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Abstract

αB-crystallin is a member of a small family of thermal shock proteins that protects cells from stress. Because of lack of its expression in peripheral blood leukocytes, it was proposed as a molecular marker of circulating tumor cells in canine mammary gland tumors. The aim of the present study was to determine if αB-crystallin shows stability of expression, what is the requirement for this type of marker. It was also assessed whether there is co-expression of αB-crystallin with the basal marker, cytokeratin 17. For this purpose, samples of various types of canine mammary gland tumors of epithelial origin, were selected. Using RT-qPCR, we have found αB-crystallin and cytokeratin 17 co-expression in benign and malignant canine mammary gland tumors. It has been demonstrated that the expression of αB-crystallin in tested neoplastic samples is not stable in comparison to the control group. Furthermore αB-crystallin overor down- expression was associated witch the same cytokeratin 17 pattern. αB-crystallin can be a marker of circulating tumor cells in the bloodstream, but for cancers in which basal marker expression occurs and thus not universal for all cancers originating from the mammary gland tissue.

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Authors and Affiliations

M. Chmielewska-Krzesińska
A. Jakimiuk
K. Wąsowicz
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Abstract

Combined retrograde tracing and double-labelling immunofluorescence were used to investigate the distribution and chemical coding of neurons in testicular (TG) and aorticoerenal (ARG) ganglia supplying the urinary bladder trigone (UBT) in juvenile male pigs (n=4, 12 kg. of body weight). Retrograde fluorescent tracer Fast Blue (FB) was injected into the wall of the bladder trigone under pentobarbital anesthesia. After three weeks all the pigs were deeply anesthetized and transcardially perfused with 4% buffered paraformaldehyde. TG and ARG, were collected and processed for double-labelling immunofluorescence. The expression of tyrosine hydroxylase (TH) or dopamine beta-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), nitric oxide synthase (NOS) and vesicular acetylcholine transporter (VAChT) were investigated. The cryostat sections were examined with a Zeiss LSM 710 confocal microscope equipped with adequate filter blocks.
The TG and ARG were found to contain many FB-positive neurons projecting to the UBT (UBT-PN). The UBT-PN were distributed in both TG and ARG. The majority of them were found in the right ganglia, mostly in TG. Immunohistochemistry disclosed that the vast majority of UBT-PN were noradrenergic (TH- and/or DBH-positive). Many noradrenergic neurons contained also immunoreactivity to NPY, SOM or GAL. Most of the UBT-PN were supplied with VAChT-, or NOS- IR (immunoreactive) varicose nerve fibres.
This study has revealed a relatively large population of differently coded prevertebral neurons projecting to the porcine urinary bladder. As judged from their neurochemical organization these nerve cells constitute an important element of the complex neuro-endocrine system involved in the regulation of the porcine urogenital organ function.
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Bibliography

De Groat WC (1995) Mechanisms underlying the recovery of lower urinary tract function following spinal cord injury. Paraplegia 33: 493-505.
De Groat W, Yoshimura N (2015) Anatomy and physiology of the lower urinary tract. Handb Clin Neurol 130: 61-108.
De Groat WC, Griffiths D, Yoshimura N (2015) Neural control of the lower urinary tract. Compr Physiol 5: 327-396.
Lepiarczyk E, Bossowska A, Skowrońska A, Majewski M (2019) A study on preganglionic connections and possible viscerofugal projec-tions from urinary bladder intramural ganglia to the caudal mesenteric ganglion in the pig. J Anat 234: 263-273.
Pidsudko Z (2013) Immunohistochemical characteristics and distribution of neurons in the intramural ganglia supplying the urinary bladder in the male pig. Pol J Vet Sci 16: 629-638.
Pidsudko Z (2014) Immunohistochemical characteristics and distribution of neurons in the paravertebral, prevertebral and pelvic ganglia sup-plying the urinary bladder in the male pig. J Mol Neurosci 52: 56-70.
Pidsudko Z, Majewski M (2004) The distribution and chemical coding of porcine urinary bladder trigone-projecting neurons located in pre-vertebral ganglia other than IMG. Pol J Vet Sci 7 (Suppl): 97-99.
Pidsudko Z, Kaleczyc J, Majewski M, Lakomy M, Scheuermann DW, Timmermans JP (2001) Differences in the distribution and chemical coding between neurons in the inferior mesenteric ganglion supplying the colon and rectum in the pig. Cell Tissue Res 303: 147-158.
Pidsudko Z, Listowska Z, Franke-Radowiecka A, Klimczuk M, Załȩcki M, Kaleczyc J (2019) Distribution and chemical coding of urinary bladder apex-projecting neurons in aorticorenal and testicular ganglia of the male pig. Pol J Vet Sci 22: 427-430.
Russo D, Clavenzani P, Sorteni C, Bo Minelli L, Botti M, Gazza F, Panu R, Ragionieri L, Chiocchetti R (2013) Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone. J Comp Neurol 521: 342-366.
Yamanishi T, Chapple C, Yasuda K, and Chess-Williams R (2002) The role of M2 muscarinic receptor subtypes in mediating contraction of the pig bladder base after cyclic adenosine monophosphate elevation and/or selective M3 inactivation. J Urol 167: 397-401.
Yoshimura N, Chancellor MB (2003) Neurophysiology of lower urinary tract function and dysfunction. Rev Urol (Suppl 8): S3-S10.
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Authors and Affiliations

Z. Pidsudko
1
J. Godlewski
2
K. Wąsowicz
3

  1. Department of Animal Anatomy, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego 13, 10-719 Olsztyn, Poland
  2. Department of Human Histology and Embryology, School of Medicine, University of Warmia and Mazury in Olsztyn, Warszawska 30, 10-082 Olsztyn, Poland
  3. Department of Pathophysiology, Forensic Veterinary Medicine and Administration, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego 14, 10-719 Olsztyn, Poland

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