Canine Brain and Tissue Bank

2017.05.01.
Canine Brain and Tissue Bank

Founded: 2017
Principal Investigator: Dr. Kubinyi Enikő
Funding: European Research Council (ERC), Hungarian Academy of Sciences National Brain Program
Contact: eniko.kubinyi@ttk.elte.hu


The Canine Brain and Tissue Bank is open to national and international collaborations. Currently, we store the brains and other samples from 148 family dogs from various breeds, 5 wolves, and 19 jackals.

This table contains information about the available sample types and dog breeds. We have RNAlater stabilized samples only from donations where the post-mortem delay was <4 hours.

Tissue type / region Storage method Number of animals Breeds Other species Remark
Whole brain (right hemisphere) Simply frozen at -80°C as a whole (old sampling procedure) 60 Beagle(10), Bichon havanaise (2); Border Collie(3); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Yorkshire terrier(2); Mongrel (variable,20)   Quality of samples may be questionable in cases where pieces have already been cut out from the brain tissue. We keep track of freeze-thawing cycles.
Whole brain (right hemisphere) Sliced and parts frozen separately at -80°C (current method) 18 Beagle (2); Borzoi(1); Whippet(1); Mongrel(5); White Swiss Shepherd Dog (1); Vizsla (1); Gordon Setter (1); Groenendael (1); Collie (1); German Shepherd Dog (2);Labrador Retriever (1); Staffordshire Terrier (1)    
Whole brain (or left hemisphere) Whole hemisphere fixed in 4% formaldehyde, stored at 4°C 135 Beagle(13); Bichon bolognese(1); Bichon frise(1); Bichon havanaise(3); Bobtail(1); Border Collie(3); Borzoi(1); Boxer(5); Caucasian Shepherd Dog(1); Chihuanua(1); ; Dachshund(1); Dobermann(1); English Bulldog(2); English Cocker Spaniel (1); German Shepherd(2); Golden retriever(3); Gordon Setter(1); Labrador retriever(6); Leonberger (1); Miniature poodle (2); Mudi(1); Puli (2); Rottweiler(1); Small Münsterlander(1); Staffordshire Terrier (1); Vizsla(2); Westie (2); Whippet(1); Yorkshire terrier(5); Mongrel (variable, ~30) Canis aureus (19) and Canis lupus (1)*, please ask for details  
Brain / frontal cortex Whole brain simply frozen at -80°C (1th phase sampling) 60 Beagle(10), Bichon havanaise (2); Border Collie(3); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Yorkshire terrier(2); Mongrel (variable,20)    
Brain / frontal cortex Sliced and frozen at -80°C (current method) 18 Beagle (1); Beagle (2); Borzoi(1); Whippet(1); Mongrel(5); White Swiss Shepherd Dog (1); Vizsla (1); Gordon Setter (1); Groenendael (1); Collie (1); German Shepherd Dog (2);Labrador Retriever (1); Staffordshire Terrier (1)    
Brain / frontal cortex RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21) female Canis lupus (4), please ask for details Some samples have already been used up for research, please ask for details.
Brain / frontal cortex Fixed in 4 % formaldehyde as whole brain hemisphere 135 Beagle(13); Bichon bolognese(1); Bichon frise(1); Bichon havanaise(3); Bobtail(1); Border Collie(3); Borzoi(1); Boxer(5); Caucasian Shepherd Dog(1); Chihuanua(1); ; Dachshund(1); Dobermann(1); English Bulldog(2); English Cocker Spaniel (1); German Shepherd(2); Golden retriever(3); Gordon Setter(1); Labrador retriever(6); Leonberger (1); Miniature poodle (2); Mudi(1); Puli (2); Rottweiler(1); Small Münsterlander(1); Staffordshire Terrier (1); Vizsla(2); Westie (2); Whippet(1); Yorkshire terrier(5); Mongrel (variable, ~30)   Some samples have already been used up for research, please ask for details.
Brain / cerebellum Whole brain simply frozen at -80°C (1th phase sampling) 60 Beagle(10), Bichon havanaise (2); Border Collie(3); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Yorkshire terrier(2); Mongrel (variable,20)    
Brain / cerebellum Sliced and frozen at -80°C (current method) 18 Beagle (2); Borzoi(1); Whippet(1); Mongrel(5); White Swiss Shepherd Dog (1); Vizsla (1); Gordon Setter (1); Groenendael (1); Collie (1); German Shepherd Dog (2);Labrador Retriever (1); Staffordshire Terrier (1)    
Brain / cerebellum RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21) female Canis lupus (4), please ask for details Some samples have already been used up for research, please ask for details.
Brain / cerebellum Fixed in 4 % formaldehyde as whole brain hemisphere 135 Beagle(13); Bichon bolognese(1); Bichon frise(1); Bichon havanaise(3); Bobtail(1); Border Collie(3); Borzoi(1); Boxer(5); Caucasian Shepherd Dog(1); Chihuanua(1); ; Dachshund(1); Dobermann(1); English Bulldog(2); English Cocker Spaniel (1); German Shepherd(2); Golden retriever(3); Gordon Setter(1); Labrador retriever(6); Leonberger (1); Miniature poodle (2); Mudi(1); Puli (2); Rottweiler(1); Small Münsterlander(1); Staffordshire Terrier (1); Vizsla(2); Westie (2); Whippet(1); Yorkshire terrier(5); Mongrel (variable, ~30)   Some samples have already been used up for research, please ask for details.
