Molecular Cloning, Characterization And Expression Of A Novel Retinal Clusterin-Like Protein Cdna, Qi Zhang, Kunal Ray, Gregory M. Acland, Jill M. Czarnecki, Gustavo D. Aguirre
Jan 2000
Molecular Cloning, Characterization And Expression Of A Novel Retinal Clusterin-Like Protein Cdna, Qi Zhang, Kunal Ray, Gregory M. Acland, Jill M. Czarnecki, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
A novel gene expressed predominantly in retina, but detected at a conspicuously lower level in retina of canine progressive rod cone degeneration (prcd), has been identified by suppression subtractive hybridization and retinal cDNA library screening. The characterized region of cDNA of the novel gene includes 1017 nucleotides of coding sequence predicted to encode a protein of 338 amino acids (Mr 39 389), 791 nucleotides of 5′-untranslated region (UTR), and 300 nucleotides of 3′-UTR including the poly(A)+ tail. Multiple transcripts were detected in retina by Northern blot analysis, and a lower level of expression was observed in …
Evaluation Of Cgmp-Phosphodiesterase (Pde) Subunits For Causal Association With Rod–Cone Dysplasia 2 (Rcd2), A Canine Model Of Abnormal Retinal Cgmp Metabolism, Weiquan Wang, Gregory Acland, Kunal Ray, Gustavo Aguirre
Sep 1999
Evaluation Of Cgmp-Phosphodiesterase (Pde) Subunits For Causal Association With Rod–Cone Dysplasia 2 (Rcd2), A Canine Model Of Abnormal Retinal Cgmp Metabolism, Weiquan Wang, Gregory Acland, Kunal Ray, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
Rod-cone dysplasia types 1 (rcd1; Irish setter) and 2 (rcd2; collie) in dogs are early onset forms of progressive retinal atrophy (PRA) which serve as models of retinitis pigmentosa(RP) in humans. As bothrcd1 and rcd2 result from abnormal retinal cGMP metabolism associated with a deficiency in cGMP-phosphodiesterase (PDE) activity, and a nonsense mutation in the PDE6B subunit gene has been shown to cause rcd1, the genes encoding the four subunits of the PDE complex (PDE6A, PDE6B, PDE6G and PDE6D) make compelling candidates for the rcd2 locus. We adopted diverse strategies to evaluate causal association of the four PDE subunit genes …
Synthesis And Release Of Docosahexaenoic Acid By The Rpe Cells Of Prcd-Affected Dogs, Huiming Chen, Jharna Ray, Virginia Scarpino, Gregory M. Acland, Gustavo D. Aguirre, Robert E. Anderson
Aug 1999
Synthesis And Release Of Docosahexaenoic Acid By The Rpe Cells Of Prcd-Affected Dogs, Huiming Chen, Jharna Ray, Virginia Scarpino, Gregory M. Acland, Gustavo D. Aguirre, Robert E. Anderson
Gustavo D. Aguirre, VMD, PhD
Purpose: Dogs affected with progressive rod-cone degeneration (prcd) have reduced levels of docosahexaenoic acid (DHA, 22:6n-3) in their plasma and rod photoreceptor outer segments (ROS). Dietary supplementation of DHA has failed to increase the ROS DHA levels to that of unaffected control dogs. The present study was undertaken to test the hypothesis that prcd-affected dogs have a reduced capacity for the synthesis and/or release of DHA in retinal pigment epithelial (RPE) cells.
