTR (Fig. 3). It is described that T₄ will be deiodinated by D2 mainly in the astroglial cells (Guadaño-Ferraz et al. 1997). The production of the active T₃ is thus mainly guaranteed by these neuron-supporting cells. Triiodothyronine is then supplied to the neurons, which mainly express D3, for further deiodination and inactivation of the thyroid hormones (Guadaño-Ferraz et al. 1997). In chicken, it has been shown that D3 protein is expressed in the Purkinje cells of the cerebellum, again a proof of a neuronal D3 expression (Verhoelst et al. 2002). However, the present study reports the presence of D2 in a non-neuronal cell type, namely the epithelial cells of the choroid plexus. If the avian blood–brain barrier mechanism is similar to that of mammals, the high expression of the D2 protein in the choroidal epithelial cells seems rather surprising. It has been proven, however, that chicken TTR shows little affinity for T₄, while it binds T₃ very well (Chang et al. 1999). This would explain why the D2 protein is indeed present in the choroid plexus. The D2 protein deiodinates T₄ that enters the choroid plexus via the basolateral membrane in order to form T₃, which can subsequently bind to TTR to allow further transport through the cerebrospinal fluid (Fig. 3). This would suggest that astrocytes are not the sole source for active neuronal TH and that both the choroid plexus and the astroglial cells might be responsible for the T₃ flow towards the neurons. Further support for this similar function of the astrocytes and the choroidal epithelial cells comes from their developmental origin. As mentioned before, the epithelial cells of the choroid plexus are generated by fusion of the ependymal cells lining the ventricles and the pia mater. The ependymal cells lining the ventricular cavities, however, do not solely give rise to the choroidal epithelial cells but partially differentiate to form glial cells, like astrocytes, as well. This common origin of both types of cells may explain their similar TH-activating task. Furthermore, the immunohistochemical stainings performed in this study showed the staining of the cells migrating from the lining of the lateral ventricle, indicating that D2 protein is already present in these cells. Therefore, the authors propose this new hypothesis concerning the transport of thyroid hormones in the cerebrospinal fluid in chicken. A schedule based on the earlier published mammalian blood–brain barrier mechanism is introduced. Whether the expression of D2 protein in the embryonic chicken choroid plexus is a temporary, and thus a transient development related expression, should be elucidated further.

In conclusion, the present study reports, for the first time, the expression of D2 in the cuboidal epithelial cells of the choroid plexus. No expression of other deiodinase proteins could be found at the level of this blood–brain barrier. Dexamethasone treatment led to an up-regulation of the D2 protein in the cuboidal cells of the choroid plexus. This is the first evidence of the D2 enzyme in this vertebrate blood–brain barrier structure so far. The presence of D2 protein in the choroid plexus, along with the expression of D2 enzyme along the lining of the ventricular walls, leads to the proposal of a novel mechanism concerning the transport of thyroid hormones along the blood-brain barrier in birds in comparison with the previously established working mechanism for mammalian TH transport.

	Acknowledgements

 
We wish to thank Willy Van Ham, Francine Voets, Lut Noterdaeme and Tina Everaert for their valuable technical assistance. Furthermore, we wish to acknowledge L Arckens, E Van der Gucht and S Jacobs for their help with image analysis. This work was financed by the KU Leuven Research Council Grant OT-01–23, Fonds voor Wetenschappelijk Onderzoek Vlaanderen Grant G.O272.04 and European Union Grant QLG3-CT2000–00930. Serge Van der Geyten was supported by the Fund for Scientific Research – Flanders.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 
Bernal J, Guadaño-Ferraz A & Morte B 2003 Perspectives in the study of thyroid hormone action on brain development and function. Thyroid 13 1005–1012.[CrossRef][ISI][Medline]

Bianco AC, Salvatore D, Gereben B, Berry MJ & Larsen PR 2002 Biochemistry, cellular and molecular biology, and physiological roles of the iodothyronine selenodeiodinases. Endocrine Reviews 23 38–89.[Abstract/Free Full Text]

Callebaut I, Curcio-Morelli C, Mornon JP, Gereben B, Buettner C, Huang S, Castro B, Fonseca TL, Harney JW, Larsen PR & Bianco AC 2003 The iodothyronine selenodeiodinases are thioredoxin-fold family proteins containing a glycoside hydrolase clan GH-A-like structure. Journal of Biological Chemistry 278 36687–36696.