Brain / brain stem Whole brain simply frozen at -80°C (1th phase sampling) 60 Beagle(10), Bichon havanaise (2); Border Collie(3); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Yorkshire terrier(2); Mongrel (variable,20)    
Brain / brain stem Sliced and frozen at -80°C (current method) 25 Beagle (2); Borzoi(1); Whippet(1); Mongrel(5); White Swiss Shepherd Dog (1); Vizsla (1); Gordon Setter (1); Groenendael (1); Collie (1); German Shepherd Dog (2);Labrador Retriever (1); Staffordshire Terrier (1)    
Brain / brain stem RNA later stabilized tissue piece
64
Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21) Canis lupus (4), please ask for details Some samples have already been used up for research, please ask for details.
Brain / brain stem Fixed in 4 % formaldehyde as whole brain hemisphere 135 Beagle(13); Bichon bolognese(1); Bichon frise(1); Bichon havanaise(3); Bobtail(1); Border Collie(3); Borzoi(1); Boxer(5); Caucasian Shepherd Dog(1); Chihuanua(1); ; Dachshund(1); Dobermann(1); English Bulldog(2); English Cocker Spaniel (1); German Shepherd(2); Golden retriever(3); Gordon Setter(1); Labrador retriever(6); Leonberger (1); Miniature poodle (2); Mudi(1); Puli (2); Rottweiler(1); Small Münsterlander(1); Staffordshire Terrier (1); Vizsla(2); Westie (2); Whippet(1); Yorkshire terrier(5); Mongrel (variable, ~30)   Some samples have already been used up for research, please ask for details.
Skin from head RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21)    
Skin from nose RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21)    
Musculus temporalis RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21)   Some samples have already been used up for research, please ask for details.
Ggl. trigeminale RNA later stabilized tissue piece 64 Beagle(11), Bichon havanaise (2); Border Collie(3); Borzoi(1); Boxer(1);Chihuanua(1); Dachshund(1);Dobermann(1); German Shepherd(1); Golden retriever(2); Gordon Setter(1); Vizsla(2); Labrador retriever(5);Miniature poodle (2); Puli (1); Small münsterlander(1); Whippet(1); Yorkshire terrier(2); Mongrel (variable,21)    
Ggl. distale RNA later stabilized tissue piece 50 Beagle (7); Border Collie (2); Bichon Havanaise (2); Boxer (1); Chihuahua (1); Dachshund(1); Dobermann(1); German Shepherd Dog (3); Golden Retriever (2); Gordon Setter (2); Labrador Retriever (4); Miniature poodle (2); Small Münsterlander (1); Vizsla (2); Yorkshire Terrier (2); Mongrel (12); White Swiss Shepherd Dog (1); Vizsla (1); Groenendael (1); Collie (1); Staffordshire Terrier (1)   Standard collection was discontinued from 2019.
Ggl. Cervicale (C4 or C5*) RNA later stabilized tissue piece 50 Beagle (7); Border Collie (2); Bichon Havanaise (2); Boxer (1); Chihuahua (1); Dachshund(1); Dobermann(1); German Shepherd Dog (3); Golden Retriever (2); Gordon Setter (2); Labrador Retriever (4); Miniature poodle (2); Small Münsterlander (1); Vizsla (2); Yorkshire Terrier (2); Mongrel (12); White Swiss Shepherd Dog (1); Vizsla (1); Groenendael (1); Collie (1); Staffordshire Terrier (1)   Standard collection was discontinued from 2019.
Gl. thyreoidea RNA later stabilized tissue piece 50 Beagle (7); Border Collie (2); Bichon Havanaise (2); Boxer (1); Chihuahua (1); Dachshund(1); Dobermann(1); German Shepherd Dog (3); Golden Retriever (2); Gordon Setter (2); Labrador Retriever (4); Miniature poodle (2); Small Münsterlander (1); Vizsla (2); Yorkshire Terrier (2); Mongrel (12); White Swiss Shepherd Dog (1); Vizsla (1); Groenendael (1); Collie (1); Staffordshire Terrier (1)   Standard collection was discontinued from 2019.
Ln. retropharyngeus medialis RNA later stabilized tissue piece 50 Beagle (7); Border Collie (2); Bichon Havanaise (2); Boxer (1); Chihuahua (1); Dachshund(1); Dobermann(1); German Shepherd Dog (3); Golden Retriever (2); Gordon Setter (2); Labrador Retriever (4); Miniature poodle (2); Small Münsterlander (1); Vizsla (2); Yorkshire Terrier (2); Mongrel (12); White Swiss Shepherd Dog (1); Vizsla (1); Groenendael (1); Collie (1); Staffordshire Terrier (1)   Standard collection was discontinued from 2019.
Liver RNA later stabilized tissue piece 18 Border Collie; Puli; Borzoi; Beagle; Vizsla; Wolf; Bull terrier; German shepherd dog (2);Labrador Retriever; Staffordshire Terrier; Mongrel (5)   Standard collection started from 2020.
Tumors RNA later stabilized tissue piece       Samples collected on ocassion, ask for more details!