Methods: RPE cells (first passage cultures) from prcd-affected and normal dogs were incubated with [3H]eicosapentaenoic acid (EPA, 20:5n-3) for 24 and 72 hours. After …
A Novel Retinal Degeneration Locus Identified By Linkage And Comparative Mapping Of Canine Early Retinal Degeneration, Gregory Acland, Kunal Ray, Cathryn Mellersh, Amelia Langston, Jasper Rine, Elaine Ostrander, Gustavo Aguirre
Jul 1999
A Novel Retinal Degeneration Locus Identified By Linkage And Comparative Mapping Of Canine Early Retinal Degeneration, Gregory Acland, Kunal Ray, Cathryn Mellersh, Amelia Langston, Jasper Rine, Elaine Ostrander, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
Early retinal degeneration (erd) is an early onset progressive retinal atrophy, a hereditary canine retinal disease phenotypically similar to human retinitis pigmentosa (RP). In previous efforts to identify the erd locus, canine homologs of genes causally associated with RP in humans, such as opsin (RHO), the β-subunit gene for cyclic GMP phosphodiesterase (PDE6B), and RDS/peripherin, were excluded. A genome-wide screen was undertaken on canine families segregating the erd disease. Analysis of over 150 canine-specific markers has localized erd to a single linkage group comprising two previously identified canine linkage groups, 20 and 26, corresponding to canine radiation hybrid groups RH.34-a …
Photoreceptor Dysplasia (Pd) In Miniature Schnauzer Dogs: Evaluation Of Candidate Genes By Molecular Genetic Analysis, Qi Zhang, V J. Baldwin, Gregory M. Acland, C J. Parshall, J. Haskell, Gustavo D. Aguirre, Kunal Ray
Dec 1998
Photoreceptor Dysplasia (Pd) In Miniature Schnauzer Dogs: Evaluation Of Candidate Genes By Molecular Genetic Analysis, Qi Zhang, V J. Baldwin, Gregory M. Acland, C J. Parshall, J. Haskell, Gustavo D. Aguirre, Kunal Ray
Gustavo D. Aguirre, VMD, PhD
Photoreceptor dysplasia (pd) is one of a group of at least six distinct autosomal and one X-linked retinal disorders identified in dogs which are collectively known as progressive retinal atrophy (PRA). It is an early onset retinal disease identified in miniature schnauzer dogs, and pedigree analysis and breeding studies have established autosomal recessive inheritance of the disease. Using a gene-based approach, a number of retina-expressed genes, including some members of the phototransduction pathway, have been causally implicated in retinal diseases of humans and other animals. Here we examined seven such potential candidate genes (opsin, RDS/peripherin, ROM1, rod cGMP-gated cation channel …
Overview Of The International Workshop On Canine Genetics, Gustavo D. Aguirre, Kunal Ray, Gregory M. Acland
Dec 1998
Overview Of The International Workshop On Canine Genetics, Gustavo D. Aguirre, Kunal Ray, Gregory M. Acland
Gustavo D. Aguirre, VMD, PhD
No abstract provided.
Timp-1 Expression Is Increased In X-Linked Progressive Retinal Atrophy Despite Its Exclusion As A Candidate Gene, Caroline Zeiss, Gregory M. Acland, Gustavo D. Aguirre, Kunal Ray
Dec 1998
Timp-1 Expression Is Increased In X-Linked Progressive Retinal Atrophy Despite Its Exclusion As A Candidate Gene, Caroline Zeiss, Gregory M. Acland, Gustavo D. Aguirre, Kunal Ray
Gustavo D. Aguirre, VMD, PhD
X-linked progressive retinal atrophy (XLPRA) is the only known natural animal model for X-linked retinitis pigmentosa (XLRP), a blinding disorder in man. The tissue inhibitor metalloproteinase 1 gene (TIMP-1), present in close proximity to one of the two XLRPloci, was tested as a candidate for XLPRA, by first characterizing the cDNA and gene from a normal dog. The cloned canine TIMP-1 cDNA is predicted to encode a protein of 207 amino acids with 66–83% identity in the deduced aa sequence with homologous mammalian genes. No sequence difference in the coding sequence of …
Canine Cone Transducin-Gamma Gene And Cone Degeneration In The Cd Dog, Gustavo D. Aguirre
Aug 1998
Canine Cone Transducin-Gamma Gene And Cone Degeneration In The Cd Dog, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Purpose: To characterize the cDNA and the organization of the gene encoding the cone-specific gamma subunit of transducin (Tgamma c) and to examine this gene as a candidate for the recessively inherited cone photoreceptor degeneration in the cd dog.
Methods: Canine Tgamma c cDNA was cloned and sequenced. Polymerase chain reaction (PCR) was used to define the Tgamma c gene structure, northern blot analysis to examine the level of expression of Tgamma c mRNA in control and cd-affected retinas, and immunocytochemistry to determine the presence and localization of Tgamma c in normal and cd retinas.