Chang L, Munro SLA, Richardson SJ & Schreiber G 1999 Evolution of thyroid binding in transthyretins in birds and mammals. European Journal of Biochemistry 259 534–542.[ISI][Medline]

Diano S, Leonard JL, Meli R, Esposito E & Schiavo L 2003 Hypothalamic type II iodothyronine deiodinase: a light and electron microscopic study. Brain Research 976 130–134.[CrossRef][ISI][Medline]

Friesema ECH, Ganguly S, Abdalla A, Manning Fox JE, Halestrap AP & Visser TJ 2003 Identification of monocarboxylate transporter 8 as a specific thyroid hormone transporter. Journal of Biological Chemistry 278 40128–40135.[Abstract/Free Full Text]

Gereben B, Bartha T, Tu HM, Rudas P & Larsen PR 1999 Cloning and expression of the chicken type II iodothyronine 5'-deiodinase. Journal of Biological Chemistry 274 13768–13776.[Abstract/Free Full Text]

Gereben B, Pachucki J, Kollár A, Liposits Z & Fekete C 2004 Ontogenetic redistribution of type 2 iodothyronine mRNA in the brain of chicken. Endocrinology 145 3619–3625.[Abstract/Free Full Text]

Guadaño-Ferraz A, Obregón MJ, St Germain DL & Bernal J 1997 The type 2 iodothyronine deiodinase is expressed primarily in glial cells in the neonatal rat brain. PNAS 94 10391–10396.[Abstract/Free Full Text]

Guadaño-Ferraz A, Escaméz MJ, Raussell E & Bernal J 1999 Expression of type 2 iodothyronine deiodinase in hypothyroid rat brain indicates an important role of thyroid hormone in the development of specific primary sensory systems. Journal of Neuroscience 19 3430–3439.[Abstract/Free Full Text]

Leonard JL & Visser TJ 1986 Biochemistry of deiodination. In Thyroid Hormone Metabolism, pp 189–229. Ed G Hennemann. New York, USA: Marcel Dekker.

Tu HM, Kim S-W, Salvatore D, Bartha T, Legradi G, Larsen PR & Lechan RM 1997 Regional distribution of type 2 thyroxine deiodinase messenger ribonucleic acid in rat hypothalamus and pituitary and its regulation by thyroid hormone. Endocrinology 138 3359–3368.[Abstract/Free Full Text]

Schreiber G 2002 The evolution of transthyretin synthesis in the choroid plexus. Clinical and Chemical Laboratory Medicine 40 1200–1210.[CrossRef][ISI][Medline]

Simpson IA 2004. URL: http://www.hmc.psu.edu/photos/researchgraphics/isimpson.jpg.

Southwell BR, Duan W, Alcorn D, Brack C, Richardson SJ, Köhrle J & Schreiber G 1993 Thyroxine transport to the brain: Role of protein synthesis by the choroid plexus. Endocrinology 133 2116–2126.[Abstract]

Speake T, Whitwell C, Kajita H, Majid A & Brown PD 2001 Mechanisms of CSF secretion by the choroid plexus. Microscopy Research Techniques 52 49–59.

Van der Geyten S, Segers I, Gereben B, Bartha T, Rudas P, Larsen PR, Kühn ER & Darras VM 2001 Transcriptional regulation of iodothyronine deiodinases during embryonic development. Molecular and Cellular Endocrinology 183 1–9.[CrossRef][ISI][Medline]

Verhoelst CHJ, Vandenborne K, Severi T, Bakker O, Zandieh Doulabi B, Leonard JL, Van der Geyten S, Kühn ER & Darras VM 2002 Specific detection of type III iodothyronine deiodinase protein in chicken cerebellum. Endocrinology 143 2700–2707.[Abstract/Free Full Text]

Verhoelst CHJ, Darras VM, Zandieh Doulabi B, Reyns GE, Kühn ER & Van der Geyten S 2003 Type I iodothyronine deiodinase in euthyroid and hypothyroid chicken cerebellum. Molecular and Celular Endocrinology 214 97–105.

Received 13 May 2004
Accepted 14 July 2004
Made available online as an Accepted Preprint 27 July 2004

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