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Measurement of messenger RNA encoding the α-chain, polymeric immunoglobulin receptor, and J-chain in duodenal mucosa from dogs with and without chronic diarrhea by use of quantitative real-time reverse transcription-polymerase chain reaction assays

Iain R. PetersSchool of Clinical Veterinary Science, University of Bristol, Langford, Bristol, BS40 5DU, UK.

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 BVMS (Hons), PhD
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Chris R. HelpsSchool of Clinical Veterinary Science, University of Bristol, Langford, Bristol, BS40 5DU, UK.

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 PhD
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Emma L. CalvertWALTHAM Centre for Pet Nutrition, Waltham on the Wolds, Leicestershire, LE14 4RT, UK.

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 PhD
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Edward J. HallSchool of Clinical Veterinary Science, University of Bristol, Langford, Bristol, BS40 5DU, UK.

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 VetMB, PhD
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Michael J. DaySchool of Clinical Veterinary Science, University of Bristol, Langford, Bristol, BS40 5DU, UK.

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 BVMS (Hons), PhD
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Abstract

Objective—To examine the difference in expression of messenger RNA (mRNA) transcripts for polymeric immunoglobulin receptor (pIgR), α-chain, and J-chain determined by use of quantitative real-time reverse transcription-polymerase chain reaction (QRT-PCR) assays in duodenal biopsy specimens obtained from dogs with and without chronic diarrhea.

Sample Population—Biopsy specimens of the proximal portion of the duodenum were obtained endoscopically from 39 dogs evaluated because of chronic diarrhea (12 German Shepherd Dogs and 27 non-German Shepherd Dog breeds); specimens were also obtained from a control group of 7 dogs evaluated because of other gastrointestinal tract diseases and 2 dogs that were euthanatized as a result of nongastrointestinal tract disease.

Procedure—Dogs were anesthetized, and multiple mucosal biopsy specimens were obtained endoscopically at the level of the caudal duodenal flexure by use of biopsy forceps; in 2 control dogs, samples were obtained from the descending duodenum within 5 minutes of euthanasia. One-step QRT-PCR was used to quantify the level of expression of transcripts for the housekeeper gene glyceraldehyde-3-phosphate dehydrogenase, pIgR, α-chain, and J-chain in duodenal mucosal tissue.

Results—There was no significant difference in the level of expression of any transcript among non-German Shepherd Dog breeds without diarrhea (control group), non-German Shepherd Dog breeds with chronic diarrhea, and German Shepherd Dogs with chronic diarrhea.

Conclusions and Clinical Relevance—Results indicated that the susceptibility of German Shepherd Dogs to chronic diarrhea is not a result of simple failure of transcription of the key genes that encode molecules involved in mucosal IgA secretion. (Am J Vet Res 2005;66:11–16)

Abstract

Objective—To examine the difference in expression of messenger RNA (mRNA) transcripts for polymeric immunoglobulin receptor (pIgR), α-chain, and J-chain determined by use of quantitative real-time reverse transcription-polymerase chain reaction (QRT-PCR) assays in duodenal biopsy specimens obtained from dogs with and without chronic diarrhea.

Sample Population—Biopsy specimens of the proximal portion of the duodenum were obtained endoscopically from 39 dogs evaluated because of chronic diarrhea (12 German Shepherd Dogs and 27 non-German Shepherd Dog breeds); specimens were also obtained from a control group of 7 dogs evaluated because of other gastrointestinal tract diseases and 2 dogs that were euthanatized as a result of nongastrointestinal tract disease.

Procedure—Dogs were anesthetized, and multiple mucosal biopsy specimens were obtained endoscopically at the level of the caudal duodenal flexure by use of biopsy forceps; in 2 control dogs, samples were obtained from the descending duodenum within 5 minutes of euthanasia. One-step QRT-PCR was used to quantify the level of expression of transcripts for the housekeeper gene glyceraldehyde-3-phosphate dehydrogenase, pIgR, α-chain, and J-chain in duodenal mucosal tissue.

Results—There was no significant difference in the level of expression of any transcript among non-German Shepherd Dog breeds without diarrhea (control group), non-German Shepherd Dog breeds with chronic diarrhea, and German Shepherd Dogs with chronic diarrhea.

Conclusions and Clinical Relevance—Results indicated that the susceptibility of German Shepherd Dogs to chronic diarrhea is not a result of simple failure of transcription of the key genes that encode molecules involved in mucosal IgA secretion. (Am J Vet Res 2005;66:11–16)