Results: Immunocytochemical results showed Tgamma …
Identification Of A Rapd Marker Linked To Progressive Rod-Cone Degeneration In Dogs, Weikuan Gu, Gregory M. Acland, Amelia A. Langston, Elaine A. Ostrander, Gustavo D. Aguirre, Kunal Ray
Aug 1998
Identification Of A Rapd Marker Linked To Progressive Rod-Cone Degeneration In Dogs, Weikuan Gu, Gregory M. Acland, Amelia A. Langston, Elaine A. Ostrander, Gustavo D. Aguirre, Kunal Ray
Gustavo D. Aguirre, VMD, PhD
Random amplified polymorphic DNA (RAPD) analysis has been used widely in plant and fungi for identification of markers linked to genetic traits and mapping, but its use is limited to identification of intra- and inter-species difference in domestic mammals. We report here identification of a RAPD-derived marker linked to progressive rod-cone degeneration (prcd), an inherited autosomal recessive retinal disease of dogs. A total of 400 standard 10-mer primers were used for amplification by use of DNA samples from normal (+/+) and affected (prcd/prcd) dogs. A single primer was identified which amplified a 1.5-kb DNA fragment only from normal dogs. PCR …
Characterization Of Canine Photoreceptor Phosducin Cdna And Identification Of A Sequence Variant In Dogs With Photoreceptor Dysplasia, Qi Zhang, Gregory M. Acland, Charles J. Parhsall, Jeanette Haskell, Kunal Ray, Gustavo D. Aguirre
Jul 1998
Characterization Of Canine Photoreceptor Phosducin Cdna And Identification Of A Sequence Variant In Dogs With Photoreceptor Dysplasia, Qi Zhang, Gregory M. Acland, Charles J. Parhsall, Jeanette Haskell, Kunal Ray, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Photoreceptor dysplasia (pd) is an autosomal recessive disease of miniature schnauzer dogs causing retinal degeneration. The disease is a homologue of retinitis pigmentosa, a group of genetically heterogeneous diseases, causing blindness in humans. A subtraction library was prepared from retinas of pd affected and age-matched normal control dogs to isolate de novo candidate genes for further examination. From the subtraction library, cDNA for phosducin (PDC), a member of the phototransduction pathway, was isolated as a transcript expressed at a higher level in the affected retina. First, the normal canine PDC cDNA was characterized to evaluate the PDC gene …
Cloning Of The Canine Β-Glucuronidase Cdna, Mutation Identification In Canine Mps Vii, And Retroviral Vector-Mediated Correction Of Mps Vii Cells, Jharna Ray, Alain Bouvet, Christopher Desanto, John Fyfe, Danbin Xu, John Wolfe, Gustavo Aguirre, Donald Patterson, Mark Haskins, Paula Henthorn
Feb 1998
Cloning Of The Canine Β-Glucuronidase Cdna, Mutation Identification In Canine Mps Vii, And Retroviral Vector-Mediated Correction Of Mps Vii Cells, Jharna Ray, Alain Bouvet, Christopher Desanto, John Fyfe, Danbin Xu, John Wolfe, Gustavo Aguirre, Donald Patterson, Mark Haskins, Paula Henthorn
Gustavo D. Aguirre, VMD, PhD
Mucopolysaccharidosis type VII (MPS VII) is an inherited disease resulting from deficient activity of the lysosomal acid hydrolase β-glucuronidase (GUSB) and has been reported in humans, mice, cats, and dogs. To characterize canine MPS VII, we have isolated and sequenced the canine GUSB cDNA from normal and affected animals. A single nucleotide substitution was detected in the GUSB cDNA derived from MPS VII dogs. This guanosine to adenine base change at nucleotide position 559 in the canine cDNA sequence causes an arginine to histidine substitution at amino acid position 166. Introduction of the G to A substitution at position 559 …
A Linkage Map Of The Canine Genome, Cathryn Mellersh, Amelia Langston, Gregory Acland, Melissa Fleming, Kunal Ray, Neil Weigand, Leigh Francisco, Mark Gibbs, Gustavo Aguirre, Elaine Ostrander
Dec 1997
A Linkage Map Of The Canine Genome, Cathryn Mellersh, Amelia Langston, Gregory Acland, Melissa Fleming, Kunal Ray, Neil Weigand, Leigh Francisco, Mark Gibbs, Gustavo Aguirre, Elaine Ostrander
Gustavo D. Aguirre, VMD, PhD
A genetic linkage map of the canine genome has been developed by typing 150 microsatellite markers using 17 three-generation pedigrees, composed of 163 F2individuals. One hundred and thirty-nine markers were linked to at least one other marker with a lod score ≥ 3.0, identifying 30 linkage groups. The largest chromosome had 9 markers spanning 106.1 cM. The average distance between markers was 14.03 cM, and the map covers an estimated 2073 cM. Eleven markers were informative on the mapping panel, but were unlinked to any other marker. These likely represent single markers located on small, distinct canine chromosomes. This map …
Construction Of A Panel Of Canine–Rodent Hybrid Cell Lines For Use In Partitioning Of The Canine Genome, Amelia A. Langston, Cathryn S. Mellersh, Cassandra L. Neal, Kunal Ray, Gregory M. Acland, Mark Gibbs, Gustavo D. Aguirre, R.E. K. Fournier, Elaine A. Ostrander
Dec 1997
Construction Of A Panel Of Canine–Rodent Hybrid Cell Lines For Use In Partitioning Of The Canine Genome, Amelia A. Langston, Cathryn S. Mellersh, Cassandra L. Neal, Kunal Ray, Gregory M. Acland, Mark Gibbs, Gustavo D. Aguirre, R.E. K. Fournier, Elaine A. Ostrander
Gustavo D. Aguirre, VMD, PhD
We have constructed a collection of canine–rodent microcell hybrid cell lines by fusion of canine fibroblast microcell donors with immortalized rodent recipient cells. Characterization of the hybrid cell lines using a combination of fluorescencein situhybridization and PCR analysis of canine microsatellite repeat sequences allowed selection of a panel of hybrids in which most canine chromosomes are represented. Approximately 90% of genetic markers and genes that were tested could be assigned to 1 of 31 anonymous canine chromosome groups, based on common patterns of retention in the hybrid set. Many of these putative chromosome groups have now been validated …
Canine Rod Photoreceptor Cgmp-Gated Channel Protein Α-Subunit: Studies On The Expression Of The Gene And Characterization Of The Cdna, Qi Zhang, Sue Pearce-Kelling, Gregory Acland, Gustavo Aguirre, Kunal Ray
Jul 1997
Canine Rod Photoreceptor Cgmp-Gated Channel Protein Α-Subunit: Studies On The Expression Of The Gene And Characterization Of The Cdna, Qi Zhang, Sue Pearce-Kelling, Gregory Acland, Gustavo Aguirre, Kunal Ray
Gustavo D. Aguirre, VMD, PhD
Rod photoreceptor cyclic GMP gated-channel protein is a key component of the visual transduction cascade in the vertebrate retina. The protein is composed of at least two subunits (α and β). Mutations in the α-subunit (CNGC1) have been shown to cause retinitis pigmentosa (RP) in humans. Several heterogeneous canine retinal diseases, which are clinically similar to RP, are known collectively as progressive retinal atrophy (PRA) and occur in dogs in a breed-specific manner. For the purpose of examining CNGC1 gene as a candidate for PRA, we report here the characterization of canine CNGC1 cDNA, and examine the expression of the …
Differential Expression Of Photoreceptor-Specific Proteins During Disease And Degeneration In The Progressive Rod-Cone Degeneration (Prcd) Retina, Kathryn Gropp, Jun Huang, Gustavo Aguirre
May 1997
Differential Expression Of Photoreceptor-Specific Proteins During Disease And Degeneration In The Progressive Rod-Cone Degeneration (Prcd) Retina, Kathryn Gropp, Jun Huang, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
Progressive rod-cone degeneration (prcd) is a late-onset hereditary retinal degeneration characterized by normal development of photoreceptors prior to degeneration and death of visual cells. We reported previously that expression of opsin mRNA and protein decreases prior to visual cell degeneration. To examine the specificity of this reduction, we have used immunocytochemistry to correlate photoreceptor-specific protein expression with visual cell disease progression. Eyes from light-adapted age-matched control andprcd-affected dogs were fixed in paraformaldehyde, embedded in diethylene glycol distearate (DGD) wax, and reacted with antibodies specific to interphotoreceptor retinoid-binding protein (IRBP), S-antigen, opsin, phosducin, γ-phosphodiesterase (γ-PDE), and β1-transducin. While IRBP expression did …
Canine Rod Transducin A-1: Cloning Of The Cdna And Evaluation Of The Gene As A Candidate For Progressive Retinal Atrophy, Kunal Ray, Victoria J. Baldwin, Caroline Zeiss, Gregory M. Acland, Gustavo D. Aguirre
Dec 1996
Canine Rod Transducin A-1: Cloning Of The Cdna And Evaluation Of The Gene As A Candidate For Progressive Retinal Atrophy, Kunal Ray, Victoria J. Baldwin, Caroline Zeiss, Gregory M. Acland, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Purpose: Progressive retinal atrophy (PRA) represents a heterogeneous group of retinal dystrophies, distinct forms of which occur in different canine breeds. The present study was undertaken to evaluate the gene for the a-1 subunit of the rod specific G-protein transducin (GNAT1), a member of the phototransduction pathway, as a candidate for progressive rod cone degeneration (prcd) in poodles, early retinal degeneration (erd) in elkhounds, and rod cone dysplasia 2 (rcd 2) in collies.
Methods: Oligonucleotide primers were designed from the consensus region of known cDNA sequences for GNAT1 from other species. Canine GNAT1 cDNA was cloned and sequenced after reverse …
Selective Absence Of Cone Outer Segment Β3-Transducin Immunoreactivity In Hereditary Cone Degeneration (Cd), Kathryn Gropp, A. Széll, Jun Huang, Gregory Acland, Debora Farber, Gustavo Aguirre
Aug 1996
Selective Absence Of Cone Outer Segment Β3-Transducin Immunoreactivity In Hereditary Cone Degeneration (Cd), Kathryn Gropp, A. Széll, Jun Huang, Gregory Acland, Debora Farber, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In thecdretina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later, by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ …
Nonallelism Of Erd And Prcd And Exclusion Of The Canine Rds/Peripherin Gene As A Candidate For Both Retinal Degeneration Loci., K Ray, Gregory M. Acland, Gustavo D. Aguirre
Mar 1996
Nonallelism Of Erd And Prcd And Exclusion Of The Canine Rds/Peripherin Gene As A Candidate For Both Retinal Degeneration Loci., K Ray, Gregory M. Acland, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Purpose: To determine whether early retinal degeneration (erd) and progressive rod cone degeneration (prcd), two canine hereditary retinal degenerations, are caused by allelic mutations; to determine the cDNA sequence of the canine RDS/peripherin homolog (CFRDSP); and to test whether mutations(s) in CFRDSP cause(s) either erd or prcd.
Methods: Three erd-affected dogs were crossbred to three prcd-affected dogs, and their progeny were tested by electroretinography and retinal morphology for evidence of retinal degeneration. Canine RDS/peripherin cDNA was cloned and sequenced after reverse-transcription-polymerase chain reaction (RT-PCR) of total retinal RNA. A set of overlapping fragments of CFRDSP cDNA amplified from normal and …
Redistribution Of Insoluble Interphotoreceptor Matrix Components During Photoreceptor Differentiation In The Mouse Retina, Kristina Mieziewska, Agoston Szel, Theo Van Veen, Gustavo D. Aguirre, Nancy Philp
Jun 1994
Redistribution Of Insoluble Interphotoreceptor Matrix Components During Photoreceptor Differentiation In The Mouse Retina, Kristina Mieziewska, Agoston Szel, Theo Van Veen, Gustavo D. Aguirre, Nancy Philp
Gustavo D. Aguirre, VMD, PhD
The development of the nervous-system is largely influenced by the extracellular matrix (ECM). In the neural retina, the photoreceptors are surrounded by a unique ECM, the interphotoreceptor matrix (IPM). The IPM plays a central and possibly crucial role in the development, maintenance and specific function of the photoreceptors. Therefore, the characterization of IPM components is necessary to understand the mechanisms regulating photoreceptor differentiation.
The IPM in the mouse retina was examined during photoreceptor morphogenesis with the monoclonal antibody (MAb) F22, which recognizes a 250 kDa component of the interphotoreceptor matrix. The binding pattern of MAb F22 revealed a striking redistribution …
Plasma Lipid Changes In Prcd-Affected And Normal Miniature Poodles Given Oral Supplements Of Linseed Oil. Indications For The Involvement Of N-3 Fatty Acids In Inherited Retinal Degenerations, Robert E. Anderson, Maureen B. Maude, Gregory M. Acland, Gustavo D. Aguirre
Jan 1994
Plasma Lipid Changes In Prcd-Affected And Normal Miniature Poodles Given Oral Supplements Of Linseed Oil. Indications For The Involvement Of N-3 Fatty Acids In Inherited Retinal Degenerations, Robert E. Anderson, Maureen B. Maude, Gregory M. Acland, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
It has previously been shown that miniature poodles with regressive rod-cone degeneration (PRCD) have lower plasma levels of docosahexaenoic acid (22:6n-3) than normal poodles and it has been suggested that affected animals have a defect in the motabolism of 22:6n-3. To test this hypothesis in vivo, PRCD affected and normal miniature poodles were given daily oral supplements of linsee oil (enriched in 19:3n-3, 20:5n-3, and 22:5n-3 between affected and normal dogs. Therefore, there appears to be no abnormality in the elongation and desaturation system that takes 18:3n-3 and 22:5n-3. Surprisingly, the plasma level of 22:6n-3 was reduced in both groups …
Decreased Opsin Mrna And Immunoreactivity In Progressive Rod-Cone Degeneration (Prcd): Cytochemical Studies Of Early Disease And Degeneration, Jun Huang, Marie-Françoise Chesselet, Gustavo Aguirre
Dec 1993
Decreased Opsin Mrna And Immunoreactivity In Progressive Rod-Cone Degeneration (Prcd): Cytochemical Studies Of Early Disease And Degeneration, Jun Huang, Marie-Françoise Chesselet, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
Opsin mRNA level and immunoreactivity were examined by in situ hybridization and immunocytochemistry in normal and progressive rod cone degeneration (prcd)-affected dogs. In situ hybridization used 35S- and/or 3 H-labeled bovine opsin cRNA probes; immunocytochemistry used six monoclonal mouse anti-bovine opsin antibodies (MAb1) that are specific to different regions of the N-terminal, loop v-vi and the C-terminal domains. Optimal labeling and histological resolution at the single cell level were achieved with semi-thin sections of DGD wax-embedded tissues; it was possible to correlate the cytochemical observations with the disease staging in topographically defined regions that exhibited different disease severity. In early …
Diethylene Glycol Distearate (Dgd): A Versatile Embedding Medium For Retinal Cytochemistry, Jun C. Huang, Kristina Mieziewska, Nancy Philp, Theo Van Veen, Gustavo D. Aguirre
Apr 1993
Diethylene Glycol Distearate (Dgd): A Versatile Embedding Medium For Retinal Cytochemistry, Jun C. Huang, Kristina Mieziewska, Nancy Philp, Theo Van Veen, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Embedment in diethylene glycol distearate (DGD) was shown to be highly desirable and versatile for retinal cytochemical studies, including in situ hybridization, immuno- andlectin cytochemistry. This method allows for preservation of fine tissue detail as well as good reaction sensitivity. It appears to be more suitable than most other methods currently used for light microscopic retinal cytochemistry.
The Cone Matrix Sheath In The Normal And Diseased Retina: Cytochemical And Biochemical Studies Of Peanut Agglutinin-Binding Proteins In Cone And Rod-Cone Degeneration, Kenneth Long, Gustavo Aguirre
May 1991
The Cone Matrix Sheath In The Normal And Diseased Retina: Cytochemical And Biochemical Studies Of Peanut Agglutinin-Binding Proteins In Cone And Rod-Cone Degeneration, Kenneth Long, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
The fate of the cone-associated extracellular domain, or cone matrix sheath (CMS), was examined in two canine models of hereditary retinal degeneration. The diseases, which affect cones selectively (cd = cone degeneration), or rods and cones temporally (prcd = progressive rod-cone degeneration), were examined biochemically (SDS-PAGE/lectinblots) and cytochemically (light microscopy) using peanut agglutinin lectin (PNA) to selectively label this domain and associated structures. Most of the cones had disappeared in the adult cd retina. In the remaining cones, PNA labeled the ectopically located somata and the CMSs that were present around severely diseased ones. …
Glycosaminoglycan And Collagen Metabolism In Arylsulfatase B-Deficient Retinal Pigment Epithelium In Vitro, Gustavo D. Aguirre, Lawrence E. Stramm, W Li, M Haskins
May 1991
Glycosaminoglycan And Collagen Metabolism In Arylsulfatase B-Deficient Retinal Pigment Epithelium In Vitro, Gustavo D. Aguirre, Lawrence E. Stramm, W Li, M Haskins
Gustavo D. Aguirre, VMD, PhD
Regional differences in retinal pigment epithelial (RPE) cell glycosaminoglycan (GAG) and collagen metabolism were studied using cells obtained from normal cats and those with deficient activity of arylsulfatase B (ASB), a lysosomal enzyme involved in GAG catabolism. Control and ASB-deficient RPE cultures initiated from superior equatorial (superior) and inferior equatorial (inferior) regions of the eye were radiolabeled for 72 hr with 35SO4, and GAGs from the media and cell layers were analyzed separately. In ASB-deficient RPE, there was an accumulation of dermatan/chondroitin sulfate in the cell layer of cultures initiated from the superior region of the eye but not in …
Β-Glucuronidase Mediated Pathway Essential For Retinal Pigment Epithelial Degradation Of Glycosaminoglycans. Disease Expression And In Vitro Disease Correction Using Retroviral Mediated Cdna Transfer, Lawrence Stramm, John Wolfe, Edward Schuchman, Mark Haskins, Donald Patterson, Gustavo Aguirre
Apr 1990
Β-Glucuronidase Mediated Pathway Essential For Retinal Pigment Epithelial Degradation Of Glycosaminoglycans. Disease Expression And In Vitro Disease Correction Using Retroviral Mediated Cdna Transfer, Lawrence Stramm, John Wolfe, Edward Schuchman, Mark Haskins, Donald Patterson, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
A β-glucuronidase mediated pathway for the degradation of glycosaminoglycans is present in the retinal pigment epithelium. The pathway has been defined using ocular tissues and cultured cells from mutant animals having a recessively inherited deficiency of the lysosomal enzyme. In situ, storage products accumulate in secondary lysosomes of the retinal pigment epithelium, the cytoplasm fills with inclusions and the cells hypertrophy; severity of the disease increases with aging. Deficient activity of β-glucuronidase is present in primary and second passage cultures. Radiolabel studies with 35SO4 show a significant retention of cell layer label by mutant retinal pigment epithelial cells …
Segregation Distortion In Inheritance Of Progressive Rod Cone Degeneration (Prcd) In Miniature Poodle Dogs, Gregory M. Acland, Susan Halloran-Blanton, Joann A. Boughman, Gustavo D. Aguirre
Feb 1990
Segregation Distortion In Inheritance Of Progressive Rod Cone Degeneration (Prcd) In Miniature Poodle Dogs, Gregory M. Acland, Susan Halloran-Blanton, Joann A. Boughman, Gustavo D. Aguirre
Gustavo D. Aguirre, VMD, PhD
Segregation distortion was observed in inheritance of progressive rod–cone degeneration (prcd) in a colony of Miniature Poodle dogs. Breeding results, from both retrospective records and prospectively planned matings, were classified into five mating types: (1) affected to affected, (2) homozygous normal sire to any dam, (3) heterozygous to heterozygous, (4) heterozygous sire to affected dam, and (5) affected sire to heterozygous dam. For all but the last category, results were in accord with mendelian expectations for autosomal-recessive inheritance. However, litters of mating type 5 had fewer affected pups (20/77) than expected. The observed segregation ratio for this mating type (0.26) …