Donating the bodies of euthanized pet dogs is a voluntary action based on the concordant decision of the dogs’ owners and the veterinarians who would perform the euthanasia. Both the owners and veterinarians have to fill out a Donation Document, which confirms the ownership of the dog and that it had valid vaccination against rabies before the CBTB can accept the actual donation.

In all cases when the CBTB gets informed about an expected donation, the transportation of the cadavers is arranged before the euthanasia takes place in order to minimalise the elapsed time between the death of the animal and the fixation of tissue samples. Therefore, we have been able to obtain molecular grade (stabilized in RNAlater, Thermo Fisher)  tissue samples  within four hours post mortem from 61 animals so far. These samples are being stored at -80 °C for the long term. After taking tissue pieces from each target area (see below) and putting them in RNAlater, we process the whole brain: the right hemispheres get stored directly at -80 °C and the left hemispheres get fixed in 10% formaldehyde solution and stored at +4 °C.

In other cases, where the post mortem delay exceeds this limit, only the brains of the animals are collected and get stored primarily for histological purposes. 

Tissue samples, which have been fixed in RNAlater within 4 hours post mortem, are generally available from several areas.
The locations from where we have collected RNAlater fixed tissue pieces from all animals (cases with <4 hours post mortem time) are the following: nose skin; skin from the head; temporal muscle; trigeminal ganglion (whole ganglion); prefrontal cortex (gyrus compositus anterior); cerebellum; brain stem.
We also collect and store liquor cerebrospinalis, when it is obtainable (e.g. the animals were not extensively dehydrated, in which cases the liquor has a very limited accessible volume).
Buccal swabs and hair are also taken from all animals.

Furthermore, tissue pieces from some animals (<4 hours port mortem) from the following locations are also available: thyroid gland; retropharyngeal medial lymph node; liver and heart.

Cases with > 4 hours post mortem delay

We have formaldehyde-fixed brains from several animals. We can provide information about these samples on demand.

Research and Perspectives

Our own research goals mainly focus on canine ageing. We are currently investigating gene expression differences on the mRNA level, both by specific (targeting previously described genes) and high-throughput (RNA sequencing) approaches.

Through the first years of establishing the CBTB sampling and sample processing protocols we have monitored the quality of the samples by RT-qPCR, comparing the stability of housekeeping genes (GAPDH, HMBS and HPRT1) between donated animals with different sampling times (within the 4 hours limit). As we haven’t detected a correlation between RNA quality and sampling time within this time frame, we plan to extend the time limit for collecting molecular-grade samples. This will allow us to assign more donated animals for RNA-based research goals.

Our protocol which can allow efficient stabilization and subsequent purification of proteins for various research goals is still under development. As the RNAlater fixed samples are primarily intended for RNA analyses and there seems to be a demand for them, we would like to collect and store extra samples from the same animals, which will be primarily intended for proteomics investigations.

Publications

  1. Kovács, T., Szinyákovics, J., Billes, V., Murányi, G., Varga, V.B., Bjelik, A., Légrádi, Á., Szabó, M., Sándor, S., Kubinyi, E. and Szekeres-Paracky, C., 2022. A conserved MTMR lipid phosphatase increasingly suppresses autophagy in brain neurons during aging. Scientific Reports, 12(1), p.21817. https://www.nature.com/articles/s41598-022-24843-w
  2. Sándor S, Urfer S, Kubinyi E. Toward establishing a worldwide net of canine biobanks. Aging (Albany NY), 14(6), 2436–2437. https://www.aging-us.com/article/203961/text
  3. Sándor, S., Jónás, D., Tátrai, K., Czeibert, K., Kubinyi, E. (2022) Poly(A) RNA sequencing reveals age-related differences in the prefrontal cortex of dogs. GeroScience, 44, 1269–1293. https://doi.org/10.1007/s11357-022-00533-3
  4. Urfer, S.R., Darvas, M., Czeibert, K., Sándor, S. Promislow, D. E. L., Creevy, K. E., Kubinyi*, E., Kaeberlein*, M. (2021) Canine Cognitive Dysfunction (CCD) scores correlate with amyloid beta 42 levels in dog brain tissue. GeroScience, 43:2379–2386. https://doi.org/10.1007/s11357-021-00422-1. *These authors jointly supervised this work.
  5. Sándor, S., Czeibert, K., Salamon, A., & Kubinyi, E. (2021). Man’s best friend in life and death: scientific perspectives and challenges of dog brain banking. GeroScience, 43:1653–1668. https://link.springer.com/article/10.1007/s11357-021-00373-7
  6. Sándor, S., Tátrai, K., Czeibert, K., Egyed, B., & Kubinyi, E. (2021). CDKN2A gene expression as a potential aging biomarker in dogs. Frontiers in Veterinary Science, 8, 348. https://www.frontiersin.org/articles/10.3389/fvets.2021.660435/full10.3389/fvets.2021.660435