Metabolic Labeling Of Rod Outer Segment Phospholipids In Miniature Poodles With Progressive Rod-Cone Degeneration (Prcd), Mary Wetzel, Christian Fahlman, Paul O'Brien, Gustavo Aguirre
Dec 1989
Metabolic Labeling Of Rod Outer Segment Phospholipids In Miniature Poodles With Progressive Rod-Cone Degeneration (Prcd), Mary Wetzel, Christian Fahlman, Paul O'Brien, Gustavo Aguirre
Gustavo D. Aguirre, VMD, PhD
The recessive genetic defect in miniature poodles which results in progressive rod-cone degeneration (prcd) has been investigated in an attempt to determine the biochemical abnormality involved. In the present study, the rod outer segments of young prcd affected miniature poodles and normal dogs have been compared with respect to the incorporation of intravitreally injected [3H]palmitic acid, [14C]linolenic acid, and [14C]docosahexaenoic acid into neutral lipids and phospholipids as well as [3H]palmitate and [14C]leucine into rhodopsin. In addition, 3 mm trephined punches of retinas were incubated with [ …
Frequency Of The Codon 807 Mutation In The Cgmp Phosphodiesterase Β-Subunit Gene In Irish Setters And Other Dog Breeds With Hereditary Retinal Degeneration, Gustavo D. Aguirre, V Baldwin, K M. Weeks, Gregory M. Acland, Kunal Ray
Dec 1989
Frequency Of The Codon 807 Mutation In The Cgmp Phosphodiesterase Β-Subunit Gene In Irish Setters And Other Dog Breeds With Hereditary Retinal Degeneration, Gustavo D. Aguirre, V Baldwin, K M. Weeks, Gregory M. Acland, Kunal Ray
Gustavo D. Aguirre, VMD, PhD
Rod-cone dysplasia 1 (rcd1) in Irish setters is caused by a nonsense mutation in the cGMP phosphodiesterase β-subunit gene (PDE6B). We examined the frequency of the mutant allele in the Irish setter population and determined if the defect is present in dogs of other breeds which are affected with other inherited photoreceptor diseases. Between 1994 and 1997, samples were obtained from 436 clinically normal Irish setters, a red wolf, and dogs from 23 different breeds. The mutation in codon 807 of PDE6B was detected in genomic DNA by heteroduplex analysis, allele-specific PCR, or restriction enzyme digestion. …
Retinal Pigment Epithelial Glycosaminoglycan Metabolism: Intracellular Versus Extracellular Pathways. In Vitro Studies In Normal And Diseased Cells, Gustavo D. Aguirre, Lawrence E. Stramm, M Haskins
Sep 1989
Retinal Pigment Epithelial Glycosaminoglycan Metabolism: Intracellular Versus Extracellular Pathways. In Vitro Studies In Normal And Diseased Cells, Gustavo D. Aguirre, Lawrence E. Stramm, M Haskins
Gustavo D. Aguirre, VMD, PhD
The synthesis and turnover of glycosaminoglycans (GAGs) in different fractions of cultured feline retinal pigment epithelium (RPE) were characterized. In one method of fractionation, trypsin was used to separate the extracellular components (referred to as trypsin-soluble glycocalyx) from the intracellular components. As a second method, the basal extracellular matrix (basal ECM) was separated from the rest of the GAGs (cell-associated GAGs) by extracting the cell layer with NH4OH. The incorporation of 35SO4 into cetylpyridinium chloride-precipitable GAGs in the cell-associated and the intracellular fractions increased throughout the labeling period, while in the trypsin-soluble glycocalyx and the basal ECM incorporation approached a …
S-Antigen In A Hereditary Visual Cell Disease. Immunocytochemical And Immunological Studies, Gustavo D. Aguirre, K Long, N Philp, I Gery
Oct 1988
S-Antigen In A Hereditary Visual Cell Disease. Immunocytochemical And Immunological Studies, Gustavo D. Aguirre, K Long, N Philp, I Gery
Gustavo D. Aguirre, VMD, PhD
S-antigen is a photoreceptor-specific and potentially autoantigenic protein. Using light microscopic immunocytochemistry, the localization of S-antigen was studied in the retinas of normal dogs and Irish setters affected with rod-cone dysplasia, a hereditary retinal degeneration characterized by abnormal cGMP metabolism and arrested outer segment differentiation. Normal and affected dogs were also tested for the presence of humoral and cellular immunity to S-antigen. S-antigen was present in both rods and cones during inner and outer segment differentiation, but there was an apparent loss of immunoreactivity in cones as the retina matured. The developmental appearance and localization of S-antigen in affected retinas